CA2488682C - Gene differentially expressed in breast and bladder cancer and encoded polypeptides - Google Patents

Abstract

The present invention relates to a novel human gene that is differentially expressed in human carcinoma. More specifically, the present invention relates to a polynucleotide encoding a novel human polypeptide named C35 that is overexpressed in human breast and bladder carcinoma. This invention also relates to C35 polypeptide, in particular C35 peptide epitopes and C35 peptide epitope analogs, as well as vectors, host cells, antibodies directed to C35 polypeptides, and the recombinant methods for producing the same. The present invention further relates to diagnostic methods for detecting carcinomas, including human breast carcinomas. The present invention further relates to the formulation and use of the C35 gene and polypeptides, in particular C35 peptide epitopes and C35 peptide epitope analogs, in immunogenic compositions or vaccines, to induce antibody or cell-mediated immunity against target cells, such as tumor cells, that express the C35 gene. The invention further relates to screening methods for identifying agonists and antagonists of C35 activity.

Description

DEMANDES OU BREVETS VOLUMINEUX
LA PRESENTE PARTIE DE CETTE DEMANDE OU CE BREVETS
COMPREND PLUS D'UN TOME.

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GENE DIFFERENTIALLY EXPRESSED IN BREAST AND BLADDER
CANCER AND ENCODED POLYPEPTIDES
BACKGROUND OF THE INVENTION
Field of the Invention [0001] The present invention relates to a novel human gene that is differentially expressed in human breast and bladder carcinoma. More specifically, the present invention relates to a polynucleotide encoding a novel human polypeptide named C35. This invention also relates to C35 polypeptides, as well as vectors, host cells, antibodies directed to C35 polypeptides, and the recombinant methods for producing the same. The present invention further relates to diagnostic methods for detecting carcinomas, including human breast and bladder carcinomas. The present invention further relates to the formulation and use of the C35 gene and polypeptides in immunogenic compositions or vaccines, to induce antibody and cell-mediated immunity against target cells, such as tumor cells, that express the C35 gene. The invention further relates to screening methods for identifying agonists and antagonists of C35 activity.
Background Art [0002] Cancer afflicts approximately 1.2 million people in the United States each year. About 50% of these cancers are curable with surgery, radiation therapy, and chemotherapy. Despite significant technical advances in these three types of treatments, each year more than 500,000 people will die of cancer in the United States alone. (Jaffee, E. M., Ann. N. Y. Acad. Sci. 886:67-72 (1999)). Because most recurrences are at distant sites such as the liver, brain, bone, and lung, there is an urgent need for improved systemic therapies.

[0003] The goal of cancer treatment is to develop modalities that specifically target tumor cells, thereby avoiding unnecessary side effects to normal tissue.
Immunotherapy has the potential to provide an alternative systemic treatment for most types of cancer. The advantage of immunotherapy over radiation and chemotherapy is that it can act specifically against the tumor without causing normal tissue damage. One form of immunotherapy, vaccines, is particularly attractive because they can also provide for active immunization, which allows for amplification of the immune response. In addition, vaccines can generate a memory immune response.

[0004] The possibility that altered features of a tumor cell are recognized by the immune system as non-self and may induce protective immunity is the basis for attempts to develop cancer vaccines. Whether or not this is a viable strategy depends on how the features of a transformed cell are altered. Appreciation of the central role of mutation in tumor transformation gave rise to the hypothesis that tumor antigens arise as a result of random mutation in genetically unstable cells.
Although random mutations might prove immunogenic, it would be predicted that these would induce specific immunity unique for each tumor. This would be unfavorable for development of broadly effective tumor vaccines. An alternate hypothesis, however, is that a tumor antigen may arise as a result of systematic and reproducible tissue specific gene deregulation that is associated with the transformation process. This could give rise to qualitatively or quantitatively=
different expression of shared antigens in certain types of tumors that might be suitable targets for immunotherapy. Early results, demonstrating that the immunogenicity of some experimental tumors could be traced to random mutations (De Plaen, et al., Proc. Natl. Acad. Sci. USA 85: 2274-2278 (1988);
Srivastava, & Old, ImmunoL Today 9:78 (1989)), clearly supported the first hypothesis. There is, however, no a priori reason why random mutation and systematic gene deregulation could not both give rise to new immunogenic expression in tumors. Indeed, more recent studies in both experimental tumors (Sahasrabudhe et al., J. ImmunoL /51:6202-6310 (1993); Torigoe et al., J.
ImmunoL /47:3251 (1991)) and human melanoma (van Der Bruggen et al., Science 254:1643-1647(1991); Brichard et al. , J. Exp. Med. 178:489-495(1993);

Kawakami etal., Proc. NatL Acad. Sci. USA 91:3515-3519 (1994); Boel etal., Immunity 2:167-175 (1995); Van den Eynde et al., J. Exp. Med. 182: 689-698 (1995)) have clearly demonstrated expression of shared tumor antigens encoded by deregulated normal genes. The identification of MAGE-1 and other antigens common to different human melanoma holds great promise for the future development of multiple tumor vaccines.

[0005] In spite of the progress in melanoma, very few shared antigens recognized by cytotoxic T cells have not been described for other human tumors. The major challenge is technological. The most widespread and to date most successful approach to identify immunogenic molecules uniquely expressed in tumor cells is to screen a cDNA library with tumor-specific CTLs (cytotoxic T
lymphocytes).
Application of this strategy has led to identification of several gene families expressed predominantly in human melanoma. Two major limitations of this approach, however, are that (1) screening requires labor intensive transfection of numerous small pools of recombinant DNA into separate target populations, which themselves often need to be modified to express one or more MHC
molecules required for antigen presentation, in order to assay T cell stimulation by a minor component of some pool; and (2) with the possible exception of renal cell carcinoma, tumor-specific CTLs have been very difficult to isolate from either tumor infiltrating lymphocytes (TIL) or PBL of patients with other types of tumors, especially the epithelial cell carcinomas that comprise greater than 80% of human tumors. It appears that there may be tissue specific properties that result in tumor-specific CTLs being sequestered in melanoma.

[0006] Direct immunization with tumor-specific gene products may be essential to elicit an immune response against some shared tumor antigens. It has been argued that, if a tumor expressed strong antigens, it should have been eradicated prior to clinical manifestation. Perhaps then, tumors express only weak antigens.
Immunologists have long been interested in the issue of what makes an antigen weak or strong. There have been two major hypotheses. Weak antigens may be poorly processed and fail to be presented effectively to T cells.
Alternatively, the number of T cells in the organism with appropriate specificity might be inadequate for a vigorous response (a so-called "hole in the repertoire").
Elucidation of the complex cellular process whereby antigenic peptides associate with MHC molecules for transport to the cell surface and presentation to T
cells has been one of the triumphs of modern immunology. These experiments have clearly established that failure of presentation due to processing defects or competition from other peptides could render a particular peptide less immunogenic. In contrast, it has, for technical reasons, been more difficult to establish that the frequency of clonal representation in the T cell repertoire is an important mechanism of low responsiveness. Recent studies demonstrating that the relationship between immunodominant and cryptic peptides of a protein antigen change in T cell receptor transgenic mice suggest, however, that the relative frequency of peptide-specific T cells can, indeed, be a determining factor in whether a particular peptide is cryptic or dominant in a T cell response.
This has encouraging implications for development of vaccines. With present day methods, it would be a complex and difficult undertaking to modify the way in which antigenic peptides of a tumor are processed and presented to T cells.
The relative frequency of a specific T cell population can, however, be directly and effectively increased by prior vaccination. This could, therefore, be the key manipulation required to render an otherwise cryptic response immunoprotective.
These considerations of cryptic or sub-dominant antigens have special relevance in relation to possible immune evasion by tumors through tolerance induction.
Evidence has been presented to suggest that tumor-specific T cells in the tumor-bearing host are anergic, possibly as a result of antigen presentation on non-professional APC (Morgan, D.J. et al., J. Immunol. /63:723-27 (1999);
Sotomayor, E.M. etal., Proc. Natl. Acad. Sci. U.S.A. 96:11476-81 (1999); Lee, P.P. et al., Nature Medicine 5:677-85 (1999)). Prior tolerization of T cells specific for immunodominant antigens of a tumor may, therefore, account for the difficulty in developing successful strategies for immunotherapy of cancer.
These observations suggest that T cells specific for immunodominant tumor antigens are less likely to be effective for immunotherapy of established tumors because they are most likely to have been tolerized. It may, therefore, be that T cells specific for sub-dominant antigens or T cells that are initially present at a lower frequency would prove more effective because they have escaped the tolerizing influence of a growing tumor.

[0007] Another major concern for the development of broadly effective human vaccines is the extreme polymorphism of HLA class I molecules. Class I
MHC:cellular peptide complexes are the target antigens for specific CD8+ CTLs.

The cellular peptides, derived by degradation of endogenously synthesized proteins, are translocated into a pre-Golgi compartment where they bind to class I MEC molecules for transport to the cell surface. The CD8 molecule contributes to the avidity of the interaction between T cell and target by binding to the a3 domain of the class I heavy chain. Since all endogenous proteins turn over, peptides derived from any cytoplasmic or nuclear protein may bind to an MEIC
molecule and be transported for presentation at the cell surface. This allows T
cells to survey a much larger representation of cellular proteins than antibodies which are restricted to recognize conformational determinants of only those proteins that are either secreted or integrated at the cell membrane.

[0008] The T cell receptor antigen binding site interacts with determinants of both the peptide and the surrounding MHC. T cell specificity must, therefore, be defined in terms of an MHC:peptide complex. The specificity of peptide binding to MHC molecules is very broad and of relatively low affinity in comparison to the antigen binding sites of specific antibodies. Class I-bound peptides are generally 8-10 residues in length and accommodate amino acid side chains of restricted diversity at certain key positions that match pockets in the MHC
peptide binding site. These key features ofpeptides that bind to a particular MHC
molecule constitute a peptide binding motif.

[0009] Hence, there exists a need for methods to facilitate the induction and isolation of T cells specific for human tumors, cancers and infected cells and for methods to efficiently select the genes that encode the major target antigens recognized by these T cells in the proper MHC-context.

-o-BRIEF SUMMARY OF THE INVENTION
An object of the present invention is to provide a gene differentially expressed in breast and bladder cancer and encoded polypeptides.
The present invention relates to a novel polynucleotide, C35, that is differentially expressed in human breast and bladder carcinoma, and to the encoded polypeptide of C35. Moreover, the present invention relates to vectors, host cells, antibodies, and recombinant methods for producing C35 polypeptides and polynucleotides. The present invention further relates to the formulation and use of C35 polypeptides, in particular C35 peptide epitopes and C35 peptide epitope analogs, and polynucleotides in immunogenic compositions to induce antibodies and cell-mediated immunity against target cells, such as tumor cells, that express the C35 gene products. Also provided are diagnostic methods for detecting disorders relating to the C35 genes and polypeptides, including use as a prognostic marker for carcinomas, such as human breast carcinoma, and therapeutic methods for treating such disorders. The invention further relates to screening methods for identifying binding partners of C35.
Thus, in one embodiment, the invention relates to an isolated polypeptide comprising a peptide comprising two or more C35 peptide epitopes, wherein said peptide is selected from the group consisting of: amino acids T101 to V113 of SEQ BD NO:2, E100 to V113 of SEQ ID NO:2, G99 to V113 of SEQ ID NO:2, 193 to V113 of SEQ ID NO:2, D88 to V113 of SEQ ID NO:2, P84 to V113 of SEQ ID NO:2, K77 to V113 of SEQ ID NO:2, Q72 to V113 of SEQ ED NO:2, F65 to V113 of SEQ ID NO:2, and L59 to V113 of SEQ ID NO:2, and wherein said isolated polypeptide is not SEQ ID NO: 2, SEQ lID NO: 153, SEQ El) NO:
155, or amino acids E100 to R109 of SEQ ID NO:2.
In another embodiment, the invention relates to an isolated polypeptide comprising at least one C35 peptide epitope analog, wherein said C35 peptide epitope analog is selected from the group consisting of: for the peptide epitope G22 to C30 of SEQ ED NO:2 and FIG. 1B, the analogs with either alanine or glycine substituted for cysteine at the ninth amino acid residue; for the peptide epitope 125 to C33 of SEQ ID NO:2 and FIG. 1B, the analogs with either alanine or glycine substituted for the cysteine at the sixth amino acid residue and/or the ninth amino acid residue; for the peptide epitope K77 to Y85 of SEQ ID NO: 2 and FIG. 1B, the analog with valine substituted for tyrosine at the ninth amino acid residue; for the peptide epitope K104 to C112 of SEQ ID NO:2 and FIG. 1B, the analogs with alanine, glycine or leucine substituted for cysteine at the ninth amino acid residue; for the peptide epitope K104 to V113 of SEQ ID NO:2 and FIG. 1B, the analogs with alanine, serine, glycine or leucine substituted for cysteine at the ninth amino acid residue; for the peptide epitope 1105 to V113 of SEQ ID NO:2 and FIG. 1B, the analogs with either leucine or methionine substituted for threonine at the second amino acid residue and/or alanine, serine or glycine substituted for cysteine at the eighth amino acid residue; and for the peptide epitope N107 to L115 of SEQ ID NO:2 and FIG. 1B, the analog with either alanine or glycine substituted for cysteine at the sixth amino acid residue.
[0013] Preferably the isolated polypeptide of the invention is not more than 100 amino acids in length, alternatively not more that 95 amino acids in length, alternatively not more than 90 amino acids in length, alternatively not more than 85 amino acids in length, alternatively not more than 80 amino acids in length, alternatively not more than 75 amino acids in length, alternatively not more than 70 amino acids in length, alternatively not more than 65 amino acids in length, alternatively not more than 60 amino acids in length, alternatively not more than 55 amino acids in length, alternatively not more than 50 amino acids in length, alternatively not more than 45 amino acids in length, alternatively not more than 40 amino acids in length, or alternatively not more than 35 amino acids in length.
[0014] In another embodiment, the invention relates to a fusion protein comprising an isolated peptide comprising two or more C35 peptide epitopes, wherein said isolated peptide is selected from the group consisting of: amino acids T101 to V113 of SEQ ID NO:2, E100 to V113 of SEQ ID NO:2, G99 to V113 of SEQ ID NO:2, 193 to V113 of SEQ ID NO:2, D88 to V113 of SEQ ID
NO:2, P84 to V113 of SEQ ID NO:2, K77 to V113 of SEQ ID NO:2, Q72 to V113 of SEQ ID NO:2, F65 to V113 of SEQ ID NO:2, and L59 to V113 of SEQ

ID NO:2. In a preferred embodiment, the fusion protein is a homopolymer of said isolated peptide. In another preferred embodiment, the fusion protein is a heteropolymer of said isolated polypeptides. In yet another embodiment, the fusion protein is fused to a T helper peptide. In still another embodiment, the fusion protein is fused to a carrier. In another embodiment, the fusion protein is linked to a lipid.
[0015] In another embodiment, the invention relates to an isolated polypeptide consisting of two or more C35 peptide epitopes, wherein said isolated polypeptide is selected from the group consisting of: amino acids T101 to V113 of SEQ ID
NO:2, E100 to V113 of SEQ ID NO:2, G99 to V113 of SEQ ID NO:2, 193 to V113 of SEQ ID NO:2, D88 to V113 of SEQ ID NO:2, P84 to V113 of SEQ ID
NO:2, K77 to V113 of SEQ ID NO:2, Q72 to V113 of SEQ ID NO:2, F65 to V113 of SEQ ID NO:2, and L59 to V113 of SEQ ID NO:2, and wherein said isolated polypeptide is not SEQ ID NO: 2, SEQ ID NO: 153, SEQ ID NO: 155, or amino acids E100 to R109 of SEQ ID NO:2.
[0016] In another embodiment, the invention relates to an isolated polypeptide comprising a peptide comprising at least one C35 peptide epitope analog, wherein said peptide is selected from the group consisting of the analog of peptide to V113 of SEQ ID NO:2 having either alanine or glycine substituted for the cysteine at the twelfth residue, the analog of peptide El 00 to V113 of SEQ ID

NO:2 having either alanine or glycine substituted for the cysteine at the thirteenth residue, the analog ofpeptide G99 to V113 of SEQ ID NO:2 having either alanine or glycine substituted for cysteine at the fourteenth residue, the analog of peptide 193 to V113 of SEQ ID NO:2 having either alanine or glycine substituted for the cysteine at the twentieth residue, the analog of peptide D88 to V113 of SEQ ID

NO:2 having either alanine or glycine substituted for the cysteine at the twenty-fifth residue, the analog of peptide P84 to V113 of SEQ ID NO:2 having either alanine or glycine substituted for the cysteine at the twenty-ninth residue, the analog of peptide K77 to V113 of SEQ ID NO:2 having either alanine or glycine substituted for the cysteine at the thirty-sixth residue, the analog of peptide Q72 to V113 of SEQ ID NO:2 having either alanine or glycine substituted for the cysteine at the forty-first residue, the analog of peptide F65 to V113 of SEQ
ID
NO:2 having either alanine or glycine substituted for the cysteine at the forty-eighth residue, and the analog of peptide L59 to V113 of SEQ ID NO:2 having either alanine or glycine substituted for the cysteine at the fifty-fourth residue.
[0017] In another embodiment, the invention relates to a fusion protein comprising a peptide comprising at least one C35 peptide epitope analog, wherein said peptide is selected from the group consisting of: for the peptide epitope to C30 of SEQ ID NO:2 and FIG. 1B, the analogs with either alanine or glycine substituted for cysteine at the ninth amino acid residue; for the peptide epitope 125 to C33 of SEQ ID NO:2 and FIG. 1B, the analogs with either alanine or glycine substituted for the cysteine at the sixth amino acid residue and/or the ninth amino acid residue; for the peptide epitope K77 to Y85 of SEQ ID NO: 2 and FIG. 1B, the analog with valine substituted for tyrosine at the ninth amino acid residue; for the peptide epitope K104 to C112 of SEQ ED NO:2 and FIG. 1B, the analogs with alanine, glycine or leucine substituted for cysteine at the ninth amino acid residue; for the peptide epitope K104 to V113 of SEQ ID NO:2 and FIG. 1B, the analogs with alanine, glycine, serine or leucine substituted for cysteine at the ninth amino acid residue; for the peptide epitope 1105 to V113 of SEQ ID NO:2 and FIG. 1B, the analogs with either leucine, serine or methionine substituted for threonine at the second amino acid residue and/or alanine or glycine substituted for cysteine at the eighth amino acid residue; and for the peptide epitope N107 to V113 of SEQ ID NO:2 and FIG. 1B, the analog with either alanine or glycine substituted for cysteine at the sixth amino acid residue, the analog of peptide T101 to V113 of SEQ ID NO:2 having either alanine or glycine substituted for the cysteine at the twelfth residue, the analog of peptide E100 to V113 of SEQ ID NO:2 having either alanine or glycine substituted for cysteine at the thirteenth residue, the analog of peptide G99 to V113 of SEQ
ID
NO:2 having either alanine or glycine substituted for cysteine at the fourteenth residue, the analog of peptide 193 to V113 of SEQ ID NO:2 having either alanine or glycine substituted for the cysteine at the twentieth residue, the analog of peptide D88 to V113 of SEQ ID NO:2 having either alanine or glycine substituted for the cysteine at the twenty-fifth residue, the analog of peptide P84 to V113 of SEQ ID NO:2 having either alanine or glycine substituted for the cysteine at the twenty-ninth residue, the analog ofpeptide K77 to V113 of SEQ ID NO:2 having either alanine or glycine substituted for the cysteine at the thirty-sixth residue, the analog of peptide Q72 to V113 of SEQ ID NO:2 having either alanine or glycine substituted for the cysteine at the forty-first residue, the analog of peptide F65 to V113 of SEQ ID NO:2 having either alanine or glycine substituted for the cysteine at the forty-eighth residue, and the analog of peptide L59 to V113 of SEQ D NO:2 having either alanine or glycine substituted for the cysteine at the fifty-fourth residue. In a preferred embodiment, the fusion protein comprises a homopolymer of said peptide comprising at least one C35 peptide epitope analog.
In another preferred embodiment, the fusion protein comprises a heteropolymer of said peptide comprising at least one C35 peptide epitope analog.
In accordance with another aspect of the present invention, there is provided an isolated peptide comprising at least one C35 peptide epitope analog, wherein said peptide epitope analog is K104 to V113 of SEQ ID NO:2 and FIG. 1B, wherein the cysteine at the ninth amino acid residue is cysteinylated.
In accordance with another aspect of the present invention, there is provided an isolated peptide comprising at least one C35 peptide epitope analog, wherein said peptide epitope analog is 1105 to V113 of SEQ ID NO:2 and FIG. 1B, wherein the cysteine at the eighth amino acid residue is cysteinylated.

10a In another embodiment, the invention relates to a composition comprising an isolated polypeptide or fusion protein of the invention and a pharmaceutically acceptable carrier.
BRIEF DESCRIPTION OF THE FIGURES
FIGS. lA and 1B. FIG. lA shows the DNA coding sequence (SEQ ID
NO:1) of C35. The sequence immediately upstream of the predicted ATG start codon is shown in lower case and conforms to the expected features described by Kozak, M., J. Biol. Chem. 266(30):19867-19870 (1991). FIG. 1B shows the deduced amino acid sequence (SEQ ID NO:2) of C35.
FIGS. 2A-2C. FIG. 2A: C35 is overexpressed in Breast tumor cell lines.
Upper Panel: 300ng of poly-A RNA from 3 week old human thymus, normal breast epithelial cell line Hi 6N2 from patient 21, and 4 breast tumor cell lines derived one year apart from primary or metastatic nodules of the same patient 21;

21NT, 21PT 21MT1, and 21MT2, was resolved on a 1% agarose/formaldehyde gel and -transferred to a GeneScreen membrane. This blot was hybridized with a32P labeled C35 probe. Hybridization was detected by exposing the blot to film for 15 hours. Lower Panel: To quantitate RNA loading, the same blot was stripped and re-hybridized with a 32P labeled probe for Glyceraldehyde-3 Phosphate Dehydrogenase (GAPDH). For each sample the C35 signal was normalized to the GAPDH signal. The numbers represent the fold expression of C35 in each sample relative to H16N2. FIG. 2B: C35 is expressed at low levels in normal tissues. A Blot containing 1 microgram of poly-A RNA from each of the indicated adult normal tissues (Clontech) was hybridized with a 3213 labeled C35 probe. Hybridization was detected by exposing the blot to film for 15 hours (upper panel), or 96 hours (lower panel). FIG. 2C. C35 is overexpressed in primary Breast tumors. A blot containing 2 micrograms of poly-A RNA from 3 primary infiltrating ductal mammary carcinoma, Ti, T2, T3 and 1 normal breast epithelium, N (Invitrogen) was hybridized with a 32P labeled C35 probe. To normalize loading a 32P labeled beta-Actin probe was included in the hybridization mix. Hybridization was detected by exposing the blot to film for 6 hours. The numbers represent the fold expression of C35 in each sample relative to normal breast epithelium.
[0021] FIG. 3. Expression of C35 in Breast Tumor Cell Lines. C35 is overexpressed in different breast tumor cell lines. Upper Panel: 300ng of poly-A
RNA from BT474 (ATCC HYB-20, mammary ductal carcinoma), SKBR3 (ATCC HTB-30, mammary adenocarcinoma), T47D (ATCC HTB-133, mammary ductal carcinoma), normal breast epithelial cell line H16N2 from patient 21, and 21-NT breast tumor cell line derived from primary tumor nodule of the same patient 21 was resolved on a 1% agarose/formaldehyde gel and transferred to a GeneScreen membrane. This blot was hybridized with a 32P
labeled C35 probe. Hybridization was detected by exposing the blot to film for 15 hours. Lower Panel: To quantitate RNA loading, the same blot was stripped and re-hybridized with a 32P labeled probe for beta-actin. For each sample the C35 signal was normalized to the actin signal. The numbers represent the fold expression of C35 in each sample relative to H16N2.
[0022] FIGS. 4A-4C. Surface Expression of C35 Protein Detected by Flow Cytometry. 1 x 105 breast tumor cells were stained with 3.5 microliters of antiserum raised in BALB/c mice against Line 1 mouse tumor cells transduced with retrovirus encoding human C35 or control, pre-bleed BALB/c serum. After a 30 minute incubation, cells were washed twice with staining buffer (PAB) and incubated with FITC-goat anti-mouse IgG (1 ug/sample) for 30 minutes. Samples were washed and analyzed on an EPICS Elite flow cytometer. FIG. 4A: 21NT.
FIG. 4B: SKBR3. FIG. 4C: MDA-MB-231. These three breast tumor lines were selected to represent tumor cells that express high, intermediate and low levels of C35 RNA on Northern blots (see FIG. 3). Abbreviations: nms, ns;
normal mouse serum; C35; C35 immune serum.
[0023] FIGS. 5A and 5B. CML Selected Recombinant Vaccinia cDNA Clones Stimulate Tumor Specific CTL. FIG. 5A: CML Selected vaccinia clones were assayed for the ability, following infection of B/C.N, to stimulate tumor specific CTL to secrete interferon gamma. The amount of cytokine was measured by ELISA, and is represented as 0D490 (14). An 0D490 of 1.4 is approximately equal to 4 ng/ml of IFNg, and an 0D490 of 0.65 is approximately equal to 1 ng/ml of IENg. FIG. 5B: CML selected clones sensitize host cells to lysis by tumor specific CTL. Monolayers of B/C.N in wells of a 6 well plate were infected with moi=1 of the indicated vaccinia virus clones. After 14 hours of infection the infected cells were harvested and along with the indicated control targets labeled with51Cr. Target cells were incubated with the indicated ratios of tumor specific Cytotoxic T Lymphocytes for 4 hours at 37 C and percentage specific lysis was determined (15). This experiment was repeated at least three times with similar results.
[0024] FIGS. 6A and 6B. The Tumor Antigen Is Encoded by a Ribosomal Protein L3 Gene. Sequence of 112.16 and rpL3 from amino acid position 45 to 56. FIG. 6A: The amino acid (in single letter code) and nucleotide sequence of cDNA clone rpL3 (GenBank Accession no. Y00225). FIG. 6B: A single nucleotide substitution at C170T of the H2.16 tumor cDNA is the only sequence change relative to the published L3 ribosomal allele. This substitution results in a T54I amino acid substitution in the protein.
[0025] FIGS. 7A and 7B. Identification of the Peptide Epitope Recognized by the Tumor Specific CTL. FIG. 7A: CML assay to identify the peptide recognized by tumor specific CTL. Target cells were labeled with 51Cr (1 5) . During the 51Cr incubation samples of B/C.N cells were incubated with 1 RM peptide L348_56(I54), 100 i.tM L348_56(T54) or 100 M peptide L345_54(I54). Target cells were incubated with the indicated ratios of tumor specific Cytotoxic T Lymphocytes for 4 hours at 37 C and percentage specific lysis was determined. This experiment was repeated at least three times with similar results. FIG. 7B: Titration of peptide L348_56 (154). Target cells were labeled with 51Cr. During the 51Cr incubation samples o f B/C.N cells were incubated either with no peptide addition (D) or with the indicated concentrations (1 uM, 1 OnM, 1nM) of L348_56(154) (II), BCA 39 cells were included as a positive control (A). Target cells were incubated with the indicated ratios of Tumor Specific Cytotoxic T Lymphocytes for 4 hours at 37 C

and percentage specific lysis was determined. The experiment was repeated twice with similar results.
[0026] FIGS. 8A to 8C. Analysis of L3 Expressed by Each Cell Line. FIG.
8A:
Sau3AI map of published rpL3 and H2.16. Shown above is the Sau3AI
restriction map for the published ribosomal protein L3 gene (Top), and for H2.16 (Bottom). Digestion ofcDNA for the published L3 sequence generates fragments of 200, 355, 348, 289, and 84bp. The pattern for H2.16 is identical except for an extra Sau3A1 site at position 168 caused by the C170T. This results in a 168bp digestion product in place of the 200bp fragment. FIG. 8B: The BCA tumors express both L3 alleles. RT-PCR products generated from each cell line or from vH2.16 were generated using L3 specific primers and then digested with Sau3A1, and resolved on a 3% agarose gel for 2 hours at 80 volts. FIG. 8C: The Immunogenic L3 allele is expressed at greatly reduced levels in B/C.N, BCB 13, and Thymus. L3 specific RT-PCR products from each indicated sample were generated using a 321) end labeled 5 prime PCR primer. No PCR product was observed when RNA for each sample was used as template for PCR without cDNA synthesis, indicating that no sample was contaminated with genomic DNA. The PCR products were gel purified to ensure purity, digested with Sau3AI, and resolved on a 3% agarose gel for 15 hours at 60 volts. No PCR
product was observed in a control PCR sample that had no template added to it.

This result has been reproduced a total of 3 times.
[0027] FIGS. 9A to 9C. Immunization with iL3 is Immunoprotective. FIG. 9A:
Immunization with 112.16 induces tumor specific CTL. Balb/c mice (2/group) were immunized by subcutaneous injection with 5X106 pfu of vI12.16, or control vector v7.5/tk. Seven days later splenocytes were harvested and restimulated with peptide L348_56(I54) (26). Five days following the second restimulation the lymphocytes were tested in a chromium release assay as described in Figure 11.

The L348_56(T154) peptide was used at a 1 micromolar concentration, and the L348_ 56(T54) peptide was used at a 100 micromolar concentration. Similar results were obtained when the immunization experiment was repeated. FIGS. 9B and 9C:
Female Balb/cByJ mice were immunized as indicated (27). The mice were challenged by SC injection with 200,000 viable BCA 34 tumor cells into the abdominal wall. Data is from day 35 post challenge. These data are representative of 4 independent experiments.
[0028] FIGS. 10A and 10B. FIG. 10A: C35 coding sequence with translation;
5' and 3' untranslated regions are shown in lowercase letters. The predicted prenylation site, CVIL, at the 3' terminus is boxed. FIG. 10B: Genomic alignment of C35 gene on chromosome 17.
[0029] FIGS. 11A and 11B. C35 Expression in Breast Carcinoma. C35 was labeled with 32P in a random priming reaction and hybridized to Northern blots at 106 cpm/ml. Each blot was stripped and re-probed with GAPDH or Beta-actin to normalize mRNA loads. The numbers indicate densitometry ratios normalized against GAPDH/Beta-actin. A value of 1 has been assigned to normal cell line Hi 6N2, and all values are relative to the level of expression in the normal cell line. FIG. 11A: C35 expression in breast epithelial cell lines. FIG. 11B: C35 expression in primary breast tissue/tumors. 300 ng mRNA was electrophoresed on 0.8% alkaline agarose gels, then blotted to Genescreen Plus, except leftmost panel of B loaded with 1 ,ug mRNA from 3 primary tumors and 1 normal tissue control (Real Tumor Blots, Invitrogen). Similar exposures are shown for all blots.
[0030] FIG. 12. C35 Expression in Bladder Carcinoma. C35 was labeled with 32P in a random priming reaction and hybridized to a Northern blot of tumor and normal RNA at 106 cpm/ml. The blot was stripped and re-probed with Beta-actin to normalize mRNA loads. The numbers indicate densitometry ratios normalized against Beta-actin. Values are relative to the level of expression in the normal bladder samples. 300 ng mRNA was electrophoresed on 0.8% alkaline agarose gels, then blotted to Genescreen Plus.
[0031] FIGS. 13A and 13B. FACS Analysis with Anti-C35 Antibodies. FIG.
13A: Breast cell lines were stained with (top panel) sera from mice immunized with Line 1 cells infected with C35 recombinant retrovirus, and (bottom panel) 2C3 purified monoclonal antibody or isotype control. FIG. 13B: Bladder cell lines stained with 2C3 purified monoclonal antibody or isotype control.
[0032] FIGS. 14A and 14B. Inhibition of Tumor Growth in Presence of 2C3 Antibody. 21NT breast tumor cells (FIG. 14A) or H16N2 normal breast epithelial cells (FIG. 14B) were incubated with the indicated concentrations of 2C3 anti-C35 monoclonal antibody or a non-specific isotype control antibody.
Cell growth was measured by XTT assay following 72 hour incubation in the presence or absence of antibodies.
[0033] FIGS. 15A and 15B. CTL stimulated with C35 expressing dendritic cells specifically lyse C35+ Breast (21NT) and Bladder (ppT11A3) tumor cell lines, with minimal activity against normal breast (MEC), immortalized non-tumorigenic breast (Hi 6N2) and bladder (SV-HUC) cell lines, or an NK
sensitive cell line (K562). FIG. 15A: T cell line 4 was generated from normal human PBL. FIG. 15B: T cell clone 10G3 was selected from line 4 for C35-specific activity. Target cell lines MEC, ppT11A3 and SV-HUC are naturally HLA-A2 positive. Target cell lines 21NT and H16N2 were transected with HLA-A2 to provide a required MHC restriction element.
[0034] FIGS. 16A and 16B. Cytokine Release from T Cell Clone 10G3 upon Stimulation with Targets. FIG. 16A: IFN-gamma secretion. FIG. 16B: TNF-alpha secretion. Breast and bladder target cell lines were distinguished by the presence or absence of expression of HLA-A2 and C35 tumor antigen, an amino terminal 50 amino acid fragment of C35 (C35-50aa), or the irrelevant mouse L3 ribosomal protein. Each marker was either endogenously expressed or introduced by transfection of an HLA-A2.1 construct (pSV2.A2), or by infection with a vaccinia recombinant of C35 (w.C35, vv.C35-50aa), L3 (vv.L3), or HLA-A2 (vv.A2) [0035] FIGS. 17A and 17B. Effect of anti-CD40 ligand antibody (anti-CD154) in blocking the reactivity of murine T cells to specific transplantation antigens.
DBAJ2 (H-2d) mice were immunized with 107 C57B1/6 (H-21') spleen cells intraperitoneally and, in addition, were injected with either saline or 0.5 mg monoclonal anti-CD40 ligand antibody (MR1, anti-CD154, Pharmingen 09021D) administered both at the time of immunization and two days later. On day 10 following immunization, spleen cells from these mice were removed and stimulated in vitro with either C57B1/6 or control allogeneic C3H (H-2k) spleen cells that had been irradiated (20 Gy). After 5 days of in vitro stimulation, C57B1/6 and C3H specific cytolytic responses were assayed at various effector:target ratios by 51Cr release assay from specific labeled targets, in this case, either C3H or C57B1/6 dendritic cells pulsed with syngeneic spleen cell lysates. Significant cytotoxicity was induced against the control C3H
alloantigens in both saline and anti-CD154 treated mice (FIG. 17A) whereas a cytotoxic response to C57B1/6 was induced in the saline treated mice but not the anti-CD154 treated mice (FIG. 17B).

[0036] FIG. 18. GM-CSF Production by Line 4 After Stimulation with Native 21NT-A2 Tumor,1116N2-A2 Pulsed with Different C35 Peptides, or H16N2-A2 Infected with C35 Recombinant Vaccinia Virus. T cell line 4 was generated by stimulating normal donor T cells for 12 days each with autologous dendritic cells (DC) and then autologous monocytes infected with C35 recombinant vaccinia.virus. Weekly stimulation was continued with allo PBL and the 21NT
tumor transfected with HLA-A2/Kb (21NT-A2). For the experiment depicted here, the T cells were restimulated in vitro at 106 T cells per well with 5 x irradiated (2500 rads) H16N2-A2/Kb pulsed with lug/ml of C35 peptides 9-17, 77-85, 104-112, or 105-113 and 105 irradiated allo PBL per well with IL2 (20U/m1) and IL-7 (1 Ong/m1) in AIM-V/5% human AB serum. After two (2) rounds of stimulation for 7 days, T cells were tested for induction of GM-CSF
secretion following incubation with different stimulators pulsed or not pulsed with lug/ml of peptide or infected with vvC35 or vvWT at MOI = 1. T cells (5000) were incubated with 25000 of the various stimulator cells overnight in AIM-V/5% human AB serum in triplicate.
[0037] FIG. 19. C35-Specific ELISA of Hybridoma Supernatants. Results of a representative ELISA experiment involving hybridoma clones with demonstrated specificity for C35.
[0038] FIG. 20. Western Blot with C35-Specific Antibodies. Western Blot Immunodetection was performed with supernatant from selected hybridoma clones. Antibodies from 4 hybridomas (1B3, 1F2, 3E10, 11B10) reacted specifically with hC35 protein in this assay. Results for antibodies 1B3, 1F2, and 3E10 are shown. =
[0039] FIG. 21. Immunohistochemistry with 1F2 Antibody. Monoclonal antibody 1F2 was shown to have utility for immunohistochemical staining of primary breast tumor sections. This Figure demonstrates that monoclonal antibody 1F2 can detect high levels of endogenous C35 expression in human breast tumors, with little or no staining of normal breast tissue.
Specifically, this Figure shows strong staining of a section of invasive breast adenocarcinoma from patient 01A6, while normal breast tissue from the same patient is negative.
DETAILED DESCRIPTION OF THE INVENTION
Definitions [0040] The following definitions are provided to facilitate understanding of certain terms used throughout this specification.
[0041] In the present invention, "isolated" refers to material removed from its native environment (e.g., the natural environment if it is naturally occurring), and thus is altered "by the hand of man" from its natural state. For example, an isolated polynucleotide could be part of a vector or a composition of matter, or could be contained within a cell, and still be "isolated" because that vector, composition of matter, or particular cell is not the original environment of the polynucleotide.
[0042] In the present invention, a "membrane" C35 protein is one expressed on the cell surface through either direct or indirect association with the lipid bilayer, including, in particular, through prenylation of a carboxyl-terminal amino acid motif. Prenylation involves the covalent modification of a protein by the addition of either a farnesyl or geranylgeranyl isoprenoid. Prenylation occurs on a cysteine residue located near the carboxyl-terminus of a protein. The C35 polypeptide contains the amino acids Cys-Val-Ile-Leu at positions 112-115, with the Leu being the C terminal residue of the polypeptide. The motif Cys-X-X-Leu, where "X" represents any aliphatic amino acid, results in the addition of a 20 carbon geranylgeranyl group onto the Cys residue. Generally, following addition of this lipid the three terminal amino acid residues are cleaved off the polypeptide, and the lipid group is methylated. Prenylation promotes the membrane localization of most proteins, with sequence motifs in the polypeptide being involved in directing the prenylated protein to the plasma, nuclear, or golgi membranes.

Prenylation plays a role in protein-protein interactions, and many prenylated proteins are involved in signal transduction. Examples of prenylated proteins include Ras and the nuclear lamin B. (Zhang, F.L. and Casey, P.J., Ann. Rev.
Biochem. 65:241-269 (1996)). The C35 protein has been detected on the surface of two breast tumor cell lines by fluorescence analysis employing as a primary reagent a mouse anti-human C35 antiserum (FIGS. 4A-4C).
[0043] In the present invention, a "secreted" C35 protein refers to a protein capable of being directed to the ER, secretory vesicles, or the extracellular space as a result of a signal sequence, as well as a C35 protein released into the extracellular space without necessarily containing a signal sequence. If the secreted protein is released into the extracellular space, the C35 secreted protein can undergo extracellular processing to produce a "mature" C35 protein.
Release into the extracellular space can occur by many mechanisms, including exocytosis and proteolytic cleavage.
[0044] As used herein, a C35 "polynucleotide" refers to a molecule having a nucleic acid sequence contained in SEQ ID NO:1 . For example, the C35 polynucleotide can contain the nucleotide sequence of the full length cDNA
sequence, including the 5' and 3' untranslated sequences, the coding region, with or without the signal sequence, the secreted protein coding region, as well as fragments, epitopes, domains, and variants of the nucleic acid sequence.
Moreover, as used herein, a C35 "polypeptide" refers to a molecule having the translated amino acid sequence generated from the polynucleotide as broadly defined.
[0045] In specific embodiments, the polynucleotides of the invention are less than 300 nt, 200 nt, 100 nt, 50 nt, 15 nt, 10 nt, or 7 nt in length. In a further embodiment, polymicleotides of the invention comprise at least 15 contiguous nucleotides of C35 coding sequence, but do not comprise all or a portion of any C35 intron. In another embodiment, the nucleic acid comprising C35 coding sequence does not contain coding sequences of a genomic flanking gene (i.e., 5' or 3' to the C35 gene in the genome).

[0046] In the present invention, the full length C35 coding sequence is identified as SEQ ID NO: 1.
[0047] A C35 "polynucleotide" also refers to isolated polynucleotides which encode the C35 polyp eptides, and polynucleotides closely related thereto.
[0048] A C35 "polynucleotide" also refers to isolated polynucleotides which encode the amino acid sequence shown in SEQ ID NO: 2, or a biologically active fragment thereof.
[0049] A C35 "polynucleotide" also includes those polynucleotides capable of hybridizing, under stringent hybridization conditions, to sequences contained in SEQ ID NO: 1, the complement thereof, or the cDNA within the deposited clone.
"Stringent hybridization conditions" refers to an overnight incubation at 42 C in a solution comprising 50% formamide, 5x SSC (750 mM NaCl, 75 mM sodium citrate), 50 mM sodium phosphate (pH 7.6), 5x Denhardt's solution, 10% dextran sulfate, and 20 g/ml denatured, sheared salmon sperm DNA, followed by washing the filters in 0.1x SSC at about 65 C.
[0050] Of course, a polynucleotide which hybridizes only to polyA+
sequences (such as any 3' terminal polyA+ tract of a cDNA), or to a complementary stretch =of T (or U) residues, would not be included in the definition of "polynucleotide,"
since such a polynucleotide would hybridize to any nucleic acid molecule containing a poly (A) stretch or the complement thereof (e.g., practically any double-stranded cDNA clone).
[0051] The C35 polynucleotide can be composed of any polyribonucleotide or polydeoxyribonucleotide, which may be unmodified RNA or DNA or modified RNA or DNA. For example, C35 polynucleotides can be composed of single-and double-stranded DNA, DNA that is a mixture of single- and double-stranded regions, single- and double-stranded RNA, and RNA that is mixture of single-and double-stranded regions, hybrid molecules comprising DNA and RNA that maybe single-stranded or, more typically, double-stranded or a mixture of single-and double-stranded regions. In addition, the C35 polynucleotides can be composed of triple-stranded regions comprising RNA or DNA or both RNA and DNA. C35 polynucleotides may also contain one or more modified bases or DNA or RNA backbones modified for stability or for other reasons. "Modified"
bases include, for example, tritylated bases and unusual bases such as inosine.
A variety of modifications can be made to DNA and RNA; thus, "polynucleotide"
embraces chemically, enzymatically, or metabolically modified forms.
[0052] C35 polypeptides can be composed of amino acids joined to each other by peptide bonds or modified peptide bonds, i.e., peptide isosteres, and may contain amino acids other than the 20 gene-encoded amino acids. The C35 polypeptides may be modified by either natural processes, such as posttranslational processing, or by chemical modification techniques which are well known in the art. Such modifications are well described in basic texts and in more detailed monographs, as well as in a voluminous research literature.
Modifications can occur anywhere in the C35 polypeptide, including the peptide backbone, the amino acid side-chains and the amino or carboxyl termini. It will be appreciated that the same type of modification may be present in the same or varying degrees at several sites in a given C35 polypeptide. Also, a given C35 polypeptide may contain many types of modifications. C35 polypeptides may be branched, for example, as a result of ubiquitination, and they may be cyclic, with or without branching. Cyclic, branched, and branched cyclic C35 polypeptides may result from posttranslation natural processes or may be made by synthetic methods.
Modifications include acetylation, acylation, ADP-ribosylation, amidation, covalent attachment of Ravin, covalent attachment of a heme moiety, covalent attachment of a nucleotide or nucleotide derivative, covalent attachment of a lipid or lipid derivative, covalent attachment of phosphotidylinositol, cross-linking, cyclization, disulfide bond formation, demethylation, formation of covalent cross-links, formation of cysteine, formation of pyroglutamate, formylation, gamma-carboxylation, glycosylation, GPI anchor formation, hydroxylation, iodination, methylation, myristoylation, oxidation, pegylation, proteolytic processing, phosphorylation, prenylation, racemization, selenoylation, sulfation, transfer-RNA mediated addition of amino acids to proteins such as arginylation, and ubiquitination. (See, for instance, Proteins - Structure And Molecular Properties, 2nd Ed., T. E. Creighton, W. H.

Freeman and Company, New York (1993); Posttranslational Covalent Modification of Proteins, B. C. Johnson, Ed., Academic Press, New York, pgs.
1-12 (1983); Seifter et al., Meth Enzymol 182:626-646 (1990); Rattan et al., Ann NY Acad Sci 663:48-62 (1992).) [0053] "SEQ ID NO: 1" refers to a C35 polynucleotide sequence while "SEQ
ID
NO: 2" refers to a C35 polypeptide sequence.
[0054] A C35 polypeptide "having biological activity" refers to polypeptides exhibiting activity similar to, but not necessarily identical to, an activity of a C35 polypeptide, including mature forms, as measured in a particular biological assay, with or without dose dependency. In the case where dose dependency does exist, it need not be identical to that of the C35 polypeptide, but rather substantially similar to the dose-dependence in a given activity as compared to the C35 polypeptide (i.e., the candidate polypeptide will exhibit greater activity or not more than about 25-fold less and, preferably, not more than about tenfold less activity, and most preferably, not more than about three-fold less activity relative to the C35 polypeptide.) C35 Polynucleotides and Polypeptides [0055] A 348 base pair fragment of C35 was initially isolated by subtractive hybridization of poly-A RNA from tumor and normal mammary epithelial cell lines derived from the same patient with primary and infiltrating intraductal mammary carcinoma. Band, V. et al., Cancer Res. 50:7351-7357 (1990).
Employing primers based on this sequence and that of an overlapping EST
sequence (Accession No. W57569), a cDNA that includes the full-length C35 coding sequence was then amplified and cloned from the BT-20 breast tumor cell line (ATCC, HTB-19). This C35 cDNA contains the entire coding region identified as SEQ lD NO:l. The C35 clone includes, in addition to the 348 bp coding sequence, 167 bp of 3' untranslated region. The open reading frame begins at an N-terminal methionine located at nucleotide position 1, and ends at a stop codon at nucleotide position 348 (FIG. 1A). A representative clone containing all or most of the sequence for SEQ ID NO:1 was deposited with the American Type Culture Collection ("ATCC") on August 1,2000, and was given the ATCC Deposit Number PTA-2310. The ATCC is located at 10801 University Boulevard, Manassas, VA 20110-2209, USA. The ATCC deposit was made pursuant to the terms of the Budapest Treaty on the international recognition of the deposit of microorganisms for purposes of patent procedure.
[0056] Therefore, SEQ ID NO: 1 and the translated SEQ ID NO: 2 are sufficiently accurate and otherwise suitable for a variety of uses well known in the art and described further below. For instance, SEQ ID NO: 1 is useful for designing nucleic acid hybridization probes that will detect nucleic acid sequences contained in SEQ ID NO: 1 or the cDNA contained in the deposited clone. These probes will also hybridize to nucleic acid molecules in biological samples, thereby enabling a variety of forensic and diagnostic methods of the invention. Similarly, polypeptides identified from SEQ ID NO:2 may be used to generate antibodies which bind specifically to C35, or to stimulate T cells which are specific for C35 derived peptide epitopes in association with MHC
molecules on the cell surface.
[0057] Nevertheless, DNA sequences generated by sequencing reactions can contain sequencing errors. The errors exist as misidentified nucleotides, or as insertions or deletions of nucleotides in the generated DNA sequence. The erroneously inserted or deleted nucleotides cause frame shifts in the reading frames of the predicted amino acid sequence. In these cases, the predicted amino acid sequence diverges from the actual amino acid sequence, even though the generated DNA sequence may be greater than 99.9% identical to the actual DNA
sequence (for example, one base insertion or deletion in an open reading frame of over 1000 bases).

[0058] Accordingly, for those applications requiring precision in the nucleotide sequence or the amino acid sequence, the present invention provides not only the generated nucleotide sequence identified as SEQ ID NO:1 and the predicted translated amino acid sequence identified as SEQ ID NO:2. The nucleotide sequence of the deposited C35 clone can readily be determined by sequencing the deposited clone in accordance with known methods. The predicted C35 amino acid sequence can then be verified from such deposits. Moreover, the amino acid sequence of the protein encoded by the deposited clone can also be directly determined by peptide sequencing or by expressing the protein in a suitable host cell containing the deposited human C35 cDNA, collecting the protein, and determining its sequence.
[0059] The present invention also relates to the C35 gene corresponding to SEQ
ID NO:1, or the deposited clone. The C35 gene can be isolated in accordance with known methods using the sequence information disclosed herein. Such methods include preparing probes or primers from the disclosed sequence and identifying or amplifying the C35 gene from appropriate sources of genomic material.
[0060] Also provided in the present invention are species homologs of C35.
Species homologs may be isolated and identified by making suitable probes or primers from the sequences provided herein and screening a suitable nucleic acid source for the desired homologue.
[0061] By "C35 polypeptide(s)" is meant all forms of C35 proteins and polypeptides described herein. The C35 polypeptides can be prepared in any suitable manner. Such polyp eptides include isolated naturally occurring polypeptides, recombinantly produced polypeptides, synthetically produced polypeptides, or polypeptides produced by a combination of these methods.
Means for preparing such polypeptides are well understood in the art.
[0062] The C35 polypeptides may be in the form of the membrane protein or a secreted protein, including the mature form, or may be a part of a larger protein, such as a fusion protein (see below). It is often advantageous to include an additional amino acid sequence which contains secretory or leader sequences, pro-sequences, sequences which aid in purification, such as multiple histidine residues, or an additional sequence for stability during recombinant production.
[0063] C35 polypeptides are preferably provided in an isolated form, and preferably are substantially purified. A recombinantly produced version o f a polypeptide, including the secreted polypeptide, can be substantially purified by the one-step method described in Smith and Johnson, Gene 67:31-40 (1988).
C35 polypeptides also can be purified from natural or recombinant sources using antibodies of the invention raised against the C35 protein in methods which are well known in the art.
[0064] In one embodiment, the present invention is directed to an isolated polypeptide capable of eliciting a cytotoxic T lymphocyte and/or helper T
lymphocyte response in a human subject, the isolated polypeptide comprising, or, alternatively, consisting of, one or more C35 peptide epitopes or C35 peptide epitope analogs. In a preferred embodiment, said one or more C35 peptide epitopes are selected from the group consisting of: amino acids E4 to P12 of SEQ
ID NO:2, amino acids S9 to V17 of SEQ ID NO:2, amino acids S21 to Y29 of SEQ ID NO:2, G22 to C30 of SEQ ID NO:2, amino acids 125 to C33 of SEQ ID
NO:2, amino acids T38 to V46 of SEQ ID NO:2, amino acids G61 to 169 of SEQ
ID NO:2, amino acids T62 to N70 of SEQ ID NO:2, amino acids G63 to G71 of SEQ ID NO:2, amino acids F65 to L73 of SEQ ID NO:2, amino acids 167 to F75 of SEQ ID NO:2, amino acids K77 to Y85 of SEQ ID NO:2, amino acids Q72 to E86 of SEQ ID NO:2, amino acids G81 to L89 of SEQ ID NO:2, amino acids K104 to C112 of SEQ ID NO:2, amino acids K104 to V113 of SEQ ID NO:2, amino acids 1105 to V113 of SEQ ID NO:2, and amino acids N107 to L115 of SEQ ID NO:2. In a preferred embodiment, the isolated polypeptides comprising one or more C35 peptide epitopes (e.g., one or more octamers, nonamers, decamers, 15mers, or 20mers in Tables 1-3 or 5-6) or C35 peptide epitope analogs (e.g., an analog listed in Table 4) are not more than 114 amino acids in length, more preferably not more than 110 amino acids in length, more preferably not more than 105 amino acids in length, more preferably not more than 100 amino acids in length, more preferably not more than 95 amino acids in length, more preferably not more than 90 amino acids in length, more preferably not more than 85 amino acids in length, more preferably not more than 80 amino acids in length, more preferably not more than 75 amino acids in length, more preferably not more than 70 amino acids in length, more preferably not more than 65 amino acids in length, more preferably not more than 60 amino acids in length, more preferably not more than 55 amino acids in length, more preferably not more than 50 amino acids in length, more preferably not more than 45 amino acids in length, more preferably not more than 40 amino acids in length, more preferably not more than 35 amino acids in length, more preferably not more than 30 amino acids in length, more preferably not more than 25 amino acids in length, more preferably 20 amino acids in length, more preferably 15 amino acids in length, more preferably 14, 13, 12, 11, 10, 9 or 8 amino acids in length. Of course, although not explicitly listed here, isolated polypeptides of any length between, for example, 8 and 100 amino acids, comprising C35 peptide epitopes or C35 peptide epitope analogs are likewise contemplated by the present invention. In a preferred embodiment, the isolated polypeptide is a fragment of the C35 polypeptide shown in SEQ ID NO:2 and FIG. 1B. In another embodiment, the present invention is directed to an isolated polypeptide capable of eliciting a cytotoxic T lymphocyte and/or helper T lymphocyte response in a human subject, the isolated polypeptide comprising, or, alternatively, consisting of multiple C35 peptide epitopes. In a particularly preferred embodiment, said multi-epitopepolypeptide is selected from the group consisting of: amino acids T101 to V113 of SEQ ID NO:2, amino acids E100 to V113 of SEQ ID NO:2, amino acids G99 to V113 of SEQ ID NO:2, amino acids 193 to V113 of SEQ ID
NO:2, amino acids D88 to V113 of SEQ ID NO:2, amino acids P84 to V113 of SEQ ID NO:2, amino acids K77 to V113 of SEQ ID NO:2, amino acids Q72 to V113 of SEQ ID NO:2, amino acids F65 to V113 of SEQ ID NO:2, and amino acids L59 to V113 of SEQ ID NO:2. In another preferred embodiment, the present invention is directed to a fusion protein comprising at least one C35 peptide epitope listed in Tables 1-3 or 5-6, or a C35 peptide epitope analog listed in Table 4. In one embodiment, the at least one C35 peptide epitope or C35 peptide epitope analog is fused to a heterologous (i.e., non-C35) polypeptide.
In another preferred embodiment, said fusion protein comprises two or more C35 peptide epitopes or two or more C35 peptide epitope analogs, either as a homopolymer or a heteropolymer. In another preferred embodiment, the fusion proteins of the present invention comprise at least one C35 peptide epitope analog joined to at least one C35 peptide epitope. In a further embodiment, the epitopes/analogs are joined by an amino acid spacer or linker.
100651 The present invention is further directed to a pharmaceutical composition for use as a vaccine comprising such isolated polypeptides and fusion proteins.
[0066] The present invention is further directed to a method for stimulating a cytotoxic T lymphocyte and/or a helper T lymphocyte response in a human patient comprising administering to said patient an immunogenically effective amount of the pharmaceutical composition of the invention.
Polynucleotide and Polypeptide Variants [0067] "Variant" refers to a polynucleotide or polypeptide differing from the C35 polynucleotide or polypeptide, but retaining essential properties thereof.
Generally, variants are overall closely similar, and, in many regions, identical to the C35 polynucleotide or polypeptide.
[0068] By a polynucleotide having a nucleotide sequence at least, for example, 95% "identical" to a reference nucleotide sequence of the present invention, it is intended that the nucleotide sequence of the polynucleotide is identical to the reference sequence except that the polynucleotide sequence may include up to five point mutations per each 100 nucleotides of the reference nucleotide sequence encoding the C35 polypeptide. In other words, to obtain a polynucleotide having a nucleotide sequence at least 95% identical to a reference nucleotide sequence, up to 5% of the nucleotides in the reference sequence may be deleted or substituted with another nucleotide, or a number of nucleotides up to 5% of the total nucleotides in the reference sequence may be inserted into the reference sequence. The query sequence may be an entire sequence shown of SEQ ID NO:1, the ORF (open reading frame), or any fragment specified as described herein.
[0069] As a practical matter, whether any particular nucleic acid molecule or polypeptide is at least 90%, 95%, 96%, 97%, 98% or 99% identical to a nucleotide sequence or polypeptide sequence of the presence invention can be determined conventionally using known computer programs. A preferred method for determining the best overall match between a query sequence (a sequence of the present invention) and a subject sequence, also referred to as a global sequence alignment, can be determined using the FASTDB computer program based on the algorithm of Brutlag et al., Comp. App. Biosci. 6:237-245 (1990).

In a sequence alignment the query and subject sequences are both DNA
sequences. An RNA sequence can be compared by converting U's to T's. The result of said global sequence alignment is in percent identity. Preferred parameters used in a FASTDB alignment of DNA sequences to calculate percent identity are: Matrix=Unitary, k-tuple=4, Mismatch Penalty=1, Joining Penalty=30, Randomization Group Length=0, Cutoff Score=1, Gap Penalty=5, Gap Size Penalty 0.05, Window Size=500 or the length of the subject nucleotide sequence, whichever is shorter.
[0070] If the subject sequence is shorter than the query sequence because of 5' or 3' deletions, not because of internal deletions, a manual correction must be made to the results. This is because the FASTDB program does not account for 5' and 3' truncations of the subject sequence when calculating percent identity. For subject sequences truncated at the 5' or 3' ends, relative to the query sequence, the percent identity is corrected by calculating the number of bases of the query sequence that are 5' and 3' of the subject sequence, which are not matched/aligned, as a percent of the total bases of the query sequence.
Whether a nucleotide is matched/aligned is determined by results of the FASTDB
sequence alignment. This percentage is then subtracted from the percent identity, calculated by the above FASTDB program using the specified parameters, to arrive at a final percent identity score. This corrected score is what is used for the purposes of the present invention. Only bases outside the 5' and 3' bases of the subject sequence, as displayed by the FASTDB alignment, which are not matched/aligned with the query sequence, are calculated for the purposes of manually adjusting the percent identity score.
[0071] For example, a 90 base subject sequence is aligned to a 100 base query sequence to determine percent identity. The deletions occur at the 5' end of the subject sequence and therefore, the FASTDB alignment does not show a matched/alignment of the first 10 bases at 5' end. The 10 unpaired bases represent 10% of the sequence (number of bases at the 5' and 3' ends not matched/total number of bases in the query sequence) so 10% is subtracted from the percent identity score calculated by the FASTDB program. If the remaining 90 bases were perfectly matched the final percent identity would be 90%. In another example, a 90 base subject sequence is compared with a 100 base query sequence. This time the deletions are internal deletions so that there are no bases on the 5' or 3' of the subject sequence which are not matched/aligned with the query. In this case the percent identity calculated by FASTDB is not manually corrected. Once again, only bases 5' and 3' of the subject sequence which are not matched/aligned with the query sequence are manually corrected for. No other manual corrections are to made for the purposes of the present invention.
[0072] By a polypeptide having an amino acid sequence at least, for example, 95% "identical" to a query amino acid sequence of the present invention, it is intended that the amino acid sequence of the subject polypeptide is identical to the query sequence except that the subject polypeptide sequence may include up to five amino acid alterations per each 100 amino acids of the query amino acid sequence. In other words, to obtain a polypeptide having an amino acid sequence at least 95% identical to a query amino acid sequence, up to 5% of the amino acid residues in the subject sequence may be inserted, deleted, or substituted with another amino acid. These alterations of the reference sequence may occur at the amino or carboxy terminal positions of the reference amino acid sequence or anywhere between those terminal positions, interspersed either individually among residues in the reference sequence or in one or more contiguous groups within the reference sequence.
[0073] As a practical matter, whether any particular polypeptide is at least 90%, 95%, 96%, 97%, 98% or 99% identical to, for instance, the amino acid sequence shown in SEQ ID NO:2 or to the amino acid sequence encoded by deposited DNA clone, can be determined conventionally using known computer programs.
A preferred method for determining the best overall match between a query sequence (a sequence of the present invention) and a subject sequence, also referred to as a global sequence alignment, can be determined using the FASTDB

computer program based on the algorithm of Brutlag et al., Comp. App. Biosei.
6:237-245 (1990). In a sequence alignment the query and subject sequences are either both nucleotide sequences or both amino acid sequences. The result of said global sequence alignment is in percent identity. Preferred parameters used in a FASTDB amino acid alignment are: Matrix=PAM 0, k-tuple=2, Mismatch Penalty=1, Joining Penalty=20, Randomization Group Length=0, Cutoff Score=1, Window Size=sequence length, Gap Penalty=5, Gap Size Penalty=0.05, Window Size=500 or the length of the subject amino acid sequence, whichever is shorter.
[0074] If the subject sequence is shorter than the query sequence due to ¨
or C-terminal deletions, not because of internal deletions, a manual correction must be made to the results. This is because the FASTDB program does not account for ¨ and C-terminal truncations of the subject sequence when calculating global percent identity. For subject sequences truncated at the ¨ and C-termini, relative to the query sequence, the percent identity is corrected by calculating the number of residues of the query sequence that are ¨ and C-terminal of the subject sequence, which are not matched/aligned with a corresponding subject residue, as a percent of the total bases of the query sequence. Whether a residue is matched/aligned is determined by results of the FASTDB sequence alignment.
This percentage is then subtracted from the percent identity, calculated by the above FASTDB program using the specified parameters, to arrive at a final percent identity score. This fmal percent identity score is what is used for the purposes of the present invention. Only residues to the ¨ and C-termini of the subject sequence, which are not matched/aligned with the query sequence, are considered for the purposes ofmanually adjusting the percent identity score.
That is, only query residue positions outside the farthest ¨ and C-terminal residues of the subject sequence.
[0075] For example, a 90 amino acid residue subject sequence is aligned with a 100 residue query sequence to determine percent identity. The deletion occurs at the N-terminus of the subject sequence and therefore, the FASTDB alignment does not show a matching/alignment of the first 10 residues at the N-terminus.

The 10 unpaired residues represent 10% of the sequence (number of residues at the ¨ and C- termini not matched/total number of residues in the query sequence) so 10% is subtracted from the percent identity score calculated by the FASTDB
program. If the remaining 90 residues were perfectly matched the final percent identity would be 90%. In another example, a 90 residue subject sequence is compared with a 100 residue query sequence. This time the deletions are internal deletions so there are no residues at the ¨ or C-termini of the subject sequence which are not matched/aligned with the query. In this case the percent identity calculated by FASTDB is not manually corrected. Once again, only residue positions outside the¨ and C-terminal ends of the subject sequence, as displayed in the FASTDB alignment, which are not matched/aligned with the query sequence are manually corrected for. No other manual corrections are to be made for the purposes of the present invention.
[0076] The C35 variants may contain alterations in the coding regions, non-coding regions, or both. Especially preferred are polynucleotide variants containing alterations which produce silent substitutions, additions, or deletions, but do not alter the properties or activities of the encoded polypeptide.

Nucleotide variants produced by silent substitutions due to the degeneracy of the genetic code are preferred. Moreover, variants in which 5-10, 1-5, or 1-2 amino acids are substituted, deleted, or added in any combination are also preferred.
C35 polynucleotide variants can be produced for a variety of reasons, e.g., to optimize codon expression for a particular host (change codons in the human mRNA to those preferred by a bacterial host such as E. co/i).
[0077] Naturally occurring C35 variants are called "allelic variants,"
and refer to one of several alternate forms of a gene occupying a given locus on a chromosome of an organism. (Genes II, Lewin, B., ed., John Wiley & Sons, New York (1985).) Also, allelic variants can occur as "tandem alleles" which are highly homologous sequences that occur at different loci on chromosomes of an organism. These allelic variants can vary at either the polynucleotide and/or polypeptide level. Alternatively, non-naturally occurring variants may be produced by mutagenesis techniques or by direct synthesis.
[0078] Using known methods of protein engineering and recombinant DNA
technology, variants may be generated to improve or alter the characteristics of the C35 polypeptides. For instance, one or more amino acids can be deleted from the N-terminus or C-terminus of the secreted protein without substantial loss of biological function. The authors of Ron et al., J. Biol. Chem. 268: 2984-2988 (1993), reported variant KGF proteins having heparin binding activity even after deleting 3, 8, or 27 amino-terminal amino acid residues. Similarly, Interferon gamma exhibited up to ten times higher activity after deleting 8-10 amino acid residues from the carboxy terminus of this protein (Dobeli et al., J.
Biotechnology 7:199-216 (1988)).
[0079] Moreover, ample evidence demonstrates that variants often retain a biological activity similar to that of the naturally occurring protein. For example, Gayle and coworkers (J. Biol. Chem 268:22105-22111 (1993)) conducted extensive mutational analysis of human cytokine IL-la. They used random mutagenesis to generate over 3,500 individual IL-la mutants that averaged 2.5 amino acid changes per variant over the entire length of the molecule.
Multiple mutations were examined at every possible amino acid position. The investigators found that "[m]ost of the molecule could be altered with little effect on either [binding or biological activity]." (See, Abstract.) In fact, only 23 unique amino acid sequences, out of more than 3,500 nucleotide sequences examined, produced a protein that significantly differed in activity from wild-type.
[0080] Furthermore, even if deleting one or more amino acids from the N-terminus or C-terminus of a polypeptide results in modification or loss of one or more biological functions, other biological activities may still be retained. For example, the ability of a deletion variant to induce and/or to bind antibodies which recognize the secreted form will likely be retained when less than the majority of the residues of the secreted form are removed from the N-terminus or C-terminus. Whether a particular polypeptide lacking ¨ or C-terminal residues of a protein retains such immunogenic activities can readily be determined by routine methods described herein and otherwise known in the art.
[0081] Thus, the invention further includes C35 polyp eptide variants which show substantial biological activity. Such variants include deletions, insertions, = inversions, repeats, and substitutions selected according to general rules known in the art so as to have little effect on activity. For example, guidance concerning how to make phenotypically silent amino acid substitutions is provided in Bowie, J. U. et al., Science 247:1306-1310(1990), wherein the authors indicate that there are two main strategies for studying the tolerance of an amino acid sequence to change.
[0082] The first strategy exploits the tolerance of amino acid substitutions by natural selection during the process of evolution. By comparing amino acid sequences in different species, conserved amino acids can be identified. These conserved amino acids are likely important for protein function. In contrast, the amino acid positions where substitutions have been tolerated by natural selection indicates that these positions are not critical for protein function. Thus, positions tolerating amino acid substitution could be modified while still maintaining biological activity of the protein.
[0083] The second strategy uses genetic engineering to introduce amino acid changes at specific positions of a cloned gene to identify regions critical for protein function. For example, site directed mutagenesis or alanine-scanning mutagenesis (introduction of single alanine mutations at every residue in the molecule) can be used. (Cunningham and Wells, Science 244:1081-1085(1989).) The resulting mutant molecules can then be tested for biological activity.
10084] As the authors state, these two strategies have revealed that proteins are surprisingly tolerant of amino acid substitutions. The authors further indicate which amino acid changes are likely to be permissive at certain amino acid positions in the protein. For example, most buried (within the tertiary structure of the protein) amino acid residues require nonpolar side chains, whereas few features of surface side chains are generally conserved. Moreover, tolerated conservative amino acid substitutions involve replacement of the aliphatic or hydrophobic amino acids Ala, Val, Leu and Ile; replacement of the hydroxyl residues Ser and Tin; replacement of the acidic residues Asp and Glu;
replacement of the amide residues Asn and Gin, replacement of the basic residues Lys, Arg, and His; replacement of the aromatic residues Phe, Tyr, and Trp, and replacement of the small-sized amino acids Ala, Ser, Tin, Met, and Gly.
[0085] Besides conservative amino acid substitution, variants of C35 include (i) substitutions with one or more of the non-conserved amino acid residues, where the substituted amino acid residues may or may not be one encoded by the genetic code, or (ii) substitution with one or more of amino acid residues having a substituent group, or (iii) fusion of the mature polypeptide with another compound, such as a compound to increase the stability and/or solubility of the polypeptide (for example, polyethylene glycol), or (iv) fusion of the polypeptide with additional amino acids, such as an IgG Fc fusion region peptide, or leader or secretory sequence, or a sequence facilitating purification. Such variant polypeptides are deemed to be within the scope of those skilled in the art from the teachings herein.
[0086] For example, C35 polypeptide variants containing amino acid substitutions of charged amino acids with other charged or neutral amino acids may produce proteins with improved characteristics, such as less aggregation.
Aggregation of pharmaceutical formulations both reduces activity and increases clearance due to the aggregate's immunogenic activity. (Pinckard et al., Clin.

Exp. Immunol. 2:331-340 (1967); Robbins et al., Diabetes 36: 838-845 (1987);
Cleland et al., Grit. Rev. Therapeutic Drug Carrier Systems 10:307-377(1993).) Polynucleotide and Polypeptide Fragments [0087] In the present invention, a "polynucleotide fragment" refers to a short polynucleotide having a nucleic acid sequence contained in the deposited clone or shown in SEQ ID NO: 1. The short nucleotide fragments are preferably at least about 15 nt, and more preferably at least about 20 nt, still more preferably at least about 30 nt, and even more preferably, at least about 40 nt in length. A
fragment "at least 20 nt in length," for example, is intended to include 20 or more contiguous bases from the cDNA sequence contained in the deposited clone or the nucleotide sequence shown in SEQ ID NO: 1. These nucleotide fragments are useful as diagnostic probes and primers as discussed herein. Of course, larger fragments (e.g., at least 50, 100, 150, 200, 250, 300 nucleotides) are preferred.
[0088] Moreover, representative examples of C35 polynucleotide fragments include, for example, fragments having a sequence from about nucleotide number 1-50, 51-100,101-150, 151-200, 201-250, 251-300, or 301 to the end of SEQ ID
NO:1 or the cDNA contained in the deposited clone. In this context "about"
includes the particularly recited ranges, larger or smaller by several (5, 4, 3, 2, or 1) nucleotides, at either terminus or at both termini. Preferably, these fragments encode a polypeptide which has biological activity. More preferably, these polynucleotides can be used as probes or primers as discussed herein.

[0089] In the present invention, a "polypeptide fragment" refers to a short amino acid sequence contained in SEQ ID NO:2 and FIG. 1B or encoded by the cDNA
contained in the deposited clone. Protein fragments may be "free-standing," or comprised within a larger polypeptide of which the fragment forms a part or region, most preferably as a single continuous region. Representative examples of polypeptide fragments of the invention, include, for example, fragments from about amino acid number 1-20, 21-40, 41-60, 61-80, 81-100, or 101 to the end of the coding region. Moreover, polypeptide fragments can comprise about 7, 8, 9, 15, 20, 30, 40, 50, 60, 70, 80, 90, or 100 amino acids in length. In this context "about" includes the particularly recited ranges or lengths, larger or smaller by several (5, 4, 3, 2, or 1) amino acids, at either extreme or at both extremes.
[0090] Preferred polypeptide fragments include the secreted C35 protein as well as the mature form. Further preferred polypeptide fragments include the secreted C35 protein or the mature form having a continuous series of deleted residues from the amino or the carboxy terminus, or both. Further preferred polypeptide fragments include fragments of the C35 polypeptide comprising one or more C35 peptide epitopes.
[0091] As mentioned above, even if deletion of one or more amino acids from the N-terminus of a protein results in modification or loss of one or more biological functions of the protein, other biological activities may still be retained.
Thus, the ability of shortened C35 muteins to induce and/or bind to antibodies which recognize the complete or mature forms of the polypeptides generally will be retained when less than the majority of the residues of the complete or mature polypeptide are removed from the N-terminus. Whether a particular polypeptide lacking N-terminal residues of a complete polypeptide retains such immunologic activities can readily be determined by routine methods described herein and otherwise known in the art. It is not unlikely that a C35 mutein with a large number of deleted N-terminal amino acid residues may retain some biological or immunogenic activities. In fact, in the case of C35 peptide epitopes, peptides composed of as few as 9, 8, or even 7 C35 amino acid residues often evoke an immune response.
100921 Accordingly, the present invention further provides polypeptides having one or more residues deleted from the amino terminus of the C35 amino acid sequence shown in SEQ ID NO:2, up to the Threonine residue at position number 105 and polynucleotides encoding such polypeptides.
[0093] Also as mentioned above, even if deletion of one or more amino acids from the C-terminus of a protein results in modification or loss of one or more biological functions of the protein, other biological activities may still be retained. Thus, the ability of the shortened C35 mutein to induce cytotoxic T
lymphocytes (CTLs) and/or helper T lymphocytes (HTLs) and/or to bind to antibodies which recognize the complete or mature forms of the polypeptide generally will be retained when less than the majority of the residues of the complete or mature polypeptide are removed from the C-terminus. Whether a particular polypeptide lacking C-terminal residues of a complete polypeptide retains such immunologic activities can readily be determined by routine methods described herein and otherwise known in the art. It is not unlikely that a C35 mutein with a large number of deleted C-terminal amino acid residues may retain some biological or immunogenic activities.
[0094] Accordingly, the present invention further provides polypeptides having one or more residues deleted from the carboxy terminus of the amino acid sequence of the C35 polypeptide shown in SEQ ID NO:2, up to the valine residue at position number 10, and polynucleotides encoding such polypeptides [0095] Moreover, the invention also provides polypeptides having one or more amino acids deleted from both the amino and the carboxyl termini. hi preferred embodiments, the invention is directed to peptides having residues: E4 to P12, to V17; V10 to V17; E16 to V23; E16 to R24; E16 to 125; S21 toY29; S21 to F35; G22 to C30; 125 to C33; C30 to T38; E31 to Y39; E36 to A43; A37 to A45;
A37 to V46; T38 to V46; Y39 to V46; S44 to 153; A45 to 153; G52 to L59; E54 to T62; S57 to F75; R58 to 167; L59 to V113; G61 to 169; T62 to N70, G63 to G71, G63 to F83; F65 to L73; F65 to V113; E66 to L73; E66 to V74; 167 to F75;
K77 to Y85; K77 to V113; Q72 to E86; Q72 to V113; G81 to L89; F83 to E103;
P84 to V113; D88 to A96; D88 to V113; L89 to A96; A92 to T101;193 to V113;
R95 to L102; A96 to K104; G99 to V113, E100 to V113, T101 to V113; K104 to C112; K104 to V113; 1105 to V113; 1105 to 1114; or N107 to,L115 of SEQ
ID NO:2 and polynucleotides encoding such polypeptides.
[0096] Many polynucleotide sequences, such as EST sequences, are publicly available and accessible through sequence databases.
,[0097] The human EST sequences referred to below were identified in a BLAST
search of the EST database. These sequences are believed to be partial sequences of the cDNA inserts identified in the recited GenBank accession numbers. No homologous sequences were identified in a search of the annotated GenBank database. The Expect value (E) is a parameter that describes the number of hits one can "expect" to see just by chance when searching a database of a particular size. It decreases exponentially with the Score (S) that is assigned to a match between two sequences. Essentially, the E value describes the random background noise that exists for matches between sequences. In BLAST 2.0, the Expect value is also used instead of the P value (probability) to report the significance of matches. For example, an E value of 1 assigned to a hit can be interpreted as meaning that in a database of the current size one might expect to see 1 match with a similar score simply by chance.
[0098] For example, the following sequences are related to SEQ ID NO:1, GenBank AccessionNos.: AA971857 (SEQ ID NO:3); W57569 (SEQ ID NO:4);
AI288765 (SEQ ID NO:5 ); W65390 (SEQ ID NO:6 ); W37432 (SEQ ID NO: 7);
N42748 (SEQ ID NO:8 ); AA971638 (SEQ ID NO:9 ); R22331 (SEQ ID NO:10);
AA308370 (SEQ ID NO:11 ); AA285089 (SEQ ID NO:12 ); R68901 (SEQ ID
NO:13 ); AA037285 (SEQ ID NO:14 ); H94832 (SEQ ID NO:15); H96058 (SEQ
ID NO:16); H56522 (SEQ ID NO:17); AA935328 (SEQ ID NO: 18); AW327450 (SEQ ID NO:19); AW406075 (SEQ ID NO:20); AW406223 (SEQ ID NO:21);
M909652 (SEQ ID NO:22); AA026773 (SEQ ID NO: 23); H96055 (SEQ ID

NO:24); 1112836 (SEQ ID NO:25); R22401 (SEQ ID NO:26); N34596 (SEQ ID
NO:27); W32121 (SEQ ID NO:28); T84927 (SEQ ID NO:29); R63575 (SEQ ID
NO:30); R23139 (SEQ ID NO:31); AA337071 (SEQ ID NO:32); AA813244 (SEQ ID NO:33); AA313422 (SEQ ID NO:34); N31910 (SEQ ID NO:35);
N42693 (SEQ ID NO:36); N32532 (SEQ ID NO:37); AA375119 (SEQ ID
NO:38); R32153 (SEQ ID NO:39); R23369 (SEQ ID NO:40); AA393628 (SEQ
ID NO:41); 1112779 (SEQ ID NO:42); AI083674 (SEQ ID NO:43); AA284919 (SEQ ID NO:44); AA375286 (SEQ ID NO:45); AA830592 (SEQ ID NO:46);
1195363 (SEQ ID NO:47); T92052 (SEQ ID NO:48); A1336555 (SEQ ID
NO:49); A1285284 (SEQ ID NO:50); AA568537 (SEQ ID NO:51); AI041967 (SEQ ID NO:52); W44577 (SEQ ID NO:53); R22332 (SEQ ID NO:54); N27088 (SEQ ID NO:55); 1196418 (SEQ ID NO:56); AI025384 (SEQ ID NO:57);
AA707623 (SEQ ID NO:58); A1051009 (SEQ ID NO:59); AA026774 (SEQ ID
NO:60); W51792 (SEQ ID NO:61); A1362693 (SEQ ID NO:62); AA911823 (SEQ ID NO:63); 1196422 (SEQ ID NO:64); AI800991 (SEQ ID NO:65);
AI525314 (SEQ ID NO:66); A1934846 (SEQ ID NO:67); AI937133 (SEQ ID
NO:68); AW006797 (SEQ ID NO:69); AI914716 (SEQ ID NO:70); AI672936 (SEQ ID NO:71); W61294 (SEQ ID NO:72); AI199227 (SEQ ID NO:73);
AI499727 (SEQ ID NO:74); R32154 (SEQ ID NO:75); A1439771 (SEQ ID
NO:76); AA872671 (SEQ ID NO:77); AA502178 (SEQ ID NO:78); N26715 (SEQ ID NO:79); AA704668 (SEQ ID NO:80); R68799 (SEQ ID NO:81);
1156704 (SEQ ID NO:82); A13 60416 (SEQ ID NO:83).
[0099] Thus, in one embodiment the present invention is directed to polynucleotides comprising the polynucleotide fragments and full-length polynucleotide (e.g. the coding region) described herein exclusive of one or more of the above-recited ESTs. Also, the nucleotide sequences in SEQ ID NO: 152, SEQ ID NO: 154, and SEQ ID NO: 156 are excluded from the present invention.
[0100] Also preferred are C35 polypeptide and polynucleotide fragments characterized by structural or functional domains. Preferred embodiments of the invention include fragments that comprise MHC binding epitopes and prenylation sites.
[0101] Other preferred fragments are biologically active C35 fragments.
Biologically active fragments are those exhibiting activity similar, but not necessarily identical, to an activity of the C35 polypeptide. The biological activity of the fragments may include an improved desired activity, or a decreased undesirable activity.
Epitopes & Antibodies [0102] Cellular peptides derived by degradation of endogenously synthesized proteins are translocated into a pre-Golgi compartment where they bind to Class I or Class II MHC molecules for transport to the cell surface. These class I
MHC:peptide complexes are the target antigens for specific CD8+ cytotoxic T
cells. Since all endogenous proteins "turn over," peptides derived from any cytoplasmic or nuclear protein may bind to an MHC molecule and be transported for presentation at the cell surface. This allows T cells to survey a much larger representation of cellular proteins than antibodies which are restricted to recognize conformational determinants of only those proteins that are either secreted or integrated at the cell membrane. The T cell receptor antigen binding site interacts with determinants of both the peptide and the surrounding MEC.
T cell specificity must, therefore, be defined in terms df an MHC:peptide complex. The specificity of peptide binding to MHC molecules is very broad and of relatively low affinity in comparison to the antigen binding site of specific antibodies. Class I-bound peptides are generally 8-10 residues in length that accommodate amino acid side chains of restricted diversity at certain key positions that match pockets in the MHC peptide binding site. These key features of peptides that bind to a particular MHC molecule constitute a peptide binding motif.

[0103] The term "derived" when used to discuss a peptide epitope is a synonym for "prepared." A derived epitope can be isolated from a natural source, or it can be synthesized in accordance with standard protocols in the art. Synthetic epitopes can comprise artificial amino acids "amino acid mimetics," such as D
isomers of natural occurring L amino acids or non-natural amino acids such as cyclohexylalanine. A derived/prepared epitope can be an analog of a native epitope.
[0104] An "epitope" is the collective features of a molecule, such as primary, secondary and tertiary peptide structure, and charge, that together form a site recognized by an immunoglobulin, T cell receptor or HLA molecule.
Alternatively, an epitope can be defined as a set of amino acid residues which is involved in recognition by a particular immuno globulin, or in the context of T
cells, those residues necessary for recognition by T cell receptor proteins and/or Major Histocompatibility Complex (MHC) receptors. Epitopes are present in nature, and can be isolated, purified or otherwise prepared/derived by humans.

For example, epitopes can be prepared by isolation from a natural source, or they can be synthesized in accordance with standard protocols in the art. Synthetic epitopes can comprise artificial amino acids "amino acid mimetics," such as D
isomers of natural occurring L amino acids or non-natural amino acids such as cyclohexylalanine. Throughout this disclosure, the terms epitope and peptide are often used interchangeably. Also, the term epitope as used herein is generally understood to encompass analogs of said epitopes.
[0105] It is to be appreciated that protein or polypeptide molecules that comprise one or more C35 peptide epitopes of the invention as well as additional amino acid(s) are still within the bounds of the invention. In certain embodiments, there is a limitation on the length of a polypeptide of the invention of, for example, not more than 114 amino acids, not more than 110 amino acids, not more than 100 amino acids, not more than 95 amino acids, not more than 90 amino acids, not more than 85 amino acids, not more than 80 amino acids, not more than 75 amino acids, not more than 70 amino acids, not more than 65 amino acids, not more than 60 amino acids, not more than 55 amino acids, not more than 50 amino acids, not more than 45 amino acids, not more than 40 amino acids, not more than 35 amino acids, not more than 30 amino acids, not more than 25 amino acids, 20 amino acids, 15 amino acids, or 14, 13, 12, 11, 10, 9 or 8 amino acids. In some instances, the embodiment that is length-limited occurs when the protein/polypeptide comprising an epitope of the invention comprises a region (i.e., a contiguous series of amino acids) having 100% identity with a native sequence. In order to avoid the definition of epitope from reading, e.g., on whole natural molecules, there is a limitation on the length of any region that has 100%
identity with a native polypeptide sequence. Thus, for a polypeptide comprising an epitope of the invention and a region with 100% identity with the native polypeptide sequence, the region with 100% identity to the native sequence generally has a length of: less than or equal to 114 amino acids, more often less than or equal to 100 amino acids, often less than or equal to 85 amino acids, often less than or equal to 75 amino acids, often less than or equal to 65 amino acids, and often less than or equal to 50 amino acids. In certain embodiments, the polypeptide of the invention comprises a peptide having a region with less than 50 amino acids that has 100% identity to a native peptide sequence, in any increment of amino acids (i.e., 49, 48, 47, 46, 45, 44, 43, 42, 41, 40, 39, 38, 37, 36, 35, 34, 33, 32, 31, 30, 29, 28, 27, 26, 25, 24, 23, 22, 21, 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10, 9, 8, 7, 6, 5) down to 5 amino acids. Preferably, such C35 polypeptide comprises one or more C35 peptide epitopes.
[0106] Accordingly, polypeptide or protein sequences longer than 100 amino acids are within the scope of the invention, so long as they do not comprise any contiguous sequence of more than 114 amino acids that have 100% identity with a native polypeptide sequence. For any polypeptide that has five contiguous residues or less that correspond to a native sequence, there is no limitation on the maximal length of that polyp eptide in order to fall within the scope of the invention. In one embodiment, the polypeptide of the invention comprising one or more C35 peptide epitopes is less than 60 residues long in any increment down to eight amino acid residues.
[0107] An "immunogenic peptide" or "peptide epitope" is a peptide that will bind an HA molecule and induce a cytotoxic T lymphocyte (CTL) response and/or a helper T lymphocyte (HTL) response. Thus, immunogenic peptides of the invention are capable of binding to an appropriate HLA molecule and thereafter inducing a cytotoxic T lymphocyte (CTL) response, or a helper T lymphocyte (HTL) response, to the peptide.
[0108] The term "motif' refers to a pattern of residues in an amino acid sequence of defined length, usually a peptide of from about 8 to about 13 amino acids for a class I HLA motif and from about 16 to about 25 amino acids for a class II
HLA
motif, which is recognized by a particular HLA molecule. Motifs are typically different for each HLA protein encoded by a given human HLA allele. These motifs often differ in their pattern of the primary and secondary anchor residues.
[0109] A
"protective immune response" or "therapeutic immune response" refers to a cytotoxic T lymphocyte (CTL) and/or an helper T lymphocyte (HTL) response to an antigen derived from an pathogenic antigen (e.g., an antigen from an infectious agent or a tumor antigen), which in some way prevents or at least partially arrests disease symptoms, side effects or progression. The immune response may also include an antibody response which has been facilitated by the stimulation of helper T cells.
[0110] The term "residue" refers to an amino acid or amino acid mimetic incorporated into a peptide or protein by an amide bond or amide bond mimetic.
[0111] "Synthetic peptide" refers to a peptide that is not naturally occurring, but is man-made using such methods as chemical synthesis or recombinant DNA
technology.
[0112] As used herein, a "vaccine" is a composition that contains one or more peptide epitopes of the invention, see, e.g., Tables 1-3 and 5-6, exclusive of peptide E-100 to R-109, and a pharmaceutically acceptable carrier. There are numerous embodiments of vaccines in accordance with the invention, such as by a cocktail of one or more peptides; a polyepitopic peptide comprising one or more peptides of the invention; or nucleic acids that encode such peptides or polypeptides, e.g., a minigene that encodes a polyepitopic peptide. The "one or more peptides" or "one or more epitopes" can include, for example, at least 2, 3, 4, 5, 6, 7, 8,9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34,35, 36, 37, 38, 39, 40, 45, 50, 55, 60, 65, 70,75, 80, 85, 90, 95, or 100 or more peptides or epitopes of the invention. The peptides or polypeptides can optionally be modified, such as by lipidation, addition of targeting or other sequences. HLA class I-binding peptides of the invention can be linked to HLA class II-binding peptides, to facilitate activation of both cytotoxic T lymphocytes and helper T lymphocytes. Vaccines can comprise peptide pulsed antigen presenting cells, e.g., dendritic cells.
[0113] In a preferred embodiment, the isolated polypeptides of the present invention comprise or, alternatively, consist of one or more of the following peptide epitopes: amino acids E4 to P12 of SEQ ID NO:2, amino acids S9 to V17 of SEQ ID NO:2, amino acids S21 to Y29 of SEQ ID NO:2, G22 to C30 of SEQ
ID NO: 2, amino acids 125 to C33 of SEQ ID NO:2, amino acids T38 to V46 of SEQ ID NO:2, amino acids G61 to 169 of SEQ ID NO:2, amino acids T62 to N70 of SEQ ID NO:2, amino acids G63 to G71 of SEQ ID NO:2, amino acids F65 to L73 of SEQ ID NO:2, amino acids 167 to F75 of SEQ ID NO:2, amino acids K77 to Y85 of SEQ ID NO:2, amino acids Q72 to E86 of SEQ ID NO:2, amino acids G81 to L89 of SEQ ID NO:2, amino acids K104 to C112 of SEQ ID NO:2, amino acids K104 to V113 of SEQ ID NO:2, amino acids 1105 to V113 of SEQ ID
NO:2, or amino acids N107 to L115 of SEQ ID NO:2. In another embodiment, said polypeptides comprising or, alternatively, consisting of one or more C35 peptide epitopes are selected from the group consisting of: T101 to V113 of SEQ
ID NO:2, G99 to V113 of SEQ ID NO:2, E100 to V113 of SEQ ID NO:2, 193 to V113 of SEQ ID NO:2, D88 to V113 of SEQ ID NO:2, P84 to V113 of SEQ ID
NO:2, K77 to V113 of SEQ ID NO:2, Q72 to V113 of SEQ ID NO:2, F65 to V113 of SEQ ID NO:2, and L59 to V113 of SEQ ID NO:2. It is contemplated that fragments of C35 peptide epitopes and polypeptides comprising fragments of C35 peptide epitopes of the invention will, in some instances, also be useful for stimulating a cytotoxic T lymphocyte response. Thus, the present invention includes fragments of the C35 peptide epitopes in which 1, 2, 3, 4, 5 or more amino acids of the peptide sequence provided have been deleted from either the amino terminus or the carboxy terminus of the peptide. In addition, it is contemplated that larger fragments of the C35 polypeptide that contain one or more of the peptide epitopes of the invention may also be used to stimulate a CTL
response in a patient. It is further contemplated that polypeptides that comprise one or more peptide epitopes of the present invention in addition to heterologous, i.e., non-C35, flanking sequences may also be used to stimulate a CTL
response.
[0114] In addition to the specific C35 peptide epitopes specifically listed above, many other peptide epitopes are contemplated by the present invention. Thus, the isolated polypeptides of the present invention comprising or, alternatively, consisting of, one or more C35 peptide epitopes include the following 8mers (residues correspond to SEQM NO:2 and FIG. 1B): M1 to T8; S2 to S9; G3 to V10; E4 to All; P5 to P12; G6 to P13; Q7 to P14; T8 to E15; S9 to E16; V10 to V17; All to E18; P12 to P19; P13 to G20; P14 to S21; E15 to G22; E16 to V23;
V17 to R24; El8 to 125; P19 to V26; G20 to V27; S21 to E28; G22 to Y29; V23 to C30; R24 to E31; 125 to P32; V26 to C33; V27 to G34; E28 to F35; Y29 to E36; C30 to A37; E31 to T38; P32 to Y39; C33 to MO; G34 to E41; F35 to L42;
E36 to A43; A37 to S44; T38 to A45; Y39 to V46; L40 to K47; E41 to E48; L42 to Q49; A43 to Y50; S44 to P51; A45 to G52; V46 to 153; K47 to E54; E48 to 155; Q49 to E56; Y50 to S57; P51 to R58; G52 to L59; 153 to G60; E54 to G61;
155 to T62; E56 to G63; S57 to A64; R58 to F65; L59 to E66; G60 to 167; G61 to E68; T62 to 169; G63 to N70; A64 to G71; F65 to Q72; E66 to L73; 167 to V74; E68 to F75; 169 to S76; N70 to K77; G71 to L78; Q72 to E79; L73 to N80;
V74 to G81; F75 to G82; S76 to F83; K77 to P84; L78 to Y85; E79 to E86; N80 to K87; G81 to D88; G82 to L89; F83 to 190; P84 to E91; y85 to A92; E86- to 193; K87 to R94; D88 to R95; L89 to A96; 190 to S97; E91 to N98; A92 to G99;

193 to E100; R94 to T101; R95 to L102; A96 to E103; S97 to K104; N98 to 1105;
G99 to T106; E100 to N107; T101 to S108; L102 to R109; E103 to P110; K104 to P111; 1105 to C112; T106 to V113; N107 to I114; and S108 to L115.
[0115] In a further embodiment, the isolated polypeptides ofthe present invention comprising or, alternatively, consisting of, one or more C35 peptide epitopes include the following 9mers (residues correspond to SEQ ID NO:2 and FIG. 1B):
M1 to S9; S2 to V10; G3 to All; E4 to P12; P5 to P13; G6 to P14; Q7 to E15;
T8 to E16; S9 to V17; V10 to E18; All to P19; P12 to G20; P13 to S21; P14 to G22; E15 to V23; El6 to R24; V17 to 125; E18 to V26; P19 to V27; G20 to E28;
S21 to Y29; G22 to C30; V23 to E31; R24 to P32; 125 to C33; V26 to G34; V27 to F35; E28 to E36; Y29 to A37; C30 to T38; E31 to Y39; P32 to L40; C33 to E41; G34 to L42; F35 to A43; E36 to S44; A37 to A45; T38 to V46; Y39 to K47;
TAO to E48; E41 to Q49; I42 to Y50; A43 to P51; S44 to G52; A45 to 153; V46 to E54; K47 to 155; E48 to E56; Q49 to S57; Y50 to R58; P51 to L59; G52 to G60; 153 to G61; E54 to T62; 155 to G63; E56 to A64; S57 to F65; R58 to E66;
L59 to 167; G60 to E68; G61 to 169; T62 to N70; G63 to G71; A64 to Q72; F65 to L73; E66 to V74;167 to F75; E68 to S76;169 to K77; N70 to L78; G71 to E79;
Q72 to N80; L73 to G81; V74 to G82; F75 to F83; S76 to P84; K77 to Y85; L78 to E86; E79 to K87; N80 to D88; G81 to L89; G82 to 190; F83 to E91; P84 to A92; Y85 to 193; E86 to R94; K87 to R95; D88 to A96; L89 to S97; 190 to N98;
E91 to G99; A92 to E100; 193 to T101; R94 to L102; R95 to E103; A96 to K104;
S97 to 1105; N98 to T106; G99 to N107; E100 to S108; T101 to R109; L102 to P110; E103 to P111; K104 to C112; 1105 to V113; T106 to 1114; and N107 to L115.
[0116] In a further embodiment, the isolated polypeptides of the present invention comprising or, alternatively, consisting of, one or more C35 peptide epitopes include the following lOmers (residues correspond to SEQ ID NO:2 and FIG. 1B): M1 to V10; S2 to All; G3 to P12; E4 to P13; P5 to P14; G6 to E15;
Q7 to E16; T8 to V17; S9 to E18; V10 to P19; All to G20; P12 to S21; P13 to G22; P14 to V23; El5 to R24; E16 to 125; V17 to V26; E18 to V27; P19 to E28;

G20 to Y29; S21 to C30; G22 to E31; V23 to P32; R24 to C33;125 to G34; V26 to F35; V27 to E36; E28 to A37; Y29 to T38; C30 to Y39; E31 to LAO; P32 to E41; C33 to L42; G34 to A43; F35 to S44; E36 to A45; A37 to V46; T38 to K47;
Y39 to E48; L40 to Q49; E41 to Y50; L42 to P51; A43 to G52; S44 to 153; A45 to E54; V46 to 155; K47 to E56; E48 to S57; Q49 to R58; Y50 to L59; P51 to G60; G52 to G61; 153 to T62; E54 to G63; 155 to A64; E56 to F65; S57 to E66;
R58 to 167; L59 to E68; G60 to 169; G61 to N70; T62 to G71; G63 to Q72; A64 to L73; F65 to V74; E66 to F75; 167 to S76; E68 to K77;169 to L78; N70 to E79;

G71 to N80; Q72 to G81; L73 to G82; V74 to F83; F75 to P84; S76 to Y85; K77 to E86; L78 to K87; E79 to D88; N80 to L89; G81 to 190; G82 to E91; F83 to A92; P84 to 193; Y85 to R94; E86 to R95; K87 to A96; D88 to S97; L89 to N98;
190 to G99; E91 to E100; A92 to T101; 193 to L102; R94 to E103; R95 to K104;
A96 to 1105; S97 to T106; N98 to N107; G99 to S108; E100 to R109; T101 to P110; L102 to P111; E103 to C112; K104 to V113;1105 to I114; T106 to L115.
[0117] In a further embodiment, the isolated polypeptides ofthe present invention comprising or, alternatively, consisting of, one or more C35 peptide epitopes include the following 1 lmers (residues correspond to SEQ ID NO:2 and FIG. 1B): M1 to All; S2 to P12; G3 to P13; E4 to P14; P5 to E15; G6 to E16;
Q7 to V17; T8 to E18; S9 to P19; V10 to G20; All to S21; P12 to G22; P13 to V23; P14 to R24; E15 to 125; E16 to V26; V17 to V27; E18 to E28; P19 to Y29;
G20 to C30; S21 to E31; G22 to P32; V23 to C33; R24 to G34; 125 to F35; V26 to E36; V27 to A37; E28 to T38; Y29 to Y39; C30 to 1240; E31 to E41; P32 to L42; C33 to A43; G34 to S44; F35 to A45; E36 to V46; A37 to K47; T38 to E48;
Y39 to Q49; IA0 to Y50; E41 to P51; L42 to G52; A43 to 153; S44 to E54; A45 to 155; V46 to E56; K47 to S57; E48 to R58; Q49 to L59; Y50 to G60; P51 to G61; G52 to T62; 153 to G63; E54 to A64; 155 to F65; E56 to E66; S57 to 167;
R58 to E68; L59 to 169; G60 to N70; G61 to G71; T62 to Q72; G63 to L73; A64 to V74; F65 to F75; E66 to S76;167 to K77; E68 to L78;169 to E79; N70 to N80;
G71 to G81; Q72 to G82; L73 to F83; V74 to P84; F75 to Y85; S76 to E86; K77 to K87; L78 to D88; E79 to L89; N80 to 190; G81 to E91; G82 to A92; F83 to 193; P84 to R94; Y85 to R95; E86 to A96; K87 to S97; D88 to N98; L89 to G99;
190 to E100; E91 to T101; A92 to L102; 193 to E103; R94 to K104; R95 to 1105;
A96 to T106; S97 to N107; N98 to S108; G99 to R109; E100 to P110; T101 to P111; L102 to C112; E103 to V113; K104 to 1114; 1105 to L115.
[0118] In another preferred embodiment, the isolated polypeptides of the present invention comprising or, alternatively, consisting of, C35 peptide epitopes include the following 12mers (residues correspond to SEQ ID NO:2 and FIG. 1B): M1 to P12; S2 to P13; G3 to P14; E4 to E15; P5 to E16; G6 to V17; Q7 to E18; T8 to P19; S9 to G20; V10 to S21; All to G22; P12 to V23; P13 to R24; P14 to 125;

E15 to V26; E16 to V27; V17 to E28; E18 to Y29; P19 to C30; G20 to E31; S21 to P32; G22 to C33; V23 to G34; R24 to F35; 125 to E36; V26 to A37; V27 to T38; E28 to Y39; Y29 to L40; C30 to E41; E31 to L42; P32 to A43; C33 to S44;
G34 to A45; F35 to V46; E36 to K47; A37 to E48; T38 to Q49; Y39 to Y50; L40 to P51; E41 to G52; L42 to 153; A43 to E54; S44 to 155; A45 to E56; V46 to S57;
K47 to R58; E48 to L59; Q49 to G60; Y50 to G61; P51 to T62; G52 to G63; 153 to A64; E54 to F65; 155 to E66; E56 to 167; S57 to E68; R58 to 169; L59 to N70;
G60 to G71; G61 to Q72; T62 to L73; G63 to V74; A64 to F75; F65 to S76; E66 to K77; 167 to L78; E68 to E79; 169 to N80; N70 to G81; G71 to G82; Q72 to F83; L73 to P84; V74 to Y85; F75 to E86; S76 to K87; K77 to D88; L78 to L89;
E79 to 190; N80 to E91; G81 to A92; G82 to 193; F83 to R94; P84 to R95; Y85 to A96; E86 to S97; K87 to N98; D88 to G99; L89 to E100; 190 to T101; E91 to L102; A92 to E103; 193 to K104; R94 to 1105; R95 to T106; A96 to N107; S97 to S108; N98 to R109; G99 to P110; E100 to P111; T101 to C112; L102 to V113; E103 to 1114; K104 to L115.
[0119] In another preferred embodiment, the isolated polypeptides of the present invention comprising or, alternatively, consisting of, one or more C35 peptide epitopes include the following 13mers (residues correspond to SEQ ID NO:2 and FIG. 1B): M1 to P13; S2 to P14; G3 to E15; E4 to E16; P5 to V17; G6 to E18;
Q7 to P19; T8 to G20; S9 to S21; V10 to G22; All to V23; P12 to R24; P13 to 125; P14 to V26; E15 to V27; E16 to E28; V17 to Y29; E18 to C30; P19 to E31;

G20 to P32; S21 to C33; G22 to G34; V23 to F35; R24 to E36; 125 to A37; V26 to T38; V27 to Y39; E28 to L40; Y29 to E41; C30 to L42; E31 to A43; P32 to S44; C33 to A45; G34 to V46; F35 to K47; E36 to E48; A37 to Q49; T38 to Y50;
Y39 to P51; L40 to G52; E41 to 153; L42 to E54; A43 to 155; S44 to E56; A45 to S57; V46 to R58; K47 to L59; E48 to G60; Q49 to G61; Y50 to T62; P51 to G63; G52 to A64; 153 to F65; E54 to E66; 155 to 167; E56 to E68; S57 to 169;
R58 to N70; L59 to G71; G60 to Q72; G61 to L73; T62 to V74; G63 to F75; A64 to S76; F65 to K77; E66 to L78; 167 to E79; E68 to N80; 169 to G81; N70 to G82; G71 to F83; Q72 to P84; L73 to Y85; V74 to E86; F75 to K87; S76 to D88;
K77 to L89; L78 to 190; E79 to E91; N80 to A92; G81 to 193; G82 to R94; F83 to R95; P84 to A96; Y85 to S97; E86 to N98; K87 to G99; D88 to E100; L89 to T101; 190 to L102; E91 to E103; A92 to K104; 193 to 1105; R94 to T106; R95 to N107; A96 to S108; S97 to R109; N98 to P110; G99 to P111; E100 to C112;
T101 to V113; L102 to 1114; E103 to L115.
[0120] In another preferred embodiment, the isolated polypeptides of the present invention comprising or, alternatively, consisting of, one or more C35 peptide epitopes include the following 14mers (residues correspond to SEQ ID NO:2 and FIG. 1B): M1 to P14; S2 to E15; G3 to E16; E4 to V17; P5 to E18; G6 to P19;
Q7 to G20; T8 to S21; S9 to G22; V10 to V23; All to R24; P12 to 125; P13 to V26; P14 to V27; E15 to E28; E16 to Y29; V17 to C30; E18 to E31; P19 to P32;
G20 to C33; S21 to G34; G22 to F35; V23 to E36; R24 to A37; 125 to T38; V26 to Y39; V27 to L40; E28 to E41; Y29 to L42; C30 to A43; E31 to S44; P32 to A45; C33 to V46; G34 to K47; F35 to E48; E36 to Q49; A37 to Y50; T38 to P51;
Y39 to G52; LAO to 153; E41 to E54; L42 to 155; A43 to E56; S44 to S57; A45 to R58; V46 to L59; K47 to G60; E48 to G61; Q49 to T62; Y50 to G63; P51 to A64; G52 to F65; 153 to E66; E54 to 167; 155 to E68; E56 to 169; S57 to N70;
R58 to G71; L59 to Q72; G60 to L73; G61 to V74; T62 to F75; G63 to S76; A64 to K77; F65 to L78; E66 to E79; 167 to N80; E68 to G81; 169 to G82; N70 to F83; G71 to P84; Q72 to Y85; L73 to E86; V74 to K87; F75 to D88; S76 to L89;
K77 to 190; L78 to E91; E79 to A92; N80 to 193; G81 to R94; G82 to R95; F83 to A96; P84 to S97; Y85 to N98; E86 to G99; K87 to E100; D88 to T101; L89 to L102; 190 to E103; E91 to K104; A92 to 1105; 193 to T106; R94 to N107; R95 to S108; A96 to R109; S97 to P110;N98 to P111; G99 to C112; E100 to V113;
T101 to 1114; L102 to L115.
[0121] In another preferred embodiment, the isolated polypeptides ofthe present invention comprising or, alternatively, consisting of, one or more C35 peptide epitopes include the following 15mers (residues correspond to SEQ ID NO:2 and FIG. 1B): M1 to E15; S2 to E16; G3 to V17; E4 to E18; P5 to P19; G6 to G20;
Q7 to S21; T8 to G22; S9 to V23; V10 to R24; All to 125; P12 to V26; P13 to V27; P14 to E28; El5 to Y29; E16 to C30; V17 to E31; El8 to P32; P19 to C33;
G20 to G34; S21 to F35; G22 to E36; V23 to A37; R24 to T38; 125 to Y39; V26 to L40; V27 to E41; E28 to L42; Y29 to A43; C30 to S44; E31 to A45; P32 to V46; C33 to K47; G34 to E48; F35 to Q49; E36 to Y50; A37 to P51; T38 to G52;
Y39 to 153; LAO to E54; E41 to 155; L42 to E56; A43 to S57; S44 to R58; A45 to L59; V46 to G60; K47 to G61; E48 to T62; Q49 to G63; Y50 to A64; P51 to F65; G52 to E66; 153 to 167; E54 to E68; 155 to 169; E56 to N70; S57 to G71;
R58 to Q72; L59 to L73; G60 to V74; G61 to F75; T62 to S76; G63 to K77; A64 to L78; F65 to E79; E66 to N80;167 to G81; E68 to G82; 169 to F83; N70 to P84;

G71 to Y85; Q72 to E86; L73 to K87; V74 to D88; F75 to L89; S76 to 190; K77 to E91; L78 to A92; E79 to 193; N80 to R94; G81 to R95; G82 to A96; F83 to S97; P84 to N98; Y85 to G99; E86 to E100; K87 to T101; D88 to L102; L89 to E103; 190 to K104; E91 to 1105; A92 to T106; 193 to N107; R94 to S108; R95 to R109; A96 to P110; S97 to P111; N98 to C112; G99 to V113; E100 to 1114;
T101 to L115.
[0122] In a further preferred embodiment,the isolated polyp eptides of the present invention comprising or, alternatively, consisting of, one or more C35 peptide epitopes include the following 16mers (residues correspond to SEQ ID NO:2 and FIG. 1B): M1 to E16; S2 to V17; G3 to E18; E4 to P19; P5 to G20; G6 to S21;
Q7 to G22; T8 to V23; S9 to R24; V10 to 125; All to V26; P12 to V27; P13 to E28; P14 to Y29; E15 to C30; E16 to E31; V17 to P32; E18 to C33; P19 to G34;

G20 to F35; S21 to E36; G22 to A37; V23 to T38; R24 to Y39; 125 to TAO; V26 to E41; V27 to L42; E28 to A43; Y29 to S44; C30 to A45; E31 to V46; P32 to K47; C33 to E48; G34 to Q49; F35 to Y50; E36 to P51; A37 to G52; T38 to 153;
Y39 to E54; L40 to 155; E41 to E56; L42 to S57; A43 to R58; S44 to L59; A45 to G60; V46 to G61; K47 to T62; E48 to G63; Q49 to A64; Y50 to F65; P51 to E66; G52 to 167; 153 to E68; E54 to 169; 155 to N70; E56 to G71; S57 to Q72;
R58 to L73; L59 to V74; G60 to F75; G61 to S76; T62 to K77; G63 to L78; A64 to E79; F65 to N80; E66 to G81; 167 to G82; E68 to F83; 169 to P84; N70 to Y85; G71 to E86; Q72 to K87; L73 to D88; V74 to L89; F75 to 190; S76 to E91;
K77 to A92; L78 to 193; E79 to R94; N80 to R95; G81 to A96; G82 to S97; F83 to N98; P84 to G99; Y85 to E100; E86 to T101; K87 to L102; D88 to E103; L89 to K104; 190 to 1105; E91 to T106; A92 to N107; 193 to S108; R94 to R109; R95 to P110; A96 to P111; S97 to C112; N98 to V113; G99 to 1114; E100 to L115.
[0123] In another preferred embodiment, the isolated polypeptides of the present invention comprising or, alternatively, consisting of, one or more C35 peptide epitopes include the following 17mers: M1 to V17; S2 to El 8; G3 to P19; E4 to G20; P5 to S21; G6 to G22; Q7 to V23; T8 to 1R24; S9 to 125; V10 to V26; All to V27; P12 to E28; P13 to Y29; P14 to C30; E15 to E31; El6 to P32; V17 to C33; E18 to G34; P19 to F35; G20 to E36; S21 to A37; G22 to T38; V23 to Y39;
R24 to L40; 125 to E41; V26 to L42; V27 to A43; E28 to S44; Y29 to A45; C30 to V46; E31 to K47; P32 to E48; C33 to Q49; G34 to Y50; F35 to P51; E36 to G52; A37 to 153; T38 to E54; Y39 to 155; L40 to E56; E41 to S57; 1A-2 to R58;
A43 to L59; S44 to G60; A45 to G61; V46 to T62; K47 to G63; E48 to A64; Q49 to F65; Y50 to E66; P51 to 167; G52 to E68; 153 to 169; E54 to N70; 155 to G71;
E56 to Q72; S57 to L73; R58 to V74; L59 to F75; G60 to S76; G61 to K77; T62 to L78; G63 to E79; A64 to N80; F65 to G81; E66 to G82; 167 to F83; E68 to P84; 169 to Y85; N70 to E86; G71 to K87; Q72 to D88; L73 to L89; V74 to 190;
F75 to E91; S76 to A92; K77 to 193; L78 to R94; E79 to R95; N80 to A96; G81 to S97; G82 to N98; F83 to G99; P84 to E100; Y85 to T101; E86 to L102; K87 to E103; D88 to K104; L89 to 1105; 190 to T106; E91 to N107; A92 to S108; 193 to R109; R94 to P110; R95 to P111; A96 to C112; S97 to V113; N98 to 1114;
G99 to L115.
[0124] In another preferred embodiment, the isolated polypeptides of the present invention comprising or, alternatively, consisting of, one or more C35 peptide epitopes include the following 18mers (residues correspond to SEQ ID NO:2 and FIG. 1B): M1 to E18; S2 to P19; G3 to G20; E4 to S21; P5 to G22; G6 to V23;
Q7 to R24; T8 to 125; S9 to V26; V10 to V27; All to E28; P12 to Y29; P13 to C30; P14 to E31; EIS to P32; E16 to C33; V17 to 034; E18 to F35; P19 to E36;
G20 to A37; S21 to T38; 022 to Y39; V23 to L40; R24 to E41; 125 to L42; V26 to A43; V27 to S44; E28 to A45; Y29 to V46; C30 to K47; E31 to E48; P32 to Q49; C33 to Y50; G34 to P51; F35 to G52; E36 to 153; A37 to E54; T38 to 155;
Y39 to E56; L40 to S57; E41 to R58; L42 to L59; A43 to 060; S44 to G61; A45 to T62; V46 to 063; K47 to A64; E48 to F65; Q49 to E66; Y50 to 167; P51 to E68; G52 to 169; 153 to N70; E54 to 071; 155 to Q72; E56 to L73; S57 to V74;
R58 to F75; L59 to S76; 060 to K77; G61 to L78; T62 to E79; G63 to N80; A64 to 081; F65 to 082; E66 to F83; 167 to P84; E68 to Y85; 169 to E86; N70 to K87; G71 to D88; Q72 to L89; L73 to 190; V74 to E91; F75 to A92; S76 to 193;
K77 to R94; L78 to R95; E79 to A96; N80 to S97; G81 to N98; 082 to G99; F83 to E100; P84 to T101; Y85 to L102; E86 to E103; K87 to K104; D88 to 1105;
L89 to T106;190 toN107;E91 to S108; A92 to R109; 193 toP110; R94 toP111;
R95 to C112; A96 to V113; S97 to 1114; N98 to L115.
[0125] In another preferred embodiment, the isolated polypeptides of the present invention comprising or, alternatively, consisting of, one or more C35 peptide epitopes include the following 19mers (residues correspond to SEQ ID NO:2 and FIG. 1B): M1 to P19; S2 to 020; G3 to S21; E4 to 022; P5 to V23; G6 to R24;
Q7 to 125; T8 to V26; S9 to V27; V10 to E28; All to Y29; P12 to C30; P13 to E31; P14 to P32; E15 to C33; E16 to G34; V17 to F35; El8 to E36; P19 to A37;
020 to T38; S21 to Y39; G22 to MO; V23 to E41; R24 to L42; 125 to A43; V26 to S44; V27 to A45; E28 to V46; Y29 to K47; C30 to E48; E31 to Q49; P32 to Y50; C33 to P51; G34 to 052; F35 to 153; E36 to E54; A37 to 155; T38 to E56;

Y39 to S57; L40 to R58; E41 to L59; L42 to G60; A43 to G61; S44 to T62; A45 to G63; V46 to A64; K47 to F65; E48 to E66; Q49 to 167; Y50 to E68; P51 to 169; G52 to N70; 153 to G71; E54 to Q72; 155 to L73; E56 to V74; S57 to F75;
R58 to S76; L59 to K77; G60 to L78; G61 to E79; T62 to N80; G63 to G81; A64 to G82; F65 to F83; E66 to P84; 167 to Y85; E68 to E86; 169 to K87; N70 to D88; G71 to L89; Q72 to 190; L73 to E91; V74 to A92; F75 to 193; S76 to R94;
K77 to R95; L78 to A96; E79 to S97; N80 to N98; G81 to G99; G82 to E100;
F83 to T101; P84 to L102; Y85 to E103; E86 to K104; K87 to 1105; D88 to T106; L89 to N107; 190 to S108; E91 to R109; A92 to P110; 193 to P111; R94 to C112; R95 to V113; A96 to 1114; S97 to L115.
[0126] In another preferred embodiment, the isolated polypeptides of the present invention comprising or, alternatively, consisting of, one or more C35 peptide epitopes include the following 20mers (residues correspond to SEQ ID NO:2 and FIG. 1B): M1 to G20; S2 to S21; G3 to G22; E4 to V23; P5 to R24; G6 to 125;
Q7 to V26; T8 to V27; S9 to E28; V10 to Y29; All to C30; P12 to E31; P13 to P32; P14 to C33; E15 to G34; E16 to F35; V17 to E36; E18 to A37; P19 to T38;
G20 to Y39; S21 to IA-0; G22 to E41; V23 to L42; R24 to A43; 125 to S44; V26 =
to A45; V27 to V46; E28 to K47; Y29 to E48; C30 to Q49; E31 to Y50; P32 to P51; C33 to G52; G34 to 153; F35 to E54; E36 to 155; A37 to E56; T38 to S57;
Y39 to R58; I.A.0 to L59; E41 to G60; L42 to G61; A43 to T62; S44 to G63; A45 to A64; V46 to F65; K47 to E66; E48 to 167; Q49 to E68; Y50 to 169; P51 to N70; G52 to G71; 153 to Q72; E54 to L73; 155 to V74; E56 to F75; S57 to S76;
R58 to K77; L59 to L78; G60 to E79; G61 to N80; T62 to G81; G63 to G82; A64 to F83; F65 to P84; E66 to Y85; 167 to E86; E68 to K87; 169 to D88; N70 to L89;
G71 to 190; Q72 to E91; L73 to A92; V74 to 193; F75 to R94; S76 to R95; K77 to A96; L78 to S97; E79 to N98; N80 to G99; G81 to E100; G82 to T101; F83 to L102; P84 to E103; Y85 to K104; E86 to 1105; K87 to T106; D88 to N107;
L89 to S108;190 to R109; E91 toP110; A92 toP111;193 to C112R94 to V113;
R95 to 1114; A96 to L115.

[0127] In another preferred embodiment, the isolated polypeptides of the present invention comprising or, alternatively, consisting of, one or more C35 peptide epitopes include the following 21mers (residues correspond to SEQ ID NO:2 and FIG. 1B): M1 to S21; S2 to G22; G3 to V23; E4 to R24; P5 to 125; G6 to V26;
Q7 to V27; T8 to E28; S9 to Y29; V10 to C30; All to E31; P12 to P32; P13 to C33; P14 to G34; E15 to F35; E16 to E36; V17 to A37; E18 to T38; P19 to Y39;
G20 to L40; S21 to E41; G22 to L42; V23 to A43; R24 to S44; 125 to A45; V26 to V46; V27 to K47; E28 to E48; Y29 to Q49; C30 to Y50; E31 to P51; P32 to G52; C33 to 153; G34 to E54; F35 to 155; E36 to E56; A37 to S57; T38 to R58;
Y39 to L59; L40 to G60; E41 to G61; L42 to T62; A43 to G63; S44 to A64; A45 to F65; V46 to E66; K47 to 167; E48 to E68; Q49 to 169; Y50 to N70; P51 to G71; G52 to Q72; 153 to L73; E54 to V74; 155 to F75; E56 to S76; S57 to K77;
R58 to L78; L59 to E79; G60 to N80; G61 to G81; T62 to G82; G63 to F83; A64 to P84; F65 to Y85; E66 to E86; 167 to K87; E68 to D88; 169 to L89; N70 to 190;
G71 to E91; Q72 to A92; L73 to 193; V74 to R94; F75 to R95; S76 to A96; K77 to S97; L78 to N98; E79 to G99; N80 to E100; G81 to T101; G82 to L102; F83 to E103; P84 to K104; Y85 to 1105; E86 to T106; K87 to N107; D88 to S108;
L89 to R109; I90 to P110; E91 to P111; A92 to C112;193 to V113; R94 to I114;
R95 to L115.
[0128] In another preferred embodiment, the isolated polypeptides of the present invention comprising or, alternatively, consisting of, one or more C35 peptide epitopes include the following 22mers (residues correspond to SEQ ID NO:2 and FIG. 1B): M1 to G22; S2 to V23; G3 to R24; E4 to 125; P5 to V26; G6 to V27;
Q7 to E28; T8 to Y29; S9 to C30; V10 to E31; All to P32; P12 to C33; P13 to G34; P14 to F35; E15 to E36; E16 to A37; V17 to T38; E18 to Y39; P19 to L40;
G20 to E41; S21 to 1/42; G22 to A43; V23 to S44; R24 to A45; 125 to V46; V26 to K47; V27 to E48; E28 to Q49; Y29 to Y50; C30 to P51; E31 to G52; P32 to 153; C33 to E54; G34 to 155; F35 to E56; E36 to S57; A37 to R58; T38 to L59;
Y39 to G60; L40 to G61; E41 to T62; L42 to G63; A43 to A64; S44 to F65; A45 to E66; V46 to 167; K47 to E68; E48 to 169; Q49 to N70; Y50 to G71; P51 to Q72; G52 to L73; 153 to V74; E54 to F75; 155 to S76; E56 to K77; S57 to L78;
R58 to E79; L59 to N80; G60 to G81; G61 to G82; T62 to F83; G63 to P84; A64 to Y85; F65 to E86; E66 to K87;167 to D88; E68 to L89; 169 to 190; N70 to E91;

G71 to A92; Q72 to 193; L73 to R94; V74 to R95; F75 to A96; S76 to S97; K77 to N98; L78 to G99; E79 to E100; N80 to T101; G81 to L102; G82 to E103; F83 to K104; P84 to 1105; Y85 to T106; E86 to N107; K87 to S108; D88 to R109;
L89 to P110; I90 to P111; E91 to C112; A92 to V113;193 to I114; R94 to L115.
[0129] In another preferred embodiment, the isolated polypeptides of the present invention comprising or, alternatively, consisting of, one or more C35 peptide epitopes include the following 23mers (residues correspond to SEQ ID NO:2 and FIG. 1B): M1 to V23; S2 to R24; G3 to 125; E4 to V26; P5 to V27; G6 to E28;
Q7 to Y29; T8 to C30; S9 to E31; V10 to P32; All to C33; P12 to G34; P13 to F35; P14 to E36; E15 to A37; E16 to T38; V17 to Y39; E18 to L40; P19 to E41;
G20 to L42; S21 to A43; G22 to S44; V23 to A45; R24 to V46; 125 to K47; V26 to E48; V27 to Q49; E28 to Y50; Y29 to P51; C30 to G52; E31 to 153; P32 to E54; C33 to 155; G34 to E56; F35 to S57; E36 to R58; A37 to L59; T38 to G60;
Y39 to G61; L40 to T62; E41 to G63; L42 to A64; A43 to F65; S44 to E66; A45 to 167; V46 to E68; K47 to 169; E48 to N70; Q49 to G71; Y50 to Q72; P51 to L73; G52 to V74; 153 to F75; E54 to S76; 155 to K77; E56 to L78; S57 to E79;
R58 to N80; L59 to G81; G60 to G82; G61 to F83; T62 to P84; G63 to Y85; A64 to E86; F65 to K87; E66 to D88;167 to L89; E68 to 190; 169 to E91; N70 to A92;

G71 to 193; Q72 to R94; L73 to R95; V74 to A96; F75 to S97; S76 to N98; K77 to G99; L78 to E100; E79 to T101; N80 to L102; G81 to E103; G82 to K104;
F83 to 1105; P84 to T106; Y85 to N107; E86 to S108; 1(87 to R109; D88 to P110; L89 to P111; 190 to C112; E91 to V113; A92 to 1114; 193 to L115.
[0130] In another preferred embodiment, the isolated polypeptides of the present invention comprising or, alternatively, consisting of, one or more C35 peptide epitopes include the following 24mers (residues correspond to SEQ ID NO:2 and FIG. 1B): M1 to R24; S2 to 125; G3 to V26; E4 to V27; P5 to E28; G6 to Y29;
Q7 to C30; T8 to E31; S9 to P32; V10 to C33; All to G34; P12 to F35; P13 to E36; P14 to A37; E15 to T38; E16 to Y39; V17 to L40; E18 to E41; P19 to L42;
020 to A43; S21 to S44; G22 to A45; V23 to V46; R24 to K47; 125 to E48; V26 to Q49; V27 to Y50; E28 to P51; Y29 to G52; C30 to 153; E31 to E54; P32 to 155; C33 to E56; 034 to S57; F35 to R58; E36 to L59; A37 to G60; T38 to G61;
Y39 to T62; L40 to G63; E41 to A64; L42 to F65; A43 to E66; S44 to 167; A45 to E68; V46 to 169; K47 to N70; E48 to G71; Q49 to Q72; Y50 to L73; P51 to V74; G52 to F75; 153 to S76; E54 to K77; 155 to L78; E56 to E79; S57 to N80;
R58 to G81; L59 to G82; 060 to F83; G61 to P84; T62 to Y85; G63 to E86; A64 to K87; F65 to D88; E66 to L89; 167 to 190; E68 to E91; 169 to A92; N70 to 193;
071 to R94; Q72 to R95; L73 to A96; V74 to S97; F75 to N98; S76 to G99; K77 to E100; L78 to T101; E79 to L102; N80 to E103; G81 to K104; G82 to 1105;
F83 to T106; P84 to N107; Y85 to S108; E86 to R109; K87 to P110; D88 to P111; L89 to C112; 190 to V113; E91 to 1114; A92 to L115.
[0131] In another preferred embodiment, the isolated polypeptides of the present invention comprising or, alternatively, consisting of, one or more C35 peptide epitopes include the following 25mers (residues correspond to SEQ ID NO:2 and FIG. 1B): M1 to 125; S2 to V26; G3 to V27; E4 to E28; P5 to Y29; G6 to C30;
Q7 to E31; T8 to P32; S9 to C33; V10 to G34; All to F35; P12 to E36; P13 to A37; P14 to T38; EIS to Y39; E16 to LAO; V17 to E41; EIS to L42; P19 to A43;
G20 to S44; S21 to A45; 022 to V46; V23 to K47; R24 to E48; 125 to Q49; V26 to Y50; V27 to P51; E28 to G52; Y29 to 153; C30 to E54; E31 to 155; P32 to E56; C33 to S57; G34 to R58; F35 to L59; E36 to G60; A37 to G61; T38 to T62;
Y39 to G63; L40 to A64; E41 to F65; L42 to E66; A43 to 167; S44 to E68; A45 to 169; V46 to N70; K47 to G71; E48 to Q72; Q49 to L73; Y50 to V74; P51 to F75; G52 to S76; 153 to K77; E54 to L78; 155 to E79; E56 to N80; S57 to G81;
R58 to G82; L59 to F83; G60 to P84; G61 to Y85; T62 to E86; G63 to K87; A64 to D88; F65 to L89; E66 to 190; 167 to E91; E68 to A92; 169 to 193; N70 to R94;
G71 to R95; Q72 to A96; L73 to S97; V74 to N98; F75 to G99; S76 to E100;
K77 to T101; L78 to L102; E79 to E103; N80 to K104; G81 to 1105; 082 to T106; F83 to N107; P84 to S108; Y85 to R109; E86 to P110; K87 to P111; D88 to C112; L89 to V113; 190 to 1114; E91 to L115.
[0132] In another preferred embodiment, the isolated polypeptides of the present invention comprising or, alternatively, consisting of, one or more C35 peptide epitopes include the following 26mers (residues correspond to SEQ ID NO:2 and FIG. 1B): M1 to V26; S2 to V27; G3 to E28; E4 to Y29; P5 to C30; G6 to E31;
Q7 to P32; T8 to C33; S9 to G34; V10 to F35; Al 1 to E36; P12 to A37; P13 to T38; P14 to Y39; EIS to L40; El6 to E41; V17 to L42; E18 to A43; P19 to S44;
G20 to A45; S21 to V46; G22 to K47; V23 to E48; R24 to Q49;125 to Y5O; V26 to P51; V27 to G52; E28 to 153; Y29 to E54; C30 to 155; E31 to E56; P32 to S57;
C33 to R58; G34 to L59; F35 to G60; E36 to G61; A37 to T62; T38 to G63; Y39 to A64; L40 to F65; E41 to E66; L42 to 167; A43 to E68; S44 to 169; A45 to N70;
V46 to 071; K47 to Q72; E48 to L73; Q49 to V74; Y50 to F75; P51 to S76; G52 to K77; 153 to L78; E54 to E79; 155 to N80; E56 to G81; S57 to G82; R58 to F83; L59 to P84; G60 to Y85; 061 to E86; T62 to K87; G63 to D88; A64 to L89;
F65 to 190; E66 to E91; 167 to A92; E68 to 193; 169 to R94; N70 to R95; 071 to A96; Q72 to S97; L73 to N98; V74 to 099; F75 to E100; S76 to T101; K77 to L102; L78 to E103; E79 to K104; N80 to 1105; G81 to T106; G82 to N107; F83 to S108; P84 to R109; Y85 to P110; E86 to P111; K87 to C112; D88 to V113;
L89 to 1114; 190 to L115.
[0133] In another preferred embodiment,the isolated polypeptides of the present invention comprising or, alternatively, consisting of, one or more C35 peptide epitopes include the following 27mers (residues correspond to SEQ ID NO:2 and FIG. 1B): M1 to V27; S2 to E28; G3 to Y29; E4 to C30; P5 to E31; G6 to P32;
Q7 to C33; T8 to G34; S9 to F35; V10 to E36; All to A37; P12 to T38; P13 to Y39; P14 to L40; E15 to E41; E16 to L42; V17 to A43; E18 to S44; P19 to A45;
G20 to V46; S21 to K47; G22 to E48; V23 to Q49; R24 to Y50; 125 to P51; V26 to 052; V27 to 153; E28 to E54; Y29 to 155; C30 to E56; E31 to S57; P32 to R58; C33 to L59; G34 to G60; F35 to G61; E36 to T62; A37 to 063; T38 to A64;
Y39 to F65; MO to E66; E41 to 167; L42 to E68; A43 to 169; S44 to N70; A45 to G71; V46 to Q72; K47 to L73; E48 to V74; Q49 to F75; Y50 to S76; P51 to K77; G52 to L78; 153 to E79; E54 to N80; 155 to G81; E56 to G82; S57 to F83;
R58 to P84; L59 to Y85; G60 to E86; G61 to K87; T62 to D88; G63 to L89; A64 to 190; F65 to E91; E66 to A92; 167 to 193; E68 to R94; 169 to R95; N70 to A96;
G71 to S97; Q72 to N98; L73 to G99; V74 to E100; F75 to T101; S76 to L102;
K77 to E103; L78 to K104; E79 to 1105; N80 to T106; G81 to N107; G82 to S108; F83 to R109; P84 to P110; Y85 to P111; E86 to C112; K87 to V113; D88 to 1114; L89 to L115.
[0134] In another preferred embodiment, the isolated polypeptides of the present invention comprising or, alternatively, consisting of, one or more C35 peptide epitopes include the following 28mers (residues correspond to SEQ ID NO:2 and FIG. 1B): M1 to E28; S2 to Y29; G3 to C30; E4 to E31; P5 to P32; G6 to C33;
Q7 to G34; T8 to F35; S9 to E36; V10 to A37; All to T38; P12 to Y39; P13 to L40; P14 to E41; E15 to L42; E16 to A43; V17 to S44; E18 to A45; P19 to V46;
G20 to K47; S21 to E48; G22 to Q49; V23 to Y50; R24 to P51; 125 to G52; V26 to 153; V27 to E54; E28 to 155; Y29 to E56; C30 to S57; E31 to R58; P32 to L59;
C33 to G60; G34 to G61; F35 to T62; E36 to G63; A37 to A64; T38 to F65; Y39 to E66; L40 to 167; E41 to E68; L42 to 169; A43 to N70; S44 to G71; A45 to " Q72; V46 to L73; K47 to V74; E48 to F75; Q49 to S76; Y50 to K77; P51 to L78;
G52 to E79; 153 to N80; E54 to G81; 155 to G82; E56 to F83; S57 to P84; R58 to Y85; L59 to E86; G60 to K87; G61 to D88; T62 to L89; G63 to 190; A64 to E91; F65 to A92; E66 to 193; 167 to R94; E68 to R95; 169 to A96; N70 to S97;
G71 to N98; Q72 to G99; L73 to E100; V74 to T101; F75 to L102; S76 to E103;
K77 to K104; L78 to 1105; E79 to T106; N80 to N107; G81 to S108; G82 to R109; F83 to P110; P84 to P111; Y85 to C112; E86 to V113; K87 to 1114; D88 to L115.
[0135] In another preferred embodiment, the isolated polypeptides of the present invention comprising or, alternatively, consisting of, one or more C35 peptide epitopes include the following 29mers (residues correspond to SEQ ID NO:2 and .
FIG. 1B): M1 to Y29; S2 to C30; G3 to E31; E4 to P32; P5 to C33; G6 to G34;

Q7 to F35; T8 to E36; S9 to A37; V10 to T38; All to Y39; P12 to L40; P13 to E41; P14 to L42; E15 to A43; E16 to S44; V17 to A45; E18 to V46; P19 to K47;
G20 to E48; S21 to Q49; G22 to Y50; V23 to P51; R24 to G52; 125 to 153; V26 to E54; V27 to 155; E28 to E56; Y29 to S57; C30 to R58; E31 to L59; P32 to G60; C33 to G61; G34 to T62; F35 to G63; E36 to A64; A37 to F65; T38 to E66;
Y39 to 167; L40 to E68; E41 to 169; L42 to N70; A43 to G71; S44 to Q72; A45 to L73; V46 to V74; K47 to F75; E48 to S76; Q49 to K77; Y50 to L78; P51 to E79; G52 to N80; 153 to G81; E54 to G82; 155 to F83; E56 to P84; S57 to Y85;
R58 to E86; L59 to K87; G60 to D88; G61 to L89; T62 to 190; G63 to E91; A64 to A92; F65 to 193; E66 to R94; 167 to R95; E68 to A96;169 to S97; N70 to N98;

G71 to G99; Q72 to E100; L73 to T101; V74 to L102; F75 to E103; S76 to K104;
K77 to 1105; L78 to T106; E79 to N107; N80 to S108; G81 to R109; G82 to P110; F83 to P111; P84 to C112; Y85 to V113; E86 to 1114; K87 to L115.
[0136] In another preferred embodiment, the isolated polypeptides of the present invention comprising or, alternatively, consisting of, one or more C35 peptide epitopes include the following 30mers (residues correspond to SEQ ID NO:2 and FIG. 1B): M1 to C30; S2 to E31; G3 to P32; E4 to C33; P5 to G34; G6 to F35;
Q7 to E36; T8 to A37; S9 to T38; V10 to Y39; All to L40; P12 to E41; P13 to L42; P14 to A43; E15 to S44; E16 to A45; V17 to V46; E18 to K47; P19 to E48;
G20 to Q49; S21 to Y50; G22 to P51; V23 to G52; R24 to 153; 125 to E54; V26 to 155; V27 to E56; E28 to S57; Y29 to R58; C30 to L59; E31 to G60; P32 to G61; C33 to T62; G34 to G63; F35 to A64; E36 to F65; A37 to E66; T38 to 167;
Y39 to E68; L40 to 169; E41 to N70; L42 to G71; A43 to Q72; S44 to L73; A45 to V74; V46 to F75; K47 to S76; E48 to K77; Q49 to L78; Y50 to E79; P51 to N80; G52 to G81; 153 to G82; E54 to F83; 155 to P84; E56 to Y85; S57 to E86;
R58 to K87; L59 to D88; G60 to L89; G61 to 190; T62 to E91; G63 to A92; A64 to 193; F65 to R94; E66 to R95;167 to A96; E68 to S97; 169 to N98; N70 to G99;
G71 to E100; Q72 to T101; L73 to L102; V74 to E103; F75 to K104; S76 to 1105; K77 to T106; L78 to N107; E79 to S108; N80 to R109; G81 to P110; G82 to P111; F83 to C112; P84 to V113; Y85 to 1114; E86 to L115.

[0137] In another preferred embodiment,the isolated polypeptides of the present invention comprising or, alternatively, consisting of, one or more C35 peptide epitopes include the following 3 lmers (residues correspond to SEQ ED NO:2 and FIG. 1B): M1 to E31; S2 to P32; G3 to C33; E4 to G34; P5 to F35; G6 to E36;
Q7 to A37; T8 to T38; S9 to Y39; V10 to MO; All to E41; P12 to L42; P13 to A43; P14 to S44; E15 to A45; E16 to V46; V17 to K47; El 8 to E48; P19 to Q49;
G20 to Y50; S21 to P51; G22 to G52; V23 to 153; R24 to E54; 125 to 155; V26 to E56; V27 to S57; E28 to R58; Y29 to L59; C30 to G60; E31 to G61; P32 to T62; C33 to G63; G34 to A64; F35 to F65; E36 to E66; A37 to 167; T38 to E68;
Y39 to 169; MO to N70; E41 to G71; L42 to Q72; A43 to L73; S44 to V74; A45 .to F75; V46 to S76; K47 to K77; E48 to L78; Q49 to E79; Y50 to N80; P51 to G81; G52 to G82; 153 to F83; E54 to P84;155 to Y85; E56 to E86; S57 to K87;
R58 to D88; L59 to L89; G60 to 190; G61 to E91; T62 to A92; G63 to 193; A64 to R94; F65 to R95; E66 to A96; 167 to S97; E68 to N98; 169 to G99; N70 to E100; G71 to T101; Q72 to L102; L73 to E103; V74 to K104; F75 to 1105; S76 to T106; K77 to N107; L78 to S108; E79 to R109; N80 to P110; G81 to P111;
G82 to C112; F83 to V113; P84 to 1114 and Y85 to L115.
[0138] In another preferred embodiment,the isolated polypeptides of the present invention comprising or, alternatively, consisting of, one or more C35 peptide epitopes include the following 32mers (residues correspond to SEQ ID NO:2 and FIG. 1B): M1 to P32; S2 to C33; G3 to G34; E4 to F35; P5 to E36; G6 to A37;
Q7 to T38; T8 to Y39; S9 to MO; V10 to E41; All to L42; P12 to A43; P13 to S44; P14 to A45; EIS to V46; E16 to K47; V17 to E48; El8 to Q49; P19 to Y50;
G20 to P51; S21 to G52; G22 to 153; V23 to E54; R24 to 155; 125 to E56; V26 to S57; V27 to R58; E28 to L59; Y29 to G60; C30 to G61; E31 to T62; P32 to G63; C33 to A64; G34 to F65; F35 to E66; E36 to 167; A37 to E68; T38 to 169;
Y39 to N70; MO to G71; E41 to Q72; L42 to L73; A43 to V74; S44 to F75; A45 to S76; V46 to K77; K47 to L78; E48 to E79; Q49 to N80; Y50 to G81; P51 to G82; G52 to F83; 153 to P84; E54 to Y85; 155 to E86; E56 to K87; S57 to D88;
R58 to L89; L59 to 190; G60 to E91; G61 to A92; T62 to 193; G63 to R94; A64 to R95; F65 to A96; E66 to S97; 167 to N98; E68 to G99; 169 to E100; N70 to T101; G71 to L102; Q72 to E103; L73 to K104; V74 to 1105; F75 to T106; S76 to N107; K77 to S108; L78 to R109; E79 to P110; N80 to P111; G81 to C112;
G82 to V113; F83 to 1114 and P84 to L115.
[0139] In another preferred embodiment,the isolated polypeptides of the present invention comprising or, alternatively, consisting of, one or more C35 peptide epitopes include the following 33mers (residues correspond to SEQ ID NO:2 and FIG. 1B): M1 to C33; S2 to G34; G3 to F35; E4 to E36; P5 to A37; G6 to T38;
Q7 to Y39; T8 to L40; S9 to E41; V10 to L42; All to A43; P12 to S44; P13 to A45; P14 to V46; E15 to K47; E16 to E48; V17 to Q49; E18 to Y50; P19 to P51;
G20 to G52; S21 to 153; G22 to E54; V23 to 155; R24 to E56; 125 to S57; V26 to R58; V27 to L59; E28 to G60; Y29 to G61; C30 to T62; E31 to G63; P32 to A64; C33 to F65; G34 to E66; F35 to 167; E36 to E68; A37 to 169; T38 to N70;
Y39 to G71; TAO to Q72; E41 to L73; L42 to V74; A43 to F75; S44 to S76; A45 to K77; V46 to L78; K47 to E79; E48 to N80; Q49 to G81; Y50 to G82; P51 to F83; G52 to P84; 153 to Y85; E54 to E86; 155 to K87; E56 to D88; S57 to L89;
R58 to 190; L59 to E91; G60 to A92; G61 to 193; T62 to R94; G63 to R95; A64 to A96; F65 to S97; E66 to N98; 167 to G99; E68 to E100; 169 to T101; N70 to L102; G71 to E103; Q72 to K104; L73 to 1105; V74 to T106; F75 to N107; S76 to S108; K77 to R109; L78 to P110; E79 to P111; N80 to C112; G81 to V113;
G82 to 1114 and F83 to L115.
[0140] In another preferred embodiment,the isolated polypeptides of the present invention comprising or, alternatively, consisting of, one or more C35 peptide epitopes include the following 34mers (residues correspond to SEQ ID NO:2 and FIG. 1B): M1 to G34; S2 to F35; G3 to E36; E4 to A37; P5 to T38; G6 to Y39;
Q7 to L40; T8 to E41; S9 to L42; V10 to A43; All to S44; P12 to A45; P13 to V46; P14 to K47; EIS to E48; E16 to Q49; V17 to Y50; EIS to P51; P19 to G52;
G20 to 153; S21 to E54; G22 to 155; V23 to E56; R24 to S57; 125 to R58; V26 to L59; V27 to G60; E28 to G61; Y29 to T62; C30 to G63; E31 to A64; P32 to F65;
C33 to E66; G34 to 167; F35 to E68; E36 to 169; A37 to N70; T38 to G71; Y39 to Q72; L40 to L73; E41 to V74; L42 to F75; A43 to S76; S44 to K77; A45 to L78; V46 to E79; K47 to N80; E48 to G81; Q49 to G82; Y50 to F83; P51 to P84;
G52 to Y85; 153 to E86; E54 to K87; 155 to D88; E56 to L89; S57 to 190; R58 to E91; L59 to A92; G60 to 193; G61 to R94; T62 to R95; G63 to A96; A64 to S97;
F65 to N98; E66 to G99; 167 to E100; E68 to T101; 169 to L102; N70 to E103;
G71 to K104; Q72 to 1105; L73 to T106; V74 to N107; F75 to S108; S76 to R109; K77 to P110; L78 to P111; E79 to C112; N80 to V113; G81 to 1114 and G82 to L115.
[0141] In another preferred embodiment,the isolated polypeptides of the present invention comprising or, alternatively, consisting of, one or more C35 peptide epitopes include the following 35mers (residues correspond to SEQ ID NO:2 and FIG. 1B): M1 to F35; S2 to E36; G3 to A37; E4 to T38; P5 to Y39; G6 to L40;
Q7 to E41; T8 to L42; S9 to A43; V10 to S44; All to A45; P12 to V46; P13 to K47; P14 to E48; E15 to Q49; E16 to Y50; V17 to P51; E18 to G52; P19 to 153;
G20 to E54; S21 to 155; G22 to E56; V23 to S57; R24 to R58; 125 to L59; V26 to G60; V27 to G61; E28 to T62; Y29 to G63; C30 to A64; E31 to F65; P32 to E66; C33 to 167; G34 to E68; F35 to 169; E36 to N70; A37 to G71; T38 to Q72;
Y39 to L73; 1,40 to V74; E41 to F75; 142 to S76; A43 to K77; S44 to L78; A45 to E79; V46 to N80; K47 to G81; E48 to G82; Q49 to F83; Y50 to P84; P51 to Y85; G52 to E86; 153 to K87; E54 to D88; 155 to L89; E56 to 190; S57 to E91;
R58 to A92; L59 to 193; G60 to R94; G61 to R95; T62 to A96; G63 to S97; A64 to N98; F65 to G99; E66 to E100; 167 to T101; E68 to L102; 169 to E103; N70 to K104; G71 to 1105; Q72 to T106; L73 to N107; V74 to S108; F75 to R109;
S76 to P110; K77 to P111; L78 to C112; E79 to V113; N80 to 1114; G81 to L115.
[0142] In another preferred embodiment, the isolated polypeptides of the present invention comprising or, alternatively, consisting of, one or more C35 peptide epitopes include the following 36mers (residues correspond to SEQ ID NO:2 and FIG. 1B):

M1 to E36; S2 to A37; G3 to T38; E4 to Y39; P5 to L40; G6 to E41; Q7 to L42;
T8 to A43; S9 to S44; V10 to A45; All to V46; P12 to K47; P13 to E48; P14 to Q49; E15 to Y50; E16 to P51; V17 to G52; El8 to 153; P19 to E54; G20 to 155;
S21 to E56; G22 to S57; V23 to R58; R24 to L59; 125 to G60; V26 to G61; V27 to T62; E28 to G63; Y29 to A64; C30 to F65; E31 to E66; P32 to 167; C33 to E68; G34 to 169; F35 to N70; E36 to G71; A37 to Q72; T38 to L73; Y39 to V74;
L40 to F75; E41 to S76; L42 to K77; A43 to L78; S44 to E79; A45 to N80; V46 to G81; K47 to G82; E48 to F83; Q49 to P84; Y50 to Y85; P51 to E86; G52 to K87; 153 to D88; E54 to L89; 155 to 190; E56 to E91; S57 to A92; R58 to 193;
L59 to R94; G60 to R95; G61 to A96; T62 to S97; G63 to N98; A64 to G99; F65 to E100; E66 to T101;167 to L102; E68 to E103; 169 to K104; N70 to Il05; G71 to T106; Q72 to N107; L73 to S108; V74 to R109; F75 to P110; S76 to P111;
K77 to C112; L78 to V113; E79 to 1114; N80 to L115.
[0143] In another preferred embodiment, the isolated polypeptides of the present invention comprising or, alternatively, consisting of, one or more C35 peptide epitopes include the following 37mers (residues correspond to SEQ ID NO:2 and FIG. 1B):
M1 to A37; S2 to T38; G3 to Y39; E4 to L40; P5 to E41; G6 to L42; Q7 to A43;
T8 to S44; S9 to A45; V10 to V46; All to K47; P12 to B48; P13 to Q49; P14 to Y50; E15 to P51; E16 to G52; V17 to 153; E18 to E54; P19 to 155; G20 to E56;
S21 to S57; G22 to R58; V23 to L59; R24 to G60; 125 to G61; V26 to T62; V27 to G63; E28 to A64; Y29 to F65; C30 to E66; E31 to 167; P32 to E68; C33 to 169; G34 to N70; F35 to G71; A37 to L73; T38 to V74; Y39 to F75; LAO to S76;
E41 to K77; L42 to L78; A43 to E79; S44 to N80; A45 to G81; V46 to G82; K47 to F83; E48 to P84; Q49 to Y85; Y50 to E86; P51 to K87; G52 to D88; 153 to L89; E54 to 190; 155 to E91; E56 to A92; S57 to 193; R58 to R94; L59 to R95;
G60 to A96; G61 to S97; T62 to N98; G63 to G99; A64 to E100; F65 to T101;
E66 to L102; 167 to E103; E68 to K104;169 to 1105; N70 to T106; G71 to N107;
Q72 to S108; L73 to R109; V74 to P110; F75 to P111; S76 to C112; K77 to V113; L78 to 1114; E79 to L115.

[0144] In another preferred embodiment, the isolated polypeptides of the present invention comprising or, alternatively, consisting of, one or more C35 peptide epitopes include the following 38mers (residues correspond to SEQ ID NO:2 and FIG. 1B):
M1 to T38; S2 to Y39; G3 to TAO; E4 to E41; P5 to L42; G6 to A43; Q7 to S44;
T8 to A45; S9 to V46; V10 to K47; All to E48; P12 to Q49; P13 to Y50; P14 to P51; El5 to G52; El6 to 153; V17 to E54; E18 to 155; P19 to E56; G20 to S57;
S21 to R58; G22 to L59; V23 to G60; R24 to G61;125 to T62; V26 to G63; V27 to A64; E28 to F65; Y29 to E66; C30 to 167; E31 to E68; P32 to 169; C33 to N70; G34 to G71; F35 to Q72; E36 to L73; A37 to V74; T38 to F75; Y39 to S76;
L40 to K77; E41 to L78; L42 to E79; A43 to N80; S44 to G81; A45 to G82; V46 to F83; K47 to P84; E48 to Y85; Q49 to E86; Y50 to K87; P51 to D88; G52 to L89; 153 to 190; E54 to E91; 155 to A92; E56 to 193; S57 to R94; R58 to R95;
L59 to A96; G60 to S97; G61 to N98; T62 to G99; G63 to E100; A64 to T101;
F65 to L102; E66 to E103; 167 to K104; E68 to 1105; 169 to T106; N70 to N107;
G71 to S108; Q72 to R109; L73 to P110; V74 to P111; F75 to C112; S76 to V113; K77 to 1114; L78 to L115.
[0145] In another preferred embodiment, the isolated polypeptides of the present invention comprising or, alternatively, consisting of, one or more C35 peptide epitopes include the following 39mers (residues correspond to SEQ ID NO:2 and FIG. 1B):
M1 to Y39; S2 to TAO; G3 to E41; E4 to L42; P5 to A43; G6 to S44; Q7 to A45;
T8 to V46; S9 to K47; V10 to E48; All to Q49; P12 to Y50; P13 to P51; P14 to G52; El5 to 153; E16 to E54; V17 to 155; El8 to E56; P19 to S57; G20 to R58;
S21 to L59; G22 to G60; V23 to G61; R24 to T62; 125 to G63; V26 to A64; V27 to F65; E28 to E66; Y29 to 167; C30 to E68; E31 to 169; P32 to N70; C33 to G71; G34 to Q72; F35 to L73; E36 to V74; A37 to F75; T38 to S76; Y39 to K77;
TAO to L78; E41 to E79; L42 to N80; A43 to G81; S44 to G82; A45 to F83; V46 to P84; K47 to Y85; E48 to E86; Q49 to K87; Y50 to D88; P51 to L89; G52 to 190; 153 to E91; E54 to A92; 155 to 193; E56 to R94; S57 to R95; R58 to A96;

L59 to S97; G60 to N98; G61 to G99; T62 to E100; G63 to T101; A64 to L102;
F65 to E103; E66 to K104; 167 to 1105; E68 to T106; 169 to N107; N70 to S108;
G71 to R109; Q72 to P110; L73 to P111; V74 to C112; F75 to V113; S76 to 1114; K77 to L115.
[0146] In another preferred embodiment, the isolated polypeptides of the present invention comprising or, alternatively, consisting of, one or more C35 peptide epitopes include the following 40mers (residues correspond to SEQ ID NO:2 and FIG. 1B):
M1 to MO; S2 to E41; G3 to L42; E4 to A43; P5 to S44; G6 to A45; Q7 to V46;
T8 to K47; S9 to E48; V10 to Q49; All to Y50; P12 to P51; P13 to G52; P14 to 153; E15 to E54; E16 to 155; V17 to E56; E18 to S57; P19 to R58; G20 to L59;
S21 to G60; G22 to G61; V23 to T62; R24 to G63; 125 to A64; V26 to F65; V27 to E66; E28 to 167; Y29 to E68; C30 to 169; E31 to N70; P32 to G71; C33 to Q72; G34 to L73; F35 to V74; E36 to F75; A37 to S76; T38 to K77; Y39 to L78;
L40 to E79; E41 to N80; IA2 to G81; A43 to G82; S44 to F83; A45 to P84; V46 to Y85; K47 to E86; E48 to K87; Q49 to D88; Y50 to L89; P51 to 190; G52 to E91; 153 to A92; E54 to 193; 155 to R94; E56 to R95; S57 to A96; R58 to S97;
L59 to N98; G60 to G99; G61 to E100; T62 to T101; G63 to L102; A64 to E103;
F65 to K104; E66 to 1105; 167 to T106; E68 to N107; 169 to S108; N70 to R109;
G71 to P110; Q72 to P111; L73 to C112; V74 to V113; F75 to 1114; S76 to L115.
[0147] In another preferred embodiment, the isolated polypeptides of the present invention comprising or, alternatively, consisting of, one or more C35 peptide epitopes include the following 41mers (residues correspond to SEQ lD NO :2 and FIG. 1B):
M1 to E41; S2 to L42; G3 to A43; E4 to S44; P5 to A45; G6 to V46; Q7 to K47;
T8 to E48; S9 to Q49; V10 to Y50; All to P51; P12 to G52; P13 to 153; P14 to E54; E15 to 155; E16 to E56; V17 to S57; El 8 to R58; P19 to L59; G20 to G60;
S21 to G61; G22 to T62; V23 to G63; R24 to A64; 125 to F65; V26 to E66; V27 to 167; E28 to E68;Y29 to 169; C30 to N70; E31 to G71; P32 to Q72; C33 to L73;

G34 to V74; F35 to F75; E36 to S76; A37 to K77; T38 to L78; Y39 to E79; MO
to N80; E41 to G81; L42 to G82; A43 to F83; S44 to P84; A45 to Y85; V46 to E86; K47 to K87; E48 to D88; Q49 to L89; Y50 to 190; P51 to E91; G52 to A92;
153 to 193; E54 to R94; 155 to R95; E56 to A92; S57 to S97; R58 to N98; L59 to G99; G60 to E100; G61 to T101; T62 to L102; G63 to E103; A64 to K104; F65 to 1105; E66 to T106; 167 to N107; E68 to S108; 169 to R109; N70 to P110; G71 to P111; Q72 to C112; L73 to V113; V74 to 1114; F75 to L115.
101481 In another preferred embodiment, the isolated polypeptides of the present invention comprising or, alternatively, consisting of, one or more C35 peptide epitopes include the following 42mers (residues correspond to SEQ ID NO:2 and FIG. 1B):
M1 to L42; S2 to A43; G3 to S44; E4 to A45; P5 to V46; G6 to K47; Q7 to E48;
T8 to Q49; S9 to Y50; V10 to P51; All to G52; P12 to 153; P13 to E54; P14 to 155; E15 to E56; E16 to S57; V17 to R58; E18 to L59; P19 to G60; G20 to G61;
S21 to T62; G22 to G63; V23 to A64; R24 to F65; 125 to E66; V26 to 167; V27 to E68; E28 to 169; Y29 to N70; C30 to G71; E31 to Q72; P32 to L73; C33 to V74; G34 to F75; F35 to S76; E36 to K77; A37 to L78; T38 to E79; Y39 to N80;
1_,40 to G81; E41 to G82; L42 to F83; A43 to P84; S44 to Y85; A45 to E86; V46 to K87; K47 to D88; E48 to L89; Q49 to 190; Y50 to E91; P51 to A92; G52 to 193; 153 to R94; E54 to R95; 155 to A96; E56 to S97; S57 to N98; R58 to G99;
L59 to E100; G60 to T101; G61 to L102; T62 to E103; G63 to K104; A64 to 1105; F65 to T106; E66 to N107; 167 to S108; E68 to R109; 169 to P110; N70 to P111; G71 to C112; Q72 to V113; L73 to 1114; V74 to L115.
[0149] In another preferred embodiment, the isolated polypeptides of the present invention comprising or, alternatively, consisting of, one or more C35 peptide epitopes include the following 43mers (residues correspond to SEQ ID NO:2 and FIG. 1B):
M1 to A43; S2 to S44; G3 to A45; E4 to V46; P5 to K47; G6 to E48; Q7 to Q49;
T8 to Y50; S9 to P51; V10 to G52; All to 153; P12 to E54; P13 to 155; P14 to E56; E15 to S57; El6 to R58; V17 to L59; E18 to G60; P19 to G61; G20 to T62;

S21 to G63; G22 to A64; V23 to F65; R24 to E66; 125 to 167; V26 to E68; V27 to 169; E28 to N70; Y29 to G71; C30 to Q72; E31 to L73; P32 to V74; C33 to F75; 034 to S76; F35 to K77; E36 to L78; A37 to E79; T38 to N80; Y39 to G81;
L40 to G82; E41 to F83; L42 to P84; A43 to Y85; S44 to E86; A45 to K87; V46 to D88; K47 to L89; E48 to 190; Q49 to E91; Y50 to A92; P51 to 193; 052 to R94; 153 to R95; E54 to A96; 155 to S97; E56 to N98; S57 to G99; R58 to E100;
L59 to T101; 060 to L102; G61 to E103; T62 to K104; 063 to 1105; A64 to T106; F65 to N107; E66 to S108; 167 to R109; E68 to P110; 169 to P111; N70 to C112; 071 to V113; Q72 to 1114; L73 to L115.
[0150] In another preferred embodiment, the isolated polypeptides of the present invention comprising or, alternatively, consisting of, one or more C35 peptide epitopes include the following 44mers (residues correspond to SEQ ID NO:2 and FIG. 1B):
M1 to S44; S2 to A45; G3 to V46; E4 to K47; P5 to E48; G6 to Q49; Q7 to Y50;
T8 to P51; S9 to G52; V10 to 153; All to E54; P12 to 155; P13 to E56; P14 to S57; E15 to R58; E16 to L59; V17 to G60; E18 to G61; P19 to T62; G20 to G63;
S21 to A64; G22 to F65; V23 to E66; R24 to 167; 125 to E68; V26 to 169; V27 to N70; E28 to G71; Y29 to Q72; C30 to L73; E31 to V74; P32 to F75; C33 to S76; G34 to K77; F35 to L78; E36 to E79; A37 to N80; T38 to G81; Y39 to G82;
L40 to F83; E41 to P84; L42 to Y85; A43 to E86; S44 to K87; A45 to D88; V46 to L89; K47 to 190; E48 to E91; Q49 to A92; Y50 to 193; P51 to R94; G52 to R95; 153 to A96; E54 to S97; 155 to N98; E56 to G99; S57 to E100; R58 to T101;

L59 to L102; G60 to E103; G61 to K104; T62 to 1105; G63 to T106; A64 to N107; F65 to S108; E66 to R109; 167 to P110; E68 to P111; 169 to C112; N70 to V113; 071 to 1114; Q72 to L115.
[0151] In another preferred embodiment, the isolated polypeptides of the present invention comprising or, alternatively, consisting of, one or more C35 peptide epitopes include the following 45mers (residues correspond to SEQ ID NO:2 and FIG. 1B):

M1 to A45; S2 to V46; G3 to K47; E4 to E48; P5 to Q49; G6 to Y50; Q7 to P51;
T8 to G52; S9 to 153; V10 to E54; All to 155; P12 to E56; P13 to S57; P14 to R58; E15 to L59; El 6 to G60; V17 to G61; E18 to T62; P19 to G63; G20 to A64;
S21 to F65; G22 to E66; V23 to 167; R24 to E68; 125 to 169; V26 to N70; V27.
to G71; E28 to Q72; Y29 to L73; C30 to V74; E31 to F75; P32 to S76; C33 to K77; G34 to L78; F35 to E79; E36 to N80; A37 to G81; T38 to G82; Y39 to F83;
L40 to P84; E41 to Y85; L42 to E86; A43 to K87; S44 to D88; A45 to L89; V46 to 190; K47 to E91; E48 to E92; Q49 to 193; Y50 to R94; P51 to R95; G52 to A96; 153 to S97; E54 to N98; 155 to G99; E56 to E100; S57 to T101; R58 to L102; L59 to E103; G60 to K104; G61 to K105; T62 to T106; G63 to N107; A64 to S108; F65 to R109; E66 to P110;167 to P111; E68 to C112;169 to V113; N70 to 1114; G71 to L115.
[0152] In another preferred embodiment, the isolated polypeptides of the present invention comprising or, alternatively, consisting of, one or more C35 peptide epitopes include the following 46mers (residues correspond to SEQ ID NO:2 and FIG. 1B):
M1 to V46; S2 to K47; G3 to E48; E4 to Q49; P5 to Y50; G6 to P51; Q7 to G52;
T8 to 153; S9 to E54; V10 to 155; All to E56; P12 to S57; P13 to R58; P14 to L59; E15 to G60; E16 to G61; V17 to T62; E18 to G63; P19 to A64; G20 to F65;
S21 to E66; G22 to 167; V23 to E68; R24 to 169; 125 to N70; V26 to G71; V27 to Q72; E28 to L73; Y29 to V74; C30 to F75; E31 to S76; P32 to K77; C33 to L78; G34 to E79; F35 to N80; E36 to G81; A37 to G82; T38 to F83; Y39 to P84;
MO to Y85; E41 to E86; L42 to K87; A43 to D88; S44 to L89; A45 to 190; V46 to E91; K47 to A92; E48 to 193; Q49 to R94; Y50 to R95; P51 to A96; G52 to S97; 153 to N98; E54 to G99; 155 to E100; E56 to T101; S57 to L102; R58 to E103; L59 to K104; G60 to 1105; G61 to T106; T62 to N107; G63 to S108; A64 to R109; F65 to P110; E66 to P111; 167 to C112; E68 to V113; 169 to 1114; N70 to L115 [0153] In another preferred embodiment, the isolated polypeptides of the present invention comprising or, alternatively, consisting of, one or more C35 peptide epitopes include the following 47mers (residues correspond to SEQ ID NO:2 and FIG. 1B):
M1 to K47; S2 to E48; G3 to Q49; E4 to Y50; P5 to P51; G6 to G52; Q7 to 153;
T8 to E54; S9 to 155; V10 to E56; All to S57; P12 to R58; P13 to L59; P14 to G60; El5 to G61; E16 to T62; V17 to G63; E18 to A64; P19 to F65; G20 to E66;
S21 to 167; G22 to E68; V23 to 169; R24 to N70; 125 to G71; V26 to Q72; V27 to L73; E28 to V74; Y29 to F75; C30 to S76; E31 to K77; P32 to L78; C33 to E79; G34 to N80; F35 to G81; E36 to G82; A37 to F83; T38 to P84; Y39 to Y85;
L40 to E86; E41 to K87; L42 to D88; A43 to L89; S44 to 190; A45 to E91; V46 to A92; K47 to 193; E48 to R94; Q49 to R95; Y50 to A96; P51 to S97; G52 to N98; 153 to G99; E54 to E100; 155 to T101; E56 to L102; S57 to E103; R58 to K104; L59 to 1105; G60 to T106; G61 to N107; T62 to S108; G63 to R109; A64 to P110; F65 to P111; E66 to C112; 167 to V113; E68 to 1114; 169 to L115 [0154] In another preferred embodiment, the isolated polypeptides of the present invention comprising or, alternatively, consisting of, one or more C35 peptide epitopes include the following 48mers (residues correspond to SEQ ID NO:2 and FIG. 1B):
M1 to E48; S2 to Q49; G3 to Y50; E4 to P51; P5 to G52; G6 to 153; Q7 to E54;
T8 to 155; S9 to E56; V10 to S57; All to R58; P12 to L59; P13 to G60; P14 to G61; E15 to T62; E16 to G63; V17 to A64; E18 to F65; P19 to E66; G20 to 167;
S21 to E68; G22 to 169; V23 to N70; R24 to G71; 125 to Q72; V26 to L73; V27 to V74; E28 to F75; Y29 to S76; C30 to K77; E31 to L78; P32 to E79; C33 to N80; G34 to G81; F35 to G82; E36 to F83; A37 to P84; T38 to Y85; Y39 to E86;
L40 to K87; E41 to D88; L42 to L89; A43 to 190; S44 to E91; A45 to A92; V46 to 193; K47 to R94; E48 to R95; Q49 to A96; Y50 to S97; P51 to N98; G52 to G99; 153 to E100; E54 to T101; 155 to L102; E56 to E103; S57 to K104; R58 to 1105; L59 to T106; G60 to N107; G61 to S108; T62 to R109; G63 to P110; A64 to P111; F65 to C112; E66 to V113; 167 to 1114; E68 to L115 [0155] In another preferred embodiment, the isolated polypeptides of the present invention comprising or, alternatively, consisting of, one or more C35 peptide epitopes include the following 49mers (residues correspond to SEQ ID NO:2 and FIG. 1B):
M1 to Q49; S2 to Y50; G3 to P51; E4 to G52; P5 to 153; G6 to E54; Q7 to 155;
T8 to E56; S9 to S57; V10 to R58; All to L59; P12 to G60; P13 to G61; P14 to T62; E15 to G63; E16 to A64; V17 to F65; E18 to E66; P19 to 167; G20 to E68;
S21 to 169; G22 to N70; V23 to G71; R24 to Q72; 125 to L73; V26 to V74; V27 to F75; E28 to S76; Y29 to K77; C30 to L78; E31 to E79; P32 to N80; C33 to G81; G34 to G82; F35 to F83; E36 to P84; A37 to Y85; T38 to E86; Y39 to K87;
L40 to D88; E41 to L89; L42 to 190; A43 to E91; S44 to A92; A45 to 193; V46 to R94; K47 to R95; E48 to A96; Q49 to S97; Y50 to N98; P51 to G99; G52 to E100; 153 to T101; E54 to L102; 155 to E103; E56 to K104; S57 to 1105; R58 to T106; L59 to N107; G60 to S108; G61 to R109; T62 to P110; G63 to P111; A64 to C112; F65 to V113; E66 to 1114; 167 to L115 [01561 In another preferred embodiment, the isolated polyp eptides of the present invention comprising or, alternatively, consisting of, one or more C35 peptide epitopes include the following 50mers (residues correspond to SEQ ID NO:2 and FIG. 1B):
M1 to Y50; S2 to P51; G3 to G52; E4 to 153;P5 to E54; G6 to 155; Q7 to E56;
T8 to S57; S9 to R58; V10 to L59; All to G60; P12 to G61; P13 to T62; P14 to G63; E15 to A64; E16 to F65; V17 to E66; E18 to 167; P19 to E68; G20 to 169;
S21 to N70; G22 to G71; V23 to Q72; R24 to L73; 125 to V74; V26 to F75; V27 to S76; E28 to K77; Y29 to L78; C30 to E79; E31 to N80; P32 to G81; C33 to G82; G34 to F83; F35 to P84; E36 to Y85; A37 to E86; T38 to K87; Y39 to D88;
= MO to L89; E41 to 190; L42 to E91; A43 to A92; S44 to 193; A45 to R94;

to R95; K47 to A96; E48 to S97; Q49 to N98; Y50 to G99; P51 to E100; G52 to T101; 153 to L102; E54 to E103; 155 to K104; E56 to 1105; S57 to T106; R58 to N107; L59 to S108; G60 to R109; G61 to P110; T62 to P111; G63 to C112; A64 to V113; F65 to 1114; E66 to L115 [0157]1 In another preferred embodiment, the isolated polypeptides of the present invention comprising or, alternatively, consisting of, one or more C35 peptide epitopes include the following 5 liners (residues correspond to SEQ ID NO:2 and FIG. 1B):
M1 to P51; S2 to G52; G3 to 153; E4 to E54; P5 to 155; G6 to E56; Q7 to S57;
T8 to R58; S9 to L59; V10 to G60; All to G61; P12 to T62; P13 to G63; P14 to A64; E15 to F65; E16 to E6,6; V17 to 167; E18 to E68; P19 to 169; G20 to N70;
S21 to G71; G22 to Q72; V23 to L73; R24 to V74; 125 to F75; V26 to S76; V27 to K77; E28 to L78; Y29 to E79; C30 to N80; E31 to G81; P32 to G82; C33 to F83; G34 to P84; F35 to Y85; E36 to E86; A37 to K87; T38 to D88; Y39 to L89;
MO to 190; E41 to E91; L42 to A92; A43 to 193; S44 to R94; A45 to R95; V46 to A96; K47 to S97; E48 to N98; Q49 to G99; Y50 to E100; P51 to T101; G52 to L102; 153 to E103; E54 to K104; 155 to 1105; E56 to T106; S57 to N107; R58 to S108; L59 to R109; G60 to P110; G61 to P111; T62 to C112; G63 to V113;
A64 to 1114; F65 to L115 [0158] In another preferred embodiment, the isolated polypeptides of the present invention comprising or, alternatively, consisting of, one or more C35 peptide epitopes include the following 52mers (residues correspond to SEQ ID NO:2 and FIG. 1B):
M1 to G52; S2 to 153; G3 to E54; E4 to 155; P5 to E56; G6 to S57; Q7 to R58;
T8 to L59; S9 to G60; V10 to G61; All to T62; P12 to G63; P13 to A64; P14 to F65; El5 to E66; E16 to 167; V17 to E68; E18 to 169; P19 to N70; G20 to G71;
S21 to Q72; G22 to L73; V23 to V74; R24 to F75; 125 to S76; V26 to K77; V27 to L78; E28 to E79; Y29 to N80; C30 to G81; E31 to G82; P32 to F83; C33 to P84; G34 to Y85; F35 to E86; E36 to K87; A37 to D88; T38 to L89; Y39 to 190;
MO to E91; E41 to A92; L42 to 193; A43 to R94; S44 to R95; A45 to A96; V46 to S97; K47 to N98; E48 to G99; Q49 to E100; Y50 to T101; P51 to L102; G52 to E103; 153 to K104; E54 to 1105; 155 to T106; E56 to N107; S57 to S108; R58 to R109; L59 to P110; G60 to P111; G61 to C112; T62 to V113; G63 to 1114;
A64 to L115 [0159] In another preferred embodiment, the isolated polypeptides of the present invention comprising or, alternatively, consisting of, one or more C35 peptide epitopes include the following 53mers (residues correspond to SEQ ID NO:2 and FIG. 1B):
M1 to 153; S2 to E54; G3 to 155; E4 to E56; P5 to S57; G6 to R58; Q7 to L59;
T8 to G60; S9 to G61; V10 to T62; All to G63; P12 to A64; P13 to F65; P14 to E66; E15 to 167; E16 to E68; V17 to 169; E18 to N70; P19 to G71; G20 to Q72;
S21 to L73; G22 to V74; V23 to F75; R24 to S76; 125 to K77; V26 to L78; V27 to E79; E28 to N80; Y29 to G81; C30 to G82; E31 to F83; P32 to P84; C33 to Y85; G34 to E86; F35 to K87; E36 to D88; A37 to L89; T38 to 190; Y39 to E91;
L40 to A92; X41 to 193; L42 to R94; A43 to R95; S44 to A96; A45 to S97; V46 to N98; K47 to G99; E48 to E100; Q49 to T101; Y50 to L102; P51 to E103; G52 to K104; 153 to 1105; E54 to T106;155 to N107; E56 to S108; S57 to R109; R58 to P110; L59 to P111; G60 to C112; G61 to V113; T62 to 1114; G63 to L115 [0160] In another preferred embodiment, the isolated polypeptides of the present invention comprising or, alternatively, consisting of, one or more C35 peptide epitopes include the following 54mers (residues correspond to SEQ ID NO:2 and FIG. 1B):
M1 to E54; S2 to 155; G3 to E56; E4 to S57; P5 to R58; G6 to L59; Q7 to G60;
T8 to G61; S9 to T62; V10 to G63; All to A64; P12 to F65; P13 to E66; P14 to 167; El5 to E68; El6 to 169; V17 to N70; El8 to G71; P19 to Q72; G20 to L73;
S21 to V74; G22 to F75; V23 to S76; R24 to K77; 125 to L78; V26 to E79; V27 to N80; E28 to G81; Y29 to G82; C30 to F83; E31 to P84; P32 to Y85; C33 to E86; G34 to K87; F35 to D88; E36 to L89; A37 to 190; T38 to E91; Y39 to A92;
L40 to 193; E41 to R94; L42 to R95; A43 to A96; S44 to S97; A45 to N98; V46 to G99; K47 to E100; E48 to T101; Q49 to L102; Y50 to E103; P51 to K104;
G52 to Il05; 153 to T106; E54 to N107;155 to S108; E56 to R109; S57 to P110;
R58 to P111; L59 to C112; G60 to V113; G61 to 1114; T62 to L115 [0161] In another preferred embodiment, the isolated polypeptides of the present invention comprising or, alternatively, consisting of, one or more C35 peptide epitopes include the following 55mers (residues correspond to SEQ ID NO:2 and FIG. 1B):

M1 to 155; S2 to E56; G3 to S57; E4 to R58; P5 to L59; G6 to G60; Q7 to G61;
T8 to T62; S9 to G63; V10 to A64; All to F65; P12 to E66; P13 to 167; P14 to E68; E15 to 169; E16 to N70; V17 to G71; E18 to Q72; P19 to L73; G20 to V74;
S21 to F75; G22 to S76; V23 to K77; R24 to L78; 125 to E79; V26 to N80; V27 to G81; E28 to G82; Y29 to F83; C30 to P84; E31 to Y85; P32 to E86; C33 to K87; G34 to D88; F35 to L89; E36 to 190; A37 to E91; T38 to A92; Y39 to 193;
L40 to R94; E41 to R95; L42 to A96; A43 to S97; S44 to N98; A45 to G99; V46 to E100; K47 to T101; E48 to L102; Q49 to E103; Y50 to K104; P51 to 1105;
G52 to T106;153 to N107; E54 to S108;155 to R109; E56 to P110; S57 to P111;
R58 to C112; L59 to V113; G60 to 1114; G61 to L115 [0162] In another preferred embodiment, the isolated polypeptides of the present invention comprising or, alternatively, consisting of, one or more C35 peptide epitopes include the following 56mers (residues correspond to SEQM NO:2 and FIG. 1B):
M1 to E56; S2 to S57; G3 to R58; E4 to L59; P5 to G60; G6 to G61; Q7 to T62;
T8 to G63; S9 to A64; V10 to F65; All to E66; P12 to 167; P13 to E68; P14 to 169; E15 to N70; E16 to G71; V17 to Q72; E18 to L73; P19 to V74; G20 to F75;
S21 to S76; G22 to K77; V23 to L78; R24 to E79; 125 to N80; V26 to G81; V27 to G82; E28 to F83; Y29 to P84; C30 to Y85; E31 to E86; P32 to K87; C33 to D88; G34 to L89; F35 to 190; E36 to E91; A37 to A92; T38 to 193; Y39 to R94;
L40 to R95; E41 to A96; L42 to S97; A43 to N98; S44 to G99; A45 to E100;
V46 to T101; K47 to L102; E48 to E103; Q49 to K104; Y50 to 1105; P51 to T106; G52 to N107; 153 to S108; E54 to R109; 155 to P110; E56 to P111; S57 to C112; R58 to V113; L59 to 1114; G60 to L115 [0163] In another preferred embodiment, the isolated polypeptides of the present invention comprising or, alternatively, consisting of, one or more C35 peptide epitopes include the following 57mers (residues correspond to SEQ ID NO:2 and FIG. 1B):
M1 to S57; S2 to R58; G3 to L59; E4 to G60; P5 to G61; G6 to T62; Q7 to G63;
T8 to A64; S9 to F65; V10 to E66; All to 167; P12 to E68; P13 to 169; P14 to N70; E15 to G71; E16 to Q72; V17 to L73; E18 to V74; P19 to F75; G20 to S76;
S21 to K77; G22 to L78; V23 to E79; R24 to N80; 125 to G81; V26 to G82; V27 to F83; E28 to P84; Y29 to Y85; C30 to E86; E31 to K87; P32 to D88; C33 to L89; G34 to 190; F35 to E91; E36 to A92; A37 to 193; T38 to R94; Y39 to R95;
L40 to A96; EA-1 to S97; L42 to N98; A43 to G99; S44 to E100; A45 to T101;
V46 to L102; K47 to E103; E48 to K104; Q49 to 1105; Y50 to T106; P51 to N107; G52 to S108; 153 to R109; E54 to P110; 155 to P111; E56 to C112; S57 to V113; R58 to 1114; L59 to L115 [0164] In another preferred embodiment, the isolated polypeptides of the present invention comprising or, alternatively, consisting of, one or more C35 peptide epitopes include the following 58mers (residues correspond to SEQ ID NO:2 and FIG. 1B):
M1 to R58; S2 to L59; G3 to G60; E4 to G61; P5 to T62; G6 to G63; Q7 to A64;
T8 to F65; S9 to E66; V10 to 167; Al 1 to E68; P12 to 169; P13 to N70; P14 to G71; E15 to Q72; El6 to L73; V17 to V74; E18 to F75; P19 to S76; G20 to K77;
S21 to L78; G22 to E79; V23 to N80; R24 to G81; 125 to G82; V26 to F83; V27 to P84; E28 to Y85; Y29 to E86; C30 to K87; E31 to E88; P32 to L89; C33 to 190; G34 to E91; F35 to A92; E36 to 193; A37 to R94; T38 to R95; Y39 to A96;
L40 to S97; El to N98; L42 to G99; A43 to E100; S44 to T101; A45 to L102;
V46 to E103; K47 to K104; E48 to 1105; Q49 to T106; Y50 to N107; P51 to S108; G52 to R109; 153 to P110; E54 to P111; 155 to C112; E56 to V113; S57 to 1114; R58 to L115 [0165] In another preferred embodiment, the isolated polypeptides of the present invention comprising or, alternatively, consisting of, one or more C35 peptide epitopes include the following 59mers (residues correspond to SEQ ID NO:2 and FIG. 1B):
M1 to L59; S2 to G60; G3 to G61; E4 to T62; P5 to G63; G6 to A64; Q7 to F65;
T8 to E66; S9 to 167; V10 to E68; All to 169; P12 to N70; P13 to G71; P14 to Q72; E15 to L73; El6 to V74; V17 to F75; E18 to S76; P19 to K77; G20 to L78;
S21 to E79; G22 to N80; V23 to G81; R24 to G82; 125 to F83; V26 to P84; V27 to Y85; E28 to E86; Y29 to K87; C30 to D88; E31 to L89; P32 to 190; C33 to E91; G34 to A92; F35 to 193; E36 to R94; A37 to R95; T38 to A96; Y39 to S97;
L40 to N98; E41 to G99; L42 to E100; A43 to T101; S44 to L102; A45 to E103;
V46 to K104; K47 to 1105; E48 to T106; Q49 to N107; Y50 to S108; P51 to R109; G52 to P110; 153 to P111; E54 to C112; 155 to V113; E56 to 1114; S57 to [0166] In another preferred embodiment, the isolated polypeptides of the present invention comprising or, alternatively, consisting of, one or more C35 peptide epitopes include the following 60mers (residues correspond to SEQ lD NO:2 and FIG. 1B):
M1 to G60; S2 to G61; G3 to T62; E4 to G63; P5 to A64; G6 to F65; Q7 to E66;
T8 to 167; S9 to E68; V10 to 169; All to N70; P12 to G71; P13 to Q72; P14 to L73; El5 to V74; El6 to F75; V17 to S76; EH to K77; P19 to L78; G20 to E79;
S21 to N80; G22 to G81; V23 to G82; R24 to F83; 125 to P84; V26 to Y85; V27 to E86; E28 to K87; Y29 to D88; C30 to L89; E31 to 190; P32 to E91; C33 to A92; G34 to 193; F35 to R94; E36 to R95; A37 to A96; T38 to S97; Y39 to N98;
L40 to G99; E41 to E100; L42 to T101; A43 to L102; S44 to E103; A45 to K104;
V46 to 1105; K47 to T106; E48 to N107; Q49 to S108; Y50 to R109; P51 to P110; G52 to P111; 153 to C112; E54 to V113; 155 to 1114; E56 to L115 [0167] In another preferred embodiment, the isolated polypeptides ofthe present invention comprising or, alternatively, consisting of, one or more C35 peptide epitopes include the following 61mers (residues correspond to SEQ ID NO:2 and FIG. 1B):
M1 to G61; S2 to T62; G3 to G63; E4 to A64; P5 to F65; G6 to E66; Q7 to 167;
T8 to E68; S9 to 169; V10 to N70; All to G71; P12 to Q72; P13 to L73; P14 to V74; EIS to F75; El6 to S76; V17 to K77; El8 to L78; P19 to E79; G20 to N80;
S21 to G81; G22 to G82; V23 to F83; R24 to P84; 125 to Y85; V26 to E86; V27 to K87; E28 to D88; Y29 to L89; C30 to 190; E31 to E91; P32 to A92; C33 to 193; G34 to R94; F35 to R95; E36 to A96; A37 to S97; T38 to N98; Y39 to G99;
L40 to E100; E41 to T101; L42 to L102; A43 to E103; S44 to K104; A45 to 1105; V46 to T106; K47 toN107; E48 to S108; Q49 to R109; Y50 to P110; P51 to P111; G52 to C112; 153 to V113; E54 to 1114; 155 to L115 [0168] In another preferred embodiment, the isolated polypeptides of the present invention comprising or, alternatively, consisting of, one or more C35 peptide epitopes include the following 62mers (residues correspond to SEQ ID NO:2 and FIG. 1B):
M1 to T62; S2 to G63; G3 to A64; E4 to F65; P5 to E66; G6 to 167; Q7 to E68;
T8 to 169; S9 to N70; V10 to G71; All to Q72; P12 to L73; P13 to V74; P14 to F75; EIS to S76; El6 to K77; V17 to L78; El8 to E79; P19 to N80; G20 to G81;
S21 to G82; G22 to F83; V23 to P84; R24 to Y85;125 to E86; V26 to K87; V27 to D88; E28 to L89; Y29 to 190; C30 to E91; E31 to A92; P32 to 193; C33 to R94; G34 to R95; F35 to A96; E36 to S97; A37 to N98; T38 to G99; Y39 to E100; 1,40 to T101; E41 to L102; 1.42 to E103; A43 to K104; S44 to 1105; A45 to T106; V46 to N107; K47 to S108; E48 to R109; Q49 to P110; Y50 to P111;
P51 to C112; G52 to V113; 153 to 1114; E54 to L115.
[0169] In another preferred embodiment, the isolated polypeptides of the present invention comprising or, alternatively, consisting of, one or more C35 peptide epitopes include the following 63mers (residues correspond to SEQ ID NO:2 and FIG. 1B):
M1 to G63; S2 to A64; G3 to F65; E4 to E66; P5 to 167; G6 to E68; Q7 to 169;
T8 to N70; S9 to G71; V10 to Q72; All to L73; P12 to V74; P13 to F75; P14 to S76; EIS to K77; El6 to L78; V17 to E79; El8 to N80; P19 to G81; G20 to G82;
S21 to F83; G22 to P84; V23 to Y85; R24 to E86; 125 to K87; V26 to D88; V27 to L89; E28 to 190; Y29 to E91; C30 to A92; E31 to 193; P32 to R94; C33 to R95; G34 to A96; F35 to S97; E36 to N98; A37 to G99; T38 to E100; Y39 to T101; 1A0 to L102; E41 to E103; L42 to K104; A43 to 1105; S44 to T106; A45 to N107; V46 to S108; K47 to R109; E48 to P110; Q49 to P111; Y50 to C112;
P51 to V113; G52 to 1114; 153 to L115 [0170] In another preferred embodiment, the isolated polypeptides of the present invention comprising or, alternatively, consisting of, one or more C35 peptide epitopes include the following 64mers (residues correspond to SEQ ID NO:2 and FIG. 1B):
M1 to A64; S2 to F65; G3 to E66; E4 to 167; P5 to E68; G6 to 169; Q7 to N70;
T8 to G71; S9 to Q72; V10 to L73; All to V74; P12 to F75; P13 to S76; P14 to K77; E15 to L78; El6 to E79; V17 to N80; E18 to G81; P19 to G82; G20 to F83;
S21 to P84; G22 to Y85; V23 to E86; R24 to K87; 125 to D88; V26 to L89; V27 to 190; E28 to E91; Y29 to A92; C30 to 193; E31 to R94; P32 to R95; C33 to A96; G34 to S97; F35 to N98; E36 to G99; A37 to E100; T38 to T101; Y39 to L102; IA0 to E103; E41 to K104; L42 to 1105; A43 to T106; S44 to N107; A45 to S108; V46 to R109; K47 to P110; E48 to P111; Q49 to C112; Y50 to V113;
P51 to 1114; G52 to L115;
[0171] In another preferred embodiment, the isolated polypeptides of the present invention comprising or, alternatively, consisting of, one or more C35 peptide epitopes include the following 65mers (residues correspond to SEQ ID NO:2 and FIG. 1B):
M1 to F65; S2 to E66; G3 to 167; E4 to E68; P5 to 169; G6 to N70; Q7 to G71;
T8 to Q72; S9 to L73; V10 to V74; All to F75; P12 to S76; P13 to K77; P14 to L78; E15 to E79; E16 to N80; V17 to G81; E18 to G82; P19 to F83; G20 to P84;
S21 to Y85; G22 to E86; V23 to K87; R24 to D88; 125 to L89; V26 to 190; V27 to E91; E28 to A92; Y29 to 193; C30 to R94; E31 to R95; P32 to A96; C33 to S97; G34 to N98; F35 to G99; E36 to E100; A37 to T101; T38 to L102; Y39 to E103; IA0 to K104; E41 to 1105; L42 to T106; A43 to N107; S44 to S108; A45 to R109; V46 to P110; K47 to P111; E48 to C112; Q49 to V113; Y50 to 1114;
P51 to L115;
[0172] In another preferred embodiment, the isolated polypeptides of the present invention comprising or, alternatively, consisting of, one or more C35 peptide epitopes include the following 66mers (residues correspond to SEQ ID NO:2 and FIG. 1B):
M1 to E66; S2 to 167; G3 to E68; E4 to 169; P5 to N70; G6 to G71; Q7 to Q72;
T8 to L73; S9 to V74; V10 to F75; All to S76; P12 to K77; P13 to L78; P14 to E79; E15 to N80; E16 to G81; V17 to G82; E18 to F83; P19 to P84; G20 to Y85;
S21 to E86; G22 to K87; V23 to D88; R24 to L89; 125 to 190; V26 to E91; V27 to A92; E28 to 193; Y29 to R94; C30 to R95; E31 to A96; P32 to S97; C33 to N98; G34 to G99; F35 to E100; E36 to T101; A37 to L102; T38 to E103; Y39 to K104; IA0 to 1105; E41 to T106; L42 to N107; A43 to S108; S44 to R109;
A45 to P110; V46 to P111; K47 to C112; E48 to V113; Q49 to 1114; Y50 to [0173] In another preferred embodiment, the isolated polypeptides of the present invention comprising or, alternatively, consisting of, one or more C35 peptide epitopes include the following 67mers (residues correspond to SEQ ID NO:2 and FIG. 1B):
M1 to 167; S2 to E68; G3 to 169; E4 to N70; P5 to G71; G6 to Q72; Q7 to L73;
T8 to V74; S9 to F75; V10 to S76; All to K77; P12 to L78; P13 to E79; P14 to N80; E15 to G81; E16 to G82; V17 to F83; E18 to P84; P19 to Y85; G20 to E86;
S21 to K87; G22 to D88; V23 to L89; R24 to 190; 125 to E91; V26 to A92; V27 to 193; E28 to R94; Y29 to R95; C30 to A96; E31 to S97; P32 to N98; C33 to G99; G34 to E100; F35 to T101; E36 to L102; A37 to E103; T38 to K104; Y39 to I105; L40 to T106; E41 to N107; L42 to S108; A43 to R109; S44 to P110; A45 to P111; V46 to C112; K47 to V113; E48 to 1114; Q49 to L115;
[0174] In another preferred embodiment, the isolated polypeptides of the present invention comprising or, alternatively, consisting of, one or more C35 peptide epitopes include the following 68mers (residues correspond to SEQ ID NO:2 and FIG. 1B):
M1 to E68; S2 to 169; G3 to N70; E4 to G71; P5 to Q72; G6 to L73; Q7 to V74;
T8 to F75; S9 to S76; V10 to K77; All to L78; P12 to E79; P13 to N80; P14 to G81; EIS to G82; E16 to F83; V17 to P84; E18 to Y85; P19 to E86; G20 to K87;
S21 to D88; G22 to L89; V23 to 190; R24 to E91; 125 to A92; V26 to 193; V27 to R94; E28 to R95; Y29 to A96; C30 to S97; E31 to N98; P32 to G99; C33 to E100; G34 to T101; F35 to L102; E36 to E103; A37 to K104; T38 to 1105; Y39 to T106; L40 to N107; E41 to S108; L42 to R109; A43 to P110; S44 to P111;
A45 to C112; V46 to V113; K47 to 1114; E48 to L115;
[0175] In another preferred embodiment, the isolated polypeptides of the present invention comprising or, alternatively, consisting of, one or more C35 peptide epitopes include the following 69mers (residues correspond to SEQ ID NO:2 and FIG. 1B):
M1 to 169; S2 to N70; G3 to G71; E4 to Q72; P5 to L73; G6 to V74; Q7 to F75;
T8 to S76; S9 to K77; V10 to L78; Al 1 to E79; P12 to N80; P13 to G81; P14 to G82; E15 to F83; E16 to P84; V17 to Y85; E18 to E86; P19 to K87; G20 to D88;
S21 to L89; G22 to 190; V23 to E91; R24 to A92; 125 to 193; V26 to R94; V27 to R95; E28 to A96; Y29 to S97; C30 to N98; E31 to G99; P32 to E100; C33 to T101; G34 to L102; F35 to E103; E36 to K104; A37 to 1105; T38 to T106; Y39 to N107; 1,40 to S108; E41 to R109; L42 to P110; A43 to P111; S44 to C112;
A45 to V113; V46 to 1114; 1(47 to L115.
[0176] In another preferred embodiment, the isolated polypeptides of the present invention comprising or, alternatively, consisting of, one or more C35 peptide epitopes include the following 70mers (residues correspond to SEQ ID NO:2 and =
FIG. 1B):
M1 to N70; S2 to G71; G3 to Q72; E4 to L73; P5 to V74; G6 to F75; Q7 to S76;
T8 to K77; S9 to L78; V10 to E79; All to N80; P12 to G81; P13 to G82; P14 to F83; E15 to P84; E16 to Y85; V17 to E86; E18 to K87; P19 to D88; G20 to L89;
S21 to 190; G22 to E91; V23 to A92; R24 to 193; 125 to R94; V26 to R95; V27 to A96; E28 to S97; Y29 to N98; C30 to G99; E31 to E100; P32 to T101; C33 to L102; G34 to E103; F35 to 1(104; E36 to 1105; A37 to T106; T38 to N107;
Y39 to S108; L40 to R109; E41 to P110; L42 to P111; A43 to C112; S44 to V113; A45 to 1114; V46 to L115.
[0177] In another preferred embodiment, the isolated polypeptides of the present invention comprising or, alternatively, consisting of, one or more C35 peptide epitopes include the following 7 lmers (residues correspond to SEQ ED NO:2 and FIG. 1B):

M1 to G71; S2 to Q72; G3 to L73; E4 to V74; P5 to F75; G6 to S76; Q7 to K77;
T8 to L78; S9 to E79; V10 to N80; All to G81; P12 to G82; P13 to F83; P14 to P84; E15 to Y85; El6 to E86; V17 to K87; E18 to D88; P19 to L89; G20 to 190;
S21 to E91; G22 to A92; V23 to 193; R24 to R94; 125 to R95; V26 to A96; V27 to S97; E28 to N98; Y29 to G99; C30 to E100; E31 to T101; P32 to L102; C33 to E103; G34 to K104; F35 to 1105; E36 to T106; A37 to N107; T38 to S108;
Y39 to R109; LAO to P110; E41 to P111; L42 to C112; A43 to V113; S44 to 1114; A45 to L115.
[0178] In another preferred embodiment, the isolated polypeptides of the present invention comprising or, alternatively, consisting of, one or more C35 peptide epitopes include the following 72mers (residues correspond to SEQ ID NO:2 and FIG. 1B):
M1 to Q72; S2 to L73; G3 to V74; E4 to F75; P5 to S76; G6 to K77; Q7 to L78;
T8 to E79; S9 to N80; V10 to G81; All to G82; P12 to F83; P13 to P84; P14 to Y85; E15 to E86; E16 to K87; V17 to D88; El8 to L89; P19 to 190; G20 to E91;
S21 to A92; G22 to 193; V23 to R94; R24 to R95; 125 to A96; V26 to S97; V27 to N98; E28 to G99; Y29 to E100; C30 to T101; E31 to L102; P32 to E103; C33 to K104; G34 to 1105; F35 to T106; E36 to N107; A37 to S108; T38 to R109;
Y39 to P110; L40 to P111; E41 to C112; L42 to V113; A43 to I114; S44 toL115.
[0179] In another preferred embodiment, the isolated polypeptides of the present invention comprising or, alternatively, consisting of, one or more C35 peptide epitopes include the following 73mers (residues correspond to SEQ ID NO:2 and FIG. 1B):
M1 to L73; S2 to V74; G3 to F75; E4 to S76; P5 to K77; G6 to L78; Q7 to E79;
T8 to N80; S9 to G81; V10 to G82; All to F83; P12 to P84; P13 to Y85; P14 to E86; EIS to K87; E16 to D88; V17 to L89; E18 to 190; P19 to E91; G20 to A92;
S21 to 193; G22 to R94; V23 to R95; R24 to A96; 125 to S97; V26 to N98; V27 to G99; E28 to E100; Y29 to T101; C30 to L102; E31 to E103; P32 to K104; C33 to 1105; G34 to T106; F35 to N107; E36 to S108; A37 to R109; T38 to P110;
Y39 to P111; LAO to C112; E41 to V113; L42 to 1114; A43 to L115;

[0180] In another preferred embodiment, the isolated polypeptides of the present invention comprising or, alternatively, consisting of, one or more C35 peptide epitopes include the following 74mers (residues correspond to SEQ ID NO:2 and FIG. 1B):
M1 to V74; S2 to F75; G3 to S76; E4 to K77; P5 to L78; G6 to E79; Q7 to N80;
T8 to G81; S9 to G82; V10 to F83; All to P84; P12 to Y85; P13 to E86; P14 to K87; E15 to D88; E16 to L89; V17 to 190; E18 to E91; P19 to A92; G20 to 193;
S21 to R94; G22 to R95; V23 to A96; R24 to S97;125 to N98; V26 to G99; V27 to E100; E28 to T101; Y29 to L102; C30 to E103; E31 to K104; P32 to 1105;
C33 to T106; G34 to N107; F35 to S108; E36 to R109; A37 to P110; T38 to P111; Y39 to C112; L40 to V113; E41 to 1114; L42 to L115.
[0181] In another preferred embodiment, the isolated polypeptides of the present invention comprising or, alternatively, consisting of, one or more C35 peptide epitopes include the following 75mers (residues correspond to SEQ ID NO:2 and FIG. 1B):
M1 to F75; S2 to S76; G3 to 1(77; E4 to L78; P5 to E79; G6 to N80; Q7 to G81;
T8 to G82; S9 to F83; V10 to P84; All to Y85; P12 to E86; P13 to 1(87; P14 to D88; E15 to L89; E16 to 190; V17 to E91; E18 to A92; P19 to 193; G20 to R94;
S21 to R95; G22 to A96; V23 to S97; R24 to N98;125 to G99; V26 to E100; V27 to T101; E28 to L102; Y29 to E103; C30 to K104; E31 to 1105; P32 to T106;
C33 to N107; G34 to S108; F35 to R109; E36 to P110; A37 to P111; T38 to C112; Y39 to V113; MO to 1114; E41 to L115;
[0182] In another preferred embodiment, the isolated polyp eptides of the present invention comprising or, alternatively, consisting of, one or more C35 peptide epitopes include the following 76mers (residues correspond to SEQ ID NO:2 and FIG. 1B):
M1 to S76; S2 to 1(77; G3 to L78; E4 to E79; P5 to N80; G6 to G81; Q7 to G82;
T8 to F83; S9 to P84; V10 to Y85; All to E86; P12 to 1(87; P13 to D88; P14 to L89; E15 to 190; E16 to E91; V17 to A92; E18 to 193; P19 to R94; G20 to R95;
S21 to A96; G22 to S97; V23 to N98; R24 to G99; 125 to E100; V26 to T101;

V27 to L102; E28 to E103; Y29 to K104; C30 to 1105; E31 to T106; P32 to N107; C33 to S108; G34 to R109; F35 to P110; E36 to P111; A37 to C112; T38 to V113; Y39 to 1114; L40 to L115;
[0183] In another preferred embodiment, the isolated polypeptides of the present invention comprising or, alternatively, consisting of, one or more C35 peptide epitopes include the following 77mers (residues correspond to SEQ ID NO:2 and FIG. 1B):
M1 to K77; S2 to L78; G3 to E79; E4 to N80; P5 to G81; G6 to G82; Q7 to F83;
T8 to P84; S9 to Y85; V10 to E86; All to K87; P12 to D88; P13 to L89; P14 to 190; E15 to E91; E16 to A92; V17 to 193; E18 to R94; P19 to R95; G20 to A96;
S21 to S97; G22 to N98; V23 to G99; R24 to E100; 125 to T101; V26 to L102;
V27 to E103; E28 to K104; Y29;1105; C30 to T106; E31 to N107; P32 to S108;
C33 to R109; G34 to P110; F35 to P111; E36 to C112; A37 to V113; T38 to 1114; Y39 to L115;
[0184] In another preferred embodiment, the isolated polypeptides of the present invention comprising or, alternatively, consisting of, one or more C35 peptide epitopes include the following 78mers (residues correspond to SEQ ID NO:2 and FIG. 1B):
M1 to L78; S2 to E79; G3 to N80; E4 to G81; P5 to G82; G6 to F83; Q7 to P84;
T8 to Y85; S9 to E86; V10 to K87; All to D.88; P12 to L89; P13 to 190; P14 to E91; E15 to A92; E16 to 193; V17 to R94; E18 to R95; P19 to A96; G20 to S97;
S21 to N98; G22 to G99; V23 to E100; R24 to T101; 125 to L102; V26 to E103;
V27 to K104; E28 to 1105; Y29 to T106; C30 to N107; E31 to S108; P32 to R109; C33 to P110; G34 to P111; F35 to C112; E36 to V113; A37 to 1114; T38 to L115.
[0185] In another preferred embodiment, the isolated polypeptides of the present invention comprising or, alternatively, consisting of, one or more C35 peptide epitopes include the following 79mers (residues correspond to SEQ ID NO:2 and FIG. 1B):

M1 to E79; S2 to N80; G3 to G81; E4 to G82; P5 to F83; G6 to P84; Q7 to Y85;
T8 to E86; S9 to K87; V10 to D88; All to L89; P12 to 190; P13 to E91; P14 to A92; EIS to 193; E16 to R94; V17 to R95; E18 to A96; P19 to S97; G20 to N98;
S21 to G99; G22 to E100; V23 to T101; R24 to L102;125 to E103; V26 to K104;
V27 to 1105; E28 to T106; Y29 to N107; C30 to S108; E31 to R109; P32 to P110; C33 to P111; G34 to C112; F35 to V113; E36 to 1114; A37 to L115.
[0186] In another preferred embodiment, the isolated polypeptides of the present invention comprising or, alternatively, consisting of, one or more C35 peptide epitopes include the following 80mers (residues correspond to SEQ ID NO:2 and FIG. 1B):
M1 to N80; S2 to G81; G3 to G82; E4 to F83; P5 to P84; G6 to Y85; Q7 to E86;
T8 to K87; S9 to D88; V10 to L89; All to 190; P12 to E91; P13 to A92; P14 to 193; E15 to R94; E16 to R95; V17 to A96; E18 to S97; P19 to N98; G20 to G99;
S21 to E100; G22 to T101; V23 to L102; R24 to E103; 125 to K104; V26 to 1105;
V27 to T106; E28 to N107; Y29 to S108; C30 to R109; E31 to P110; P32 to P111; C33 to C112; G34 to V113; F35 to 1114; E36 to L115.
[0187] In another preferred embodiment, the isolated polypeptides of the present invention comprising or, alternatively, consisting of, one or more C35 peptide = epitopes include the following 8 liners (residues correspond to SEQ ID
NO:2 and FIG. 1B):
M1 to G81; S2 to G82; G3 to F83; E4 to P84; P5 to Y85; G6 to E86; Q7 to K87;
T8 to D88; S9 to L89; V10 to 190; All to E91; P12 to A92; P13 to 193; P14 to R94; E15 to R95; E16 to A96; V17 to S97; E18 to N98; P19 to G99; G20 to = E100; S21 to T101; G22 to L102; V23 to E103; R24 to K104; 125 to 1105;

to T106; V27 to N107; E28 to S108; Y29 to R109; C30 to P110; E31 to P111;
P32 to C112; C33 to V113; G34 to 1114; F35 to L115.
[0188] In another preferred embodiment, the isolated polyp eptides of the present invention comprising or, alternatively, consisting of, one or more C35 peptide epitopes include the following 82mers (residues correspond to SEQ ID NO:2 and FIG. 1B):

M1 to G82; S2 to F83; G3 to P84; E4 to Y85; P5 to E86; G6 to K87; Q7 to D88;
T8 to L89; S9 to 190; V10 to E91; All to A92; P12 to 193; P13 to R94; P14 to R95; E15 to A96; E16 to S97; V17 to N98; El8 to G99; P19 to E100; G20 to T101; S21 to L102; G22 to E103; V23 to K104; R24 to 1105; 125 to T106; V26 to N107; V27 to S108; E28 to R109; Y29 to P110; C30 to P111; E31 to C112;
P32 to V113; C33 to 1114; G34 to L115.
[0189] In another preferred embodiment, the isolated polypeptides of the present invention comprising or, alternatively, consisting of, one or more C35 peptide epitopes include the following 83mers (residues correspond to SEQ ID NO:2 and FIG. 1B):
M1 to F83; S2 to P84; G3 to Y85; E4 to E86; P5 to 1(87; G6 to D88; Q7 to L89;
T8 to 190; S9 to E91; V10 to A92; All to 193; P12 to R94; P13 to R95; P14 to A96; E15 to S97; E16 to N98; V17 to G99; E18 to E100; P19 to T101; G20 to L102; S21 to E103; G22 to K104; V23 to 1105; R24 to T106; 125 to N107; V26 to S108; V27 to R109; E28 to P110; Y29 to P111; C30 to C112; E31 to V113;
P32 to 1114; C33 to L115.
[0190] In another preferred embodiment, the isolated polypeptides of the present invention comprising or, alternatively, consisting of, one or more C35 peptide epitopes include the following 84mers (residues correspond to SEQ ID NO:2 and FIG. 1B):
M1 to P84; S2 to Y85; G3 to E86; E4 to 1(87; P5 to D88; G6 to L89; Q7 to 190;
T8 to E91; S9 to A92; V10 to I93; All to R94; P12 to R95; P13 to A96; P14 to S97; El5 to N98; E16 to G99; V17 to E100; E18 to T101; P19 to L102; G20 to E103; S21 to K104; G22 to 1105; V23 to T106; R24 to N107; 125 to S108; V26 to R109; V27 to P110; E28 to P111; Y29 to C112; C30 to V113; E31 to 1114;
P32 to L115.
[0191] In another preferred embodiment, the isolated polypeptides of the present invention comprising or, alternatively, consisting of, one or more C35 peptide epitopes include the following 85mers (residues correspond to SEQ ID NO:2 and FIG. 1B):

M1 to Y85; S2 to E86; G3 to K87; E4 to D88; P5 to L89; G6 to 190; Q7 to E91;
T8 to A92; S9 to 193; V10 to R94; Al 1 to R95; P12 to A96; P13 to S97; P14 to N98; E15 to G99; E16 to E100; V17 to T101; E18 to L102; P19 to E103; G20 to K104; S21 to 1105; G22 to T106; V23 to N107; R24 to S108; 125 to R109; V26 to P110; V27 to P111; E28 to C112; Y29 to V113; C30 to 1114; E31 to L115.
[0192] In another preferred embodiment, the isolated polypeptides of the present invention comprising or, alternatively, consisting of, one or more C35 peptide epitopes include the following 86mers (residues correspond to SEQ ID NO:2 and FIG. 1B):
M1 to E86; S2 to K87; G3 to D88; E4 to L89; P5 to 190; G6 to E91; Q7 to A92;
T8 to 193; S9 to R94; V10 to R95; Al 1 to A96; P12 to S97; P13 to N98; P14 to G99; E15 to E100; E16 to T101; V17 to L102; E18 to E103; P19 to K104; G20 to 1105; S21 to T106; G22 to N107; V23 to S108; R24 to R109;125 to P110; V26 to P111; V27 to C112; E28 to V113; Y29 to 1114; C30 to L115.
[0193] In another preferred embodiment, the isolated polypeptides of the present invention comprising or, alternatively, consisting of, one or more C35 peptide epitopes include the following 87mers (residues correspond to SEQ ID NO:2 and FIG. 1B):
M1 to K87; S2 to D88; G3 to L89; E4 to 190; P5 to E91; G6 to A92; Q7 to 193;
T8 to R94; S9 to R95; V10 to A96; All to S97; P12 to N98; P13 to G99; P14 to E100; E15 to T101; E16 to L102; V17 to E103; E18 to K104; P19 to 1105; G20 to T106; S21 to N107; G22 to S108; V23 to R109; R24 to P110; 125 to P111;
V26 to C112; V27 to V113; E28 to 1114; Y29 to L115.
[0194] In another preferred embodiment, the isolated polypeptides of the present invention comprising or, alternatively, consisting of, one or more C35 peptide epitopes include the following 88mers (residues correspond to SEQ ID NO:2 and FIG. 1B):
M1 to D88; S2 to L89; G3 to 190; E4 to E91; P5 to A92; G6 to 193; Q7 to R94;
T8 to R95; S9 to A96; V10 to S97; All to N98; P12 to G99; P13 to E100; P14 to T101; El5 to L102; E16 to E103; V17 to K104; E18 to 1105; P19 to T106;

G20 to N107; S21 to S108; G22 to R109; V23 to P110; R24 to P111; 125 to C112; V26 to V113; V27 to 1114; E28 to L115.
[0195] In another preferred embodiment, the isolated polypeptides of the present invention comprising or, alternatively, consisting of, one or more C35 peptide epitopes include the following 89mers (residues correspond to SEQ ID NO:2 and FIG. 1B):
M1 to L89; S2 to 190; G3 to E91; E4 to A92; P5 to 193; G6 to R94; Q7 to R95;
T8 to A96; S9 to S97; V10 to N98; All to G99; P12 to E100; P13 to T101; P14 to L102; E15 to E103; E16 to K104; V17 to 1105; E18 to T106; P19 to N107;
G20 to S108; S21 to R109; G22 to P110; V23 to P111; R24 to C112; 125 to V113; V26 to 1114; V27 to L115.
[0196] In another preferred embodiment, the isolated polypeptides of the present invention comprising or, alternatively, consisting of, one or more C35 peptide epitopes include the following 90mers (residues correspond to SEQ ID NO:2 and FIG. 1B):
M1 to 190; S2 to E91; G3 to A92; E4 to 193; P5 to R94; G6 to R95; Q7 to A96;
T8 to S97; S9 to N98; V10 to G99; All to E100; P12 to T101; P13 to L102;
P14 to E103; EIS to K104; E16 to I105; V17 to T106; E18 to N107; P19 to S108;
G20 to R109; S21 to P110; G22 to P111; V23 to C112; R24 to V113; 125 to 1114; V26 to L115.
[0197] In another preferred embodiment, the isolated polypeptides of the present invention comprising or, alternatively, consisting of, one or more C35 peptide epitopes include the following 91mers (residues correspond to SEQ ID NO:2 and FIG. 1B):
M1 to E91; S2 to A92; G3 to 193; E4 to R94; P5 to R95; G6 to A96; Q7 to S97;
T8 to N98; S9 to G99; V10 to E100; All to T101; P12 to L102; P13 to E103;
P14 to K104; E15 to 1105; E16 to T106; V17 to N107; E18 to S108; P19 to R109; G20 to P110; S21 to P111; G22 to C112; V23 to V113; R24 to 1114; 125 to L115.

[0198] In another preferred embodiment, the isolated polypeptides of the present invention comprising or, alternatively, consisting of, one or more C35 peptide epitopes include the following 92mers (residues correspond to SEQ ID NO:2 and FIG. 1B):
M1 to A92; S2 to 193; G3 to R94; E4 to R95; P5 to A96; G6 to S97; Q7 to N98;
T8 to G99; S9 to E100; V10 to T101; All to L102; P12 to E103; P13 to K104;
P14 to I105; EIS to T106; E16 to N107; V17 to S108; E18 to R109; P19 to P110;
G20 to P111; S21 to C112; G22 to V113; V23 to 1114; R24 to L115.
[0199] In another preferred embodiment, the isolated polypeptides of the present invention comprising or, alternatively, consisting of, one or more C35 peptide epitopes include the following 93mers (residues correspond to SEQ ID NO:2 and FIG. 1B):
M1 to 193; S2 to R94; G3 to R95; E4 to A96; P5 to S97; G6 to N98; Q7 to G99;
T8 to E100; S9 to T101; V10 to L102; All to E103; P12 to K104; P13 to I105;
P14 to T106; E15 to N107; E16 to 5108; V17 to R109; E18 to P110; P19 to P111; G20 to C112; S21 to V113; G22 to 1114; V23 to L115.
[0200] In another preferred embodiment, the isolated polypeptides of the present invention comprising or, alternatively, consisting of, one or more C35 peptide epitopes include the following 94mers (residues correspond to SEQ ID NO:2 and FIG. 1B):
M1 to R94; S2 to R95; G3 to A96; E4 to S97; P5 to N98; G6 to G99; Q7 to E100; T8 to T101; S9 to L102; V10 to E103; All to K104; P12 to 1105; P13 to T106; P14 to N107; EIS to S108; El6 to R109; V17 to P110; El8 to P111; P19 to C112; G20 to V113; S21 to 1114; G22 to L115.
[0201] In another preferred embodiment, the isolated polypeptides of the present invention comprising or, alternatively, consisting of, one or more C35 peptide epitopes include the following 95mers (residues correspond to SEQ ID NO:2 and FIG. 1B): M1 to R95; S2 to A96; G3 to S97; E4 to N98; P5 to G99; G6 to E100;
Q7 to T101; T8 to L102; S9 to E103; V10 to K104; All to 1105; P12 to T106;

P13 to N107; P14 to S108; E15 to R109; E16 to P110; V17 to P111; E18 to C112; P19 to V113; G20 to I114; S21 to L115.
[0202] In another preferred embodiment, the isolated polyp eptides of the present invention comprising or, alternatively, consisting of, one or more C35 peptide epitopes include the following 96mers (residues correspond to SEQ ID NO:2 and FIG. 1B):
M1 to A96; S2 to S97; G3 to N98; E4 to G99; P5 to E100; G6 to T101; Q7 to L102; T8 to E103; S9 to K104; V10 to 1105; All to T106; P12 to N107; P13 to S108; P14 to R109; EIS to P110; E16 to P111; V17 to C112; E18 to V113; P19 to 1114; G20 to L115.
[0203] In another preferred embodiment, the isolated polypeptides of the present invention comprising or, alternatively, consisting of, one or more C35 peptide epitopes include the following 97mers (residues correspond to SEQ ID NO:2 and FIG. 1B):
M1 to S97; S2 to N98; G3 to G99; E4 to E100; P5 to T101; G6 to L102; Q7 to E103; T8 to K104; S9 to 1105; V10 to T106; All to N107; P12 to S108; P13 to R109; P14 to P110; EIS to P111; E16 to C112; V17 to V113; El8 to 1114; P19 to L115.
[0204] In another preferred embodiment, the isolated polypeptides of the present invention comprising or, alternatively, consisting of, one or more C35 peptide epitopes include the following 98mers (residues correspond to SEQ ID NO:2 and FIG. 1B):
M1 to N98; S2 to G99; G3 to E100; E4 to T101; P5 to L102; G6 to E103; Q7 to K104; T8 to 1105; S9 to T106; V10 to N107; All to S108; P12 to R109; P13 to P110; P14 to P111; E15 to C112; E16 to V113; V17 to 1114; E18 to L115.
[0205] In another preferred embodiment, the isolated polypeptides of the present invention comprising or, alternatively, consisting of, one or more C35 peptide epitopes include the following 99mers (residues correspond to SEQ JD NO:2 and FIG. 1B):

M1 to G99; S2 to E100; G3 to T101; E4 to L102; P5 to E103; G6 to K104; Q7 to 1105; T8 to T106; S9 to N107; V10 to S108; Al 1 to R109; P12 to P110; P13 to P111; P14 to C112; E15 to V113; El6 to 1114; V17 to L115.
[0206] In another preferred embodiment, the isolated polypeptides of the present invention comprising or, alternatively, consisting of, one or more C35 peptide epitopes include the following 100mers (residues correspond to SEQ ID NO:2 and FIG. 1B):
M1 to E100; S2 to T101; G3 to L102; E4 to E103; P5 to K104; G6 to 1105; Q7 to T106; T8 to N107; S9 to S108; V10 to R109; All to P110; P12 to P111; P13 to C112; P14 to V113; E15 to 1114; E16 to L115.
[0207] In another preferred embodiment, the isolated polypeptides of the present invention comprising or, alternatively, consisting of, one or more C35 peptide epitopes include the following 101mers (residues correspond to SEQ ID NO:2 and FIG. 1B):
M1 to T101; S2 to L102; G3 to E103; E4 to K104; P5 to 1105; G6 to T106; Q7 toN107; T8 to S108; S9 to R109; V10 to P110; All to P111; P12 to C112; P13 to V113; P14 to 1114; E15 to L115.
[0208] In another preferred embodiment, the isolated polypeptides of the present invention comprising or, alternatively, consisting of, one or more C35 peptide epitopes include the following 102mers (residues correspond to SEQ ID NO:2 and FIG. 1B):
M1 to L102; S2 to E103; G3 to K104; E4 to 1105; P5 to T106; G6 to N107; Q7 to S108; T8 to R109; S9 to P110; V10 toP111; All to C112; P12 to V113; P13 to 1114; P14 to L115.
[0209] In another preferred embodiment, the isolated polypeptides of the present invention comprising or, alternatively, consisting of, one or more C35 peptide epitopes include the following 103mers (residues correspond to SEQ ID NO:2 and FIG. 1B):

M1 to E 103; S2 to K104; G3 to 1105; E4 to T106; P5 to N107; G6 to S108; Q7 to R109; T8 to P110; S9 to P111; V10 to C112; All to V113; P12 to 1114; P13 to L115.
[0210] In another preferred embodiment, the isolated polypeptides of the present invention comprising or, alternatively, consisting of, one or more C35 peptide epitopes include the following 104mers (residues correspond to SEQ ID NO:2 and FIG. 1B):
M1 to K104; S2 to 1105; G3 to T106; E4 to N107; P5 to S108; G6 to R109; Q7 to P110; T8 to P111; S9 to C112; V10 to V113; All to 1114; P12 to L115.
[0211] In another preferred embodiment, the isolated polypeptides of the present invention comprising or, alternatively, consisting of, one or more C35 peptide epitopes include the following 105mers (residues correspond to SEQ ID NO:2 and FIG. 1B):
M1 to 1105; S2 to T106; G3 to N107; E4 to S108; P5 to R109; G6 to P110; Q7 to P111; T8 to C112; S9 to V113; V10 to 1114; All to L115.
[0212] In another preferred embodiment, the isolated polypeptides of the present invention comprising or, alternatively, consisting of, one or more C35 peptide epitopes include the following 106mers (residues correspond to SEQ ID NO:2 and FIG. 1B):
M1 to T106; S2 to N107; G3 to S108; E4 to R109; P5 to P110; G6 to P111; Q7 to C112; T8 to V113; S9 to 1114; V10 to L115.
[0213] In another preferred embodiment, the isolated polypeptides of the present invention comprising or, alternatively, consisting of, one or more C35 peptide epitopes include the following 107mers (residues correspond to SEQ ID NO:2 and FIG. 1B):
M1 to N107; S2 to S108; G3 to R109; E4 to P110; P5 to P111; G6 to C112; Q7 to V113; T8 to 1114; S9 to L115.
[0214] In another preferred embodiment, the isolated polypeptides of the present invention comprising or, alternatively, consisting of, one or more C35 peptide epitopes include the following 108mers (residues correspond to SEQ ID NO:2 and FIG. 1B):
M1 to S108; S2 to R109; G3 to P110; E4 to P111; P5 to C112; G6 to V113; Q7 to 1114; T8 to L115.
[02151 In another preferred embodiment, the isolated polypeptides of the present invention comprising or, alternatively, consisting of, one or more C35 peptide epitopes include the following 109mers (residues correspond to SEQ ID NO:2 and FIG. 1B):
M1 to R109; S2 to P110; G3 to P111; E4 to C112; P5 to V113; G6 to 1114; Q7 to L115.
[0216] In another preferred embodiment, the isolated polyp eptides of the present invention comprising or, alternatively, consisting of, one or more C35 peptide epitopes include the following 110mers (residues correspond to SEQ ID NO:2 and FIG. 1B):
M1 to P110; S2 to P111; G3 to C112; E4 to V113; P5 to 1114; G6 to L115.
[02171 In another preferred embodiment, the isolated polypeptides of the present invention comprising or, alternatively, consisting of, one or more C35 peptide epitopes include the following 111mers (residues correspond to SEQ ID NO:2 and FIG. 1B):
M1 to P111; S2 to C112; G3 to V113; E4 to 1114; P5 to L115.
[0218] In another preferred embodiment, the isolated polyp eptides of the present invention comprising or, alternatively, consisting of, one or more C35 peptide epitopes include the following 112mers (residues correspond to SEQ ID NO:2 and FIG. 1B):
M1 to C112; S2 to V113; G3 to 1114; E4 to L115.
[02191 In another preferred embodiment, the isolated polypeptides of the present invention comprising or, alternatively, consisting of, one or more C35 peptide epitopes include the following 113mers (residues correspond to SEQ ID NO:2 and FIG. 1B):
M1 to V113; S2 to 1114; G3 to L115.

[0220] In another preferred embodiment, the isolated polypeptides of the present invention comprising or, alternatively, consisting of, one or more C35 peptide epitopes include the following 114mers (residues correspond to SEQ ID NO:2 and FIG. 1B):
M1 to 1114; S2 to L115.
Stimulation of CTL and HTL responses 102211 Much more about the mechanism by which T cells recognize antigens has been elucidated during the past ten years. In accordance with this understanding of the immune system, the present inventors have developed efficacious peptide epitope compositions that induce a therapeutic or prophylactic immune response to certain tumor associated antigens, when administered via various art-accepted modalities. Moreover, by use of the peptide epitopes of the invention, or by use of combinations of peptide epitopes in accordance with the principles disclosed herein, responses can be achieved in significant percentages of a non-genetically biased worldwide population. For an understanding of the value and efficacy of the claimed compositions, a brief review of immunology-related technology is.
provided.
[0222] A complex of an HLA molecule and a peptidic antigen acts as the ligand recognized by HLA-restricted T cells (Buus, S. et al., Cell 47:1071, 1986;
Babbitt, B. P. et al., Nature 317:359, 1985; Townsend, A. and Bodmer, H., Annu.
Rev. Immunol. 7:601, 1989; Germain, R. N., Annu. Rev. Imnzunol . 11:403, 1993).
Through the study of single amino acid substituted antigen analogs and the sequencing of endogenously bound, naturally processed peptides, critical residues that correspond to motifs required for specific binding to HLA antigen molecules have been identified.
[0223]
Furthermore, x-ray crystallographic analyses of HLA-peptide complexes have revealed pockets within the peptide binding cleft of HLA molecules which accommodate, often on an allele-specific basis, residues borne by peptide ligands;

these residues in turn determine the HLA binding capacity of the peptides in which they are present. (See, e.g., Madden, D.R. Annu. Rev. Immunol. 13:587, 1995; Smith, et aL , Immunity 4:203, 1996; Fremont et aL, Immunity 8:305, 1998;
Stem et a/ ., Structure 2:245, 1994; Jones, E.Y. Gun-. Opin. ImmunoL 9:75, 1997;
Brown, J. H. et al., Nature 364:33, 1993; Guo, H. C. et al.,Proc. Natl. Acad.
Sci.
USA 90:8053, 1993; Guo, H. C. et al., Nature 360:364, 1992; Silver, M. L. et al., Nature 360:367, 1992; Matsumura, M. et al., Science 257:927, 1992; Madden et al., Cell 70:1035, 1992; Fremont, D. H. et al., Science 257:919, 1992; Saper, M.
A. , Bjorla-nan, P. J. and Wiley, D. C., J. Mol. Biol. 219:277,1991.) Accordingly, the definition of class I and class II allele-specific HLA binding motifs, or class I
or class II supermotifs allows identification of regions within a protein that have the predicted ability to bind particular HLA antigen(s).
[0224] Moreover, the correlation of binding affinity with immunogenicity, which is disclosed herein, is an important factor to be considered when evaluating candidate peptides. Thus, by a combination of motif searches of antigenic sequences, and by HLA-peptide binding assays, epitope-based vaccines have been identified. As appreciated by one in the art, after determining their binding affinity, additional work can be performed to select, amongst these vaccine peptides, e.g., epitopes can be selected having optional characteristics in terms of population coverage, antigenicity, and immunogenicity, etc.
[0225] Various strategies can be utilized to evaluate immunogenicity, including:
1) Evaluation ofprimary T cell cultures from normal individuals (see, e.g., Wentworth, P. A. et al., Mol. .1thmunoL 32:603, 1995; Celis, E. et al., Proc.
Natl. Acad. Sci. USA 91:2105, 1994; Tsai, V. et al., J. ImmunoL 158:1796, 1997;
Kawashima, I. et al., Human ImmunoL 59:1, 1998). This procedure involves the stimulation of peripheral blood lymphocytes (PBL) from normal subjects with a test peptide in the presence of antigen presenting cells in vitro over a period of several weeks. T cells specific for the peptide become activated during this time and are detected using, e.g., a 51Cr-release assay involving peptide sensitized target cells, and/or target cells that generate antigen endogenously.

2) Immunization of HLA transgenic mice (see, e.g., Wentworth, P.
A. et aL, J. ImmunoL 26:97, 1996; Wentworth, P. A. et al., Int. ImmunoL 8:651, 1996; Alexander, J. et aL , J. ImmunoL 159:4753, 1997); in this method, peptides in incomplete Freund's adjuvant are administered subcutaneously to HLA
transgenic mice. Several weeks following immunization, splenocytes are removed and cultured in vitro in the presence of test peptide for approximately one week. Peptide-specific T cells are detected using, e.g., a 51Cr-release assay involving peptide sensitized target cells and target cells expressing endogenously generated antigen.
3) Demonstration ofrecall T cell responses from individuals exposed to the disease, such as immune individuals who were effectively treated and recovered from disease, and/or from actively ill patients (see, e.g., Rehermann, B. et al., J. Exp. Med. 181:1047, 1995; Doolan, D. L. et al., Immunity 7:97, 1997;
Bertoni, R. et al., J. ain. Invest. 100:503, 1997; Threlkeld, S. C. et al., J.

ImmunoL 159:1648, 1997; Diepolder, H. M. et al., J. ViroL 71:6011, 1997). In applying this strategy, recall responses are detected by culturing PBL from subjects in vitro for 1-2 weeks in the presence of a test peptide plus antigen presenting cells (APC) to allow activation of "memory" T cells, as compared to "naive" T cells. At the end of the culture period, T cell activity is detected using assays for T cell activity including 51Cr release involving peptide-sensitized targets, T cell proliferation, or lymphokine release.
[02261 The following describes the peptide epitopes and corresponding nucleic acids of the invention in more detail.
Binding Affinity of Peptide Epitopes for HLA Molecules [02271 As indicated herein, the large degree of HLA polymorphism is an important factor to be taken into account with the epitope-based approach to vaccine development. To address this factor, epitope selection encompassing identification of peptides capable of binding at high or intermediate affinity to multiple HLA molecules is preferably utilized, most preferably these epitopes bind at high or intermediate affinity to two or more allele-specific HLA
molecules.
[0228] CTL-inducing peptide epitopes of interest for vaccine compositions preferably include those that have an IC50 or binding affinity value for a class I
HLA molecule(s) of 500 nM or better (i.e., the value is 500 nM). HTL-inducing peptide epitopes preferably include those that have an IC50 or binding affinity value for class 11 HLA molecules of 1000 nM or better, (i.e., the value is 1,000 nM). For example, peptide binding is assessed by testing the capacity of a candidate peptide to bind to a purified HLA molecule in vitro. Peptides exhibiting high or intermediate affinity are then considered for further analysis.
Selected peptides are generally tested on other members of the supertype family.
In preferred embodiments, peptides that exhibit cross-reactive binding are then used in cellular screening analyses or vaccines.
[0229] The relationship between binding affinity for HLA class I molecules and immunogenicity of discrete peptide epitopes on bound antigens has been determined. As disclosed in greater detail herein, higher HLA binding affinity is correlated with greater immunogenicity.
[0230] Greater immunogenicity can be manifested in several different ways.
Immunogenicity corresponds to whether an immune response is elicited at all, and to the vigor of any particular response, as well as to the extent of a population in which a response is elicited. For example, a peptide epitope might elicit an immune response in a diverse array of the population, yet in no instance produce a vigorous response. In accordance with these principles, close to 90% of high binding peptide have been found to elicit a response and thus be "immunogenic,"
as contrasted with about 50% of the peptides that bind with intermediate affinity.
(See, e.g., Schaeffer et al. PNAS 1988) Moreover, not only did peptides with higher binding affinity have an enhanced probability of generating an immune response, the generated response tended to be more vigorous than the response seen with weaker binding peptides. As a result, less peptide is required to elicit a similar biological effect if a high affinity binding peptide is used rather than a lower affinity one. Thus, in preferred embodiments ofthe invention, high affinity binding epitopes are used.
[0231] The correlation between binding affinity and immunogenicity was analyzed by two different experimental approaches (see, e.g., Sette, et al., J.
Immunol. 153:5586-5592, 1994)). In the first approach, the immunogenicity of potential epitopes ranging in HLA binding affinity over a 10,000-fold range was analyzed in HLA-A*0201 transgenic mice. In the second approach, the antigenicity of approximately 100 different hepatitis B virus (HBV)-derived potential epitopes, all carrying A*0201 binding motifs, was assessed by using PBL from acute hepatitis patients. Pursuant to these approaches, it was determined that an affinity threshold value of approximately 500 nM
(preferably 50 nM or less) determines the capacity of a peptide epitope to elicit a CTL
response. These data are true for class I binding affinity measurements for naturally processed peptide epitopes and for synthesized T cell epitopes.
These data also indicate the important role of determinant selection in the shaping of T
cell responses (see, e.g., Schaeffer et al. Proc. Natl. Acad. Sci. USA 86:4649-4653, 1989).
[0232] An affinity threshold associated with immunogenicity in the context of HLA class If (i.e., HLA DR) molecules has also been delineated (see, e.g., Southwood et al. J. Immunology 160:3363-3373,1998. In order to define a biologically significant threshold of HLA class 11 binding affinity, a database of the binding affinities of 32 DR-restricted epitopes for their restricting element (i.e., the HLA molecule that binds the epitope) was compiled. In approximately half of the cases (15 of 32 epitopes), DR restriction was associated with high binding affinities, i.e. binding affinity values of 100 nM or less. In the other half of the cases (16 of 32), DR restriction was associated with intermediate affinity (binding affinity values in the 100-1000 nM range). In only one of 32 cases was DR restriction associated with an IC50 of 1000 nM or greater. Thus, 1000 nM is defined as an affinity threshold associated with immunogenicity in the context of DR molecules.
[0233] Vaccines of the present invention may also comprise epitopes that bind to MHC class II DR molecules. A greater degree of heterogeneity in both size and binding frame position of the motif, relative to the N and C termini of the peptide, exists for class II peptide ligands. This increased heterogeneity of HLA
class 11 peptide ligands is due to the structure of the binding groove of the HLA
class II molecule which, unlike its class I counterpart, is less physically constricted at both ends.
[0234] There are numerous additional supermotifs and motifs in addition to the A2 supermotif and the A2.1-allele specific motif. By inclusion of one or more epitopes from other motifs or supermotifs, enhanced population coverage for major global ethnicities can be obtained.
Peptide Analogs [0235] In general, CTL and HTL responses are not directed against all possible epitopes. Rather, they are restricted to a few "immunodominant" determinants (Zinkernagel, et al., Adv. Immunol. 27:5159, 1979; Bennink, et al., J. Exp.
Med.
168:19351939, 1988; Rawle, et al., J. Immunol. 146:3977-3984, 1991). It has been recognized that immunodominance (Benacerraf, et al., Science 175:273-279, 1972) could be explained by either the ability of a given epitope to selectively bind a particular HLA protein (determinant selection theory) (Vitiello, et al., J.
Immunol. 131:1635, 1983); Rosenthal, et al., Nature 267:156-158, 1977), or to be selectively recognized by the existing TCR (T cell receptor) specificities (repertoire theory) (Klein, J., IMMUNOLOGY, THE SCIENCE OF SELFNONSELF
DISCRIMINATION, John Wiley & Sons, New York, pp. 270-310, 1982). It has been demonstrated that additional factors, mostly linked to processing events, can also play a key role in dictating, beyond strict immunogenicity, which of the many potential determinants will be presented as immunodominant (Sercarz, et aL
Annu. Rev. ImmunoL 11:729-766, 1993).
[0236] The concept of dominance and subdominance is relevant to immunotherapy of both infectious diseases and malignancies. For example, in the course of chronic viral disease, recruitment of subdominant epitopes can be important for successful clearance of the infection, especially if dominant CTL
or HTL specificities have been inactivated by functional tolerance, suppression, mutation of viruses and other mechanisms (Franco, et aL, Curr. Opin. ImmunoL
7:524-531, 1995). In the case of cancer and tumor antigens, CTLs recognizing at least some of the highest binding affinity peptides might be functionally inactivated. Lower binding affinity peptides are preferentially recognized at these times, and may therefore be preferred in therapeutic or prophylactic anti-cancer vaccines.
[0237] In particular, it has been noted that a significant number of epitopes derived from known non-viral tumor associated antigens (TAA) bind HLA class I with intermediate affinity (IC50 in the 50-500 nM range) rather than at high affinity (IC50 of less than 50 nM).
[0238] For example, it has been found that 8 of 15 known TAA peptides recognized by tumor infiltrating lymphocytes (TIL) or CTL bound in the 50-500 nM range. (These data are in contrast with estimates that 90% of known viral antigens were bound by HLA class I molecules with IC50 of 50 nM or less, while only approximately 10% bound in the 50-500 nM range (Sette, et al., J ImmunoL, 153:558-5592, 1994). In the cancer setting this phenomenon is probably due to elimination or functional inhibition of the CTL recognizing several of the highest binding peptides, presumably because of T cell tolerization events.
[0239] Without intending to be bound by theory, it is believed that because T
cells to dominant epitopes may have been clonally deleted, and selecting subdominant epitopes may allow existing T cells to be recruited, which will then lead to a therapeutic or prophylactic response. However, the binding of HLA
molecules to subdominant epitopes is often less vigorous than to dominant ones.

[0240] Accordingly, there is a need to be able to modulate the binding affinity of particular immunogenic epitopes for one or more HLA molecules, to thereby modulate the immune response elicited by the peptide, for example to prepare analog peptides which elicit a more vigorous response. This ability to modulate both binding affinity and the resulting immune response in accordance with these principles greatly enhances the usefulness of peptide epitope-based vaccines and therapeutic agents.
[0241] Although peptides with suitable cross-reactivity among all alleles of a superfamily are identified by the screening procedures described above, cross-reactivity is not always as complete as possible, and in certain cases procedures to increase cross-reactivity of peptides can be useful; moreover, such procedures can also be used to modify other properties of the peptides such as binding affinity or peptide stability. Having established the general rules that govern cross-reactivity of peptides for HLA alleles within a given motif or supermotif, modification (i.e., analoging) of the structure of peptides of particular interest in order to achieve broader (or otherwise modified) HLA binding capacity can be performed. More specifically, peptides that exhibit the broadest cross-reactivity patterns, can be produced in accordance with the teachings herein.
[0242] In brief, the analoging strategy utilizes the motifs or supermotifs that correlate with binding to certain HLA molecules. Analog peptides can be created by substituting amino acid residues at primary anchor, secondary anchor, or at primary and secondary anchor positions. Generally, analogs are made for peptides that already bear a motif or supermotif. For a number of the motifs or supermotifs, residues are defined which are deleterious to binding to allele-specific HLA molecules or members of HLA supertypes that bind the respective motif or supermotif. Accordingly, removal of such residues that are detrimental to binding can be performed. For example, in the case of the A3 sup ertype, when all peptides that have such deleterious residues are removed from the population of peptides used in the analysis, the incidence of cross-reactivity increased from 22% to 37% (see, e.g., Sidney, J. et al., Hu. Immunol. 45:79, 1996). Examples of C35 peptide epitope analogs of the present invention are found in Table 4.
In a particularly preferred embodiment, the isolated polyp eptides of the present invention comprise or, alternatively, consist of the following C35 peptide epitope analogs: for the peptide epitope G22 to C30 of SEQ ID NO:2 and FIG. 1B (i.e., GVRIVVEYC), the analog with either alanine or glycine substituted for cysteine at the ninth amino acid residue (i.e., GVRIVVEYA or GVRIVVEYG); for the peptide epitope 125 to C33 of SEQ ID NO:2 and FIG. 1B (i.e., IVVEYCEPC), the analog with either alanine or glycine substituted for the cysteine at the sixth amino acid residue and/or the ninth amino acid residue (i.e., IVVEYAEPC, IVVEYCEPA IVVEYGEPC, IVVEYCEPG, IVVEYAEPA, IVVEYAEPG, IVVEYGEPA, IVVEYGEPG); for the peptide epitope K77 to Y85 of SEQ ID
NO: 2 and FIG. 1B (i.e., KLENGGFPY), the analog with valine substituted for tyrosine at the ninth amino acid residue (i.e., KLENGGFPY); for peptide epitope K104 to C112 of SEQ ID NO:2 and FIG. 1B (i.e., KITNSRPPC), the analogs with alanine, glycine or leucine substituted for cysteine at the ninth amino acid residue (i.e., KITNSRPPL, KITNSRPPA, KITNSRPPQ); for peptide epitope K104 to V113 of SEQ ID NO:2 and FIG. 1B (i.e., KITNSRPPCV), the analogs with alanine, glycine, serine or leucine substituted for cysteine at the ninth amino acid residue (i.e., KITNSRPPLV, KITNSRPPAV, KITNSRPPGV, KITNSRPPSV), for the peptide epitope 1105 to V113 of SEQ ID NO:2 and FIG.
1B (i.e., 1TNSRPPCV), the analogs wherein either leucine or methionine is substituted for threonine at the second amino acid residue and/or alanine, serine or glycine is substituted for cysteine at the eighth amino acid residue (i.e., lLNSRPPCV, IMNSRPPCV, ITNSRPPAV, ITNSRPPGV, ILNSRPPAV, ILNSRPPGV, IMNSRPPAV, IMNSRPPGV, ILNSRPPSV, IMNSRPPSV, ITNSRPPSV), for the peptide epitope N107 to L115 of SEQ ID NO:2 and FIG.
1B (i.e., NSRPPCML), the analog with either alanine or glycine substituted for cysteine at the sixth amino acid residue (i.e., NSRPPAVIL, NSRPPGVIL). The invention is further directed to polypeptides comprising or, alternatively, consisting of one or more C35 epitope analogs. In a preferred embodiment, the invention is directed to polypeptides comprising one or more C35 epitope analogs and, in addition, one or more C35 peptide epitopes. In a particularly preferred embodiment, the invention is directed to a fusion protein comprising at least one C35 peptide epitope analog selected from the group consisting of: for the peptide epitope G22 to C30 of SEQ ID NO:2 and FIG. 1B (i.e., GVRIVVEYC), the analog with either alanine or glycine substituted for cysteine at the ninth amino acid residue (i.e., GVR1VVEYA or GVRIVVEY); for the peptide epitope 125 to C33 of SEQ ID NO:2 and FIG. 1B (i.e., TVVEYCEPC), the analog with either alanine or glycine substituted for the cysteine at the sixth amino acid residue and/or the ninth amino acid residue (i.e., IVVEYAEPC, 1VVEYCEPA, TVVEYGEPC, 1VVEYCEPG, IVVEYAEPA, IVVEYAEPG, IVVEYGEPA, IVVEYGEP_Q); for the peptide epitope K77 to Y85 of SEQ ID NO: 2 and FIG.
1B (i.e., KLENGGFPY), the analog with valine substituted for tyrosine at the ninth amino acid residue (i.e., KLENGGFPN1); for peptide epitope K104 to C112 of SEQ ID NO:2 and FIG. 1B (i.e., KITNSRPPC), the analogs with alanine, glycine or leucine substituted for cysteine at the ninth amino acid residue (i.e., KITNSRPPL, KITNSRPPA, KITNSRPPG); for peptide epitope K104 to V113 of SEQ ID NO:2 and FIG. 1B (i.e., KITNSRPPCV), the analogs with alanine, glycine, serine or leucine substituted for cysteine at the ninth amino acid residue (i.e., KITNSRPPLV, KITNSRPPAV, KITNSRPPGV, KITNSRPPSV); for the peptide epitope 1105 to V113 of SEQ ID NO:2 and FIG. 1B (i.e., ITNSRPPCV), the analogs wherein either leucine or methionine is substituted for threonine at the second amino acid residue and/or alanine, serine or glycine is substituted for cysteine at the eighth amino acid residue (i.e., ILNSRPPCV, IMNSRPPCV, ITNSRPPAV, ITNSRPPGV, ILNSRPPAV, TLNSRPPGV, ININSRPPAV, IMNSRPPGV, ELNSRPPSV, IMNSRPPSV, ITNSRPPSV), for the peptide epitope N107 to L115 of SEQ ID NO:2 and FIG. 1B (i.e., NSRPPCVIL), the analog with either alanine or glycine substituted for cysteine at the sixth amino acid residue (i.e., NSRPPAVIL, NSRPPGVIL), and at least one C35 peptide epitope selected from the group consisting of: amino acids E4 to P12 of SEQ ID

NO:2, S9 to V17 of SEQ ID NO: 2, S21 to Y29 of SEQ ID NO:2, G22 to C30 of SEQ ID NO: 2,125 to C33 of SEQ ID NO:2, T38 to V46 of SEQ ID NO:2, G61 to 169 of SEQ JD NO:2, T62 to N70 of SEQ ID NO:2, G63 to G71 of SEQ ID
NO:2, F65 to L73 of SEQ ID NO: 2,167 to F75 of SEQ ID NO:2, K77 to Y85 of SEQ ID NO:2, Q72 to E86 of SEQ ID NO:2, G81 to L89 of SEQ ID NO:2, G99 to V113 of SEQ ID NO:2, E100 to V113 of SEQ ID NO:2, K104 to C112 of SEQ
ID NO:2, K104 to V113 of SEQ ID NO: 2, 1105 to V113 of SEQ ID NO:2, and N107 to L115 of SEQ ID NO:2.
[0243] Thus, one strategy to improve the cross-reactivity of peptides within a given supermotif is simply to delete one or more of the deleterious residues present within a peptide and substitute a small "neutral" residue such as Ala (that may not influence T cell recognition of the peptide). An enhanced likelihood of cross-reactivity is expected if, together with elimination of detrimental residues within a peptide, "preferred" residues associated with high affinity binding to an allele-specific HLA molecule or to multiple HLA molecules within a superfamily are inserted.
[0244] To ensure that an analog peptide, when used as a vaccine, actually elicits a CTL response to the native epitope in vivo (or, in the case of class II
epitopes, elicits helper T cells that cross-react with the wild type peptides), the analog peptide may be used to induce T cells in vitro from individuals of the appropriate HLA allele. Thereafter, the immunized cells' capacity to lyse wild type peptide sensitized target cells is evaluated. Alternatively, evaluation of the cells' activity can be evaluated by monitoring [UN release. Each of these cell monitoring strategies evaluate the recognition of the APC by the CTL. It will be desirable to use as antigen presenting cells, cells that have been either infected, or transfected with the appropriate genes, or, (generally only for class II epitopes, due to the different peptide processing pathway for HLA class 11), cells that have been pulsed with whole protein antigens, to establish whether endogenously produced antigen is also recognized by the T cells induced by the analog peptide. It is to be noted that peptide/protein-pulsed dendritic cells can be used to present whole protein antigens for both HLA class I and class II.
[0245] Another embodiment of the invention is to create analogs of weak binding peptides, to thereby ensure adequate numbers of cellular binders. Class I
binding peptides exhibiting binding affinities of 500-5000 nM, and carrying an acceptable but suboptimal primary anchor residue at one or both positions can be "fixed"
by substituting preferred anchor residues in accordance with the respective supertype. The analog peptides can then be tested for binding and/or cross-binding capacity.
[0246] Another embodiment of the invention is to create analogs of peptides that are already cross-reactive binders and are vaccine candidates, but which bind weakly to one or more alleles of a supertype. If the cross-reactive binder carries a suboptimal residue (less preferred or deleterious) at a primary or secondary anchor position, the peptide can be analoged by substituting out a deleterious residue and replacing it with a preferred or less preferred one, or by substituting out a less preferred residue and replacing it with a preferred one. The analog peptide can then be tested for cross-binding capacity.
[0247] Another embodiment for generating effective peptide analogs involves the substitution of residues that have an adverse impact on peptide stability or solubility in, e.g., a liquid environment. This substitution may occur at any position of the peptide epitope. For example, a cysteine (C) can be substituted out in favor of a-amino butyric acid. Due to its chemical nature, cysteine has the propensity to form disulfide bridges and sufficiently alter the peptide structurally so as to reduce binding capacity. Substituting a-amino butyric acid for C not only alleviates this problem, but actually improves binding and crossbinding capability in certain instances (see, e.g., the review by Sette et al., In: Persistent Viral Infections, Eds. R. Ahmed and I. Chen, John Wiley & Sons, England, 1999).
Substitution of cysteine with a-amino butyric acid may occur at any residue of a peptide epitope, i.e. at either anchor or non-anchor positions.

[024] Moreover, it has been shown that in sets of A*0201 motif-bearing peptides containing at least one preferred secondary anchor residue while avoiding the presence of any deleterious secondary anchor residues, 69% of the peptides will bind A*0201 with an IC50 less than 500 nM (Ruppert, J. et al.
Cell 74:929, 1993). The determination of what was a preferred or deleterious residue in Ruppert can be used to generate algorithms (see, e.g., 22). Such algorithms are flexible in that cut-off scores may be adjusted to select sets of peptides with greater or lower predicted binding properties, as desired.
[0249] C35 epiotpes containing cysteine residues have a tendency to dimerize with other cysteine containing peptides. Thus, an embodiment of the present invention is a composition comprising a peptide epitope of the invention (e.g.
a C35 peptide epitope listed in any of Tables 1-3 or 5-6, exclusive of E100 to of SEQ ID NO:2) and a suitable reducing agent that protects the free sulthydryl group of the cysteine residue but does not otherwise inhibit epitope binding.
In a preferred embodiment the composition comprises the peptide epitope ITNSRPPCV or KITNSRPPCV in combination with a suitable reducing agent.
Suitable reducing agents include, but are not limited to, TCEP and dithiothreitol (DTT).
[0250] Another embodiment of the invention is to create peptide epitope analogs in which the cysteine residues of the peptide epitope (e.g., a C35 peptide epitope listed in any of Tables 1-3 or 5-6, exclusive of E100 to R109 of SEQ ID NO:2) have been substituted with any other amino acid to facilitate synthesis. (See Zarling, A.L. et al., J. Exp. Med. 192(12): 1755-1762 (2000)).

Preferably, the cysteine residues are substituted with either alanine, serine or glycine residues, although any amino acid can be substituted provided that such substitution does not negatively effect binding to MHC or recognition by T cells. Thus, in a particularly preferred embodiment, the isolated polypeptides of the present invention comprise or, alternatively, consist of the following C35 peptide epitope analogs: for the peptide epitope G22 to C30 of SEQ ID
NO:2 and FIG. 1B (i.e., GVRIVVEYC), the analog with either alanine or glycine substituted for the cysteine at the ninth amino acid residue (i.e., GVRIVVEYA or GVRIVVEYQ); for the peptide epitope 125 to C33 of SEQ
ID NO:2 and FIG. 1B (i.e., IVVEYCEPC), the analog with either alanthe or glycine substituted for the cysteine at the sixth amino acid residue and/or the ninth amino acid residue (i.e., TVVEYAEPC or IVVEYGEPC or TVVEYCEPA or IVVEYCEPG or IVVEYAEPA or TVVEYAEPG or IVVEYGEPA or IVVEYGEPG); for the peptide epitope of K104 to C112 of SEQ ID NO:2 and FIG. 1B (i.e., KITNSRPPC) the analog with either alanine or glycine substituted for the cysteine at the ninth residue (i.e., KITNSRPPA
or K1TNSRPPg); for the peptide epitope K104 to V113 of SEQ ID NO:2 and FIG. 1B (i.e., KITNSRPPCV), the analog with either alanine, serine or glycine substituted for the cysteine at the ninth residue (i.e., KITNSRPPAV, KITNSRPPSV or KITNSRPPGV); for the peptide epitope 1105 to V113 of SEQ ID NO:2 and FIG. 1B (i.e., ITNSRPPCV), the analog with either alanine, serine or glycine substituted for the cysteine at the eighth residue (i.e., ITNSRPPAV, ITNSRPPSV or ITNSRPPGV); for the peptide epitope N107 to L115 (i.e., NSRPPCVIL), the analog with either alanine or glycine substituted for the cysteine at the sixth amino acid residue (i.e., NSRPPAVIL, NSRPPGVIL); for the multi-epitope peptide T101 to V113 of SEQ ID NO:2 and FIG. 1B (i.e., TLEK1TNSRPPCV), the analog with either alanine or glycine substituted for the cysteine at the twelfth residue (i.e., TLEKITNSRPPAV or TLEKITNSRPPGV); for the multi-epitope peptide E100 to V113 of SEQ ID NO:2 and FIG. 1B (i.e., ETLEKITNSRPPCV), the analog with either alanine or glycine substituted for the cysteine at the thirteenth amino acid residue (i.e., ETLEKITNSRPPAV, ETLEKITNSRPPGV), for the multi-epitope peptide G99 to V113 of SEQ ID
NO:2 and FIG. 1B (i.e., GETLEKITNSRPPCV), the analog with either alanine or glycine substituted for the cysteine at the fourteenth amino acid residue (i.e., GETLEKITNSRPPAV, GETLEKITNSRPPGV), for the multi-epitope peptide 193 to V113 of SEQ ID NO:2 and FIG. 1B (i.e., IRRASNGETLEKITNSRPPCV), the analog with either alanine or glycine substituted for the cysteine at the twentieth residue (i.e., IRRASNGETLEKITNSRPPAV or IRRASNGETLEKITNSRPPGV); for the multi-epitope peptide D88 to V113 of SEQ ID NO:2 and FIG. 1B (i.e., DLIEAIRRASNGETLEKTINSRPPCV), the analog with either alanine or glycine substituted for the cysteine at the twenty-fifth residue (i.e., DLIEAIRRASNGETLEKITNSRPPAV or DLIEAIRRASNGETLEKITNSRPPGV); for the multi-epitope peptide P84 to V113 of SEQ ID NO:2 and FIG. 1B (i.e., PYEKDLIEAIRRASNGETLEKITNSRPPCV), the analog with either alanine or glycine substituted for the cysteine at the twenty-ninth residue (i.e., PYEKDLIEAIRRASNGETLEKITNSRPPAV or PYEKDLIEAIRRASNGETLEKITNSRPPGV); for the multi-epitope peptide K77 to L115 of SEQ ID NO:2 and FIG. 1B (i.e., KLENGGFPYEKDLIEAIRRASNGETLEKITNSRPPCV), the analog with either alanine or glycine substituted for the cysteine at the thirty-sixth residue (i.e., KLENGGFPYEKDLIEAIRRASNGETLEKITNSRPPAV or KLENGGFPYEKDLIEAIRRASNGETLEKITNSRPPGV); for the multi-epitope peptide Q72 to L115of SEQ ID NO:2 and FIG. 1B (i.e., QLVFSKLENGGFPYEKDLIEAIRRASNGETLEKITNSRPPCV), the analog with either alanine or glycine substituted for the cysteine at the forty-first residue (i.e., QLVFSKLENGGFPYEKDLIEAIRRASNGETLEKITNSRPPAV
or QLVFSKLENGGFPYEKDLIEAIRRASNGETLEKITNSRPPGV); for the multi-epitope peptide F65 to L115 of SEQ ID NO:2 and FIG. 1B (i.e., FEIEINGQLVFSKLENGGFPYEKDL1EAIRRASNGETLEKITNSRPPCV), the analog with either alanine or glycine substituted for the cysteine at the forty-eighth residue (i.e., FELEINGQLVFSKLENGGFPYEKDLIEAIRRASNGETLEKITNSRPPAV or FEIEINGQLVFSKLENGGFPYEKDLIEAIRRASNGETLEKITNSRPPGV);
and for the multi-epitope peptide L59 to L115 of SEQ ID NO:2 and FIG. 1B

(i.e., LGGTGAFEIEINGQLVFSKLENGGFPYEKDLIEAIRRASNGETLEKITNS
RPPCV), the analog with either alanine or glycine substituted for the cysteine at the fifty-fourth residue (i.e., LGGTGAFEIEINGQLVFSKLENGGFPYEKDLIEAIRRASNGETLEKTINS
RPPAV or LGGTGAFEMINGQLVFSKLENGGFPYEKDLIEAIRRASNGETLEKITNS
RPPGV) [0251] Another embodiment of the invention is to create peptide epitope analogs in which the cysteine residues of the peptide epitope (e.g., a C35 peptide epitope listed in any of Tables 1-3 or 5-6, exclusive of E100 to R109 of SEQ ID NO:2, having one or more cysteine residues) have been "cysteinylated" (i.e., reacted with a second cysteine residue). (See Pierce, R.A. et al., J. Immunol.
163(12):6360-6364 (1999)). As used herein, the term "cysteinylated" describes a cysteine residue, within a peptide (e.g. peptide epitope) of the present invention, which has been reacted with a second free cysteine, i.e. a cysteine not part of a larger peptide, at the free sulfhydryl group thereby creating a disulfide bond (-SH
+ HS- = -S-S-).
[0252] Thus, in a particularly preferred embodiment, the isolated polyp eptides of the present invention comprise or, alternatively, consist of the following C35 peptide epitope analogs: for the peptide epitope of K104 to V113 of SEQ ID
NO:2 and FIG. 1B (i.e., KITNSRPPCV), the analog wherein the cysteine at the ninth residue has been cysteinylated and for the peptide epitope of 11 05 to of SEQ ID NO:2 and FIG. 1B (i.e. ITNSRPPCV), the analog wherein the cysteine at the eighth residue has been cysteinylated.
[0253] Another embodiment of the invention is to create peptide epitope analogs in which the serine, threonine and/or tyrosine residues of the peptide epitope (e.g., a C35 peptide epitope listed in any of Tables 1-3 or 5-6, exclusive of E100 to R109 of SEQ ID NO:2) have been phosphorylated. Thus, in a particularly preferred embodiment, the isolated polypeptides of the present invention comprise or, alternatively, consist of the following C35 peptide epitope analogs: for the peptide epitope E4 to P12 of SEQ ID NO:2 and FIG.
1B (i.e., EPGQTSVAP), the analog wherein the threonine at T8 and/or the serine at S9 have been phosphorylated; for the peptide epitope S9 to V17 of SEQ ID NO:2 and FIG. 1B (i.e., SVAPPPEEV), the analog wherein the serine at S9 has been phosporylated; for the peptide epitope 521 to Y29 of SEQ ID
NO:2 and FIG. 1B (i.e., SGVRIVVEY), the analog wherein the serine at S21 and/or the tyrosine at Y29 are phosphorylated; for the peptide epitope G22 to C30 of SEQ ID NO:2 and FIG. 1B (i.e., GVRIVVEYC), the analog wherein the tyrosine at Y29 is phosphorylated; for the peptide epitope T38 to V46 of SEQ ID NO:2 and FIG. 1B (i.e., TYLELASAV), the analog wherein the threonine at T38, the tyrosine at Y39, and/or the serine at S44 are phosphorylated; for the peptide epitope G61 to 169 (i.e., GTGAFEIEI), the analog wherein the threonine at T62 is phosphorylated); for the peptide epitope T62 to N70 of SEQ ID NO:2 and FIG. 1B (i.e., TGAFEIEIN), the analog wherein the threonine at T62 has been phosphorylated; for the peptide epitope K77 to Y85 of SEQ ID NO:2 and FIG. 1B (i.e., KLENGGFPY), the analog wherein the tyrosine at Y85 is phosphorylated; for the peptide epitope Q72 to E86 of SEQ ID NO:2 and FIG. 1B (i.e., QLVFSKLENGGFPYE), the analog wherein the serine at S76 and/or the tyrosine at Y85 are phosphorylated; for the peptide epitope G81 to L89 of SEQ ID NO:2 or FIG. 1B (i.e., GGFPYEKDL), the analog wherein the tyrosine at Y85 is phosphorylated; for the peptide epitope K104 to C112 of SEQ ID NO:2 and FIG. 1B (i.e., KITNSRPPC), the analog wherein the threonine at T106 and/or the serine at S108 are phosphorylated; for the peptide epitope K104 to V113 of SEQ ID
NO:2 and FIG. 1B (i.e., KITNSRPPCV), the analog wherein the threonine at T106 and/or the serine at S108 are phosphorylated; for the peptide epitope 1105 to V113 (i.e., ITNSRPPCV), the analog wherein the threonine at T106 and/or the serine at S108 are phosphorylated; for the peptide epitope N107 to L115 (i.e., NSRPPCVIL), the analog wherein the serine at S108 is phosphorylated; for the polyepitopic peptide T101 to V113 of SEQ ID NO:2 and FIG. 1B (i.e., TLEKITNSRPPCV), the analog wherein the threonines at T101 and T106 and/or the serine at S108 are phosphorylated; for the polyepitopic peptide 193 to V113 of SEQ ID NO:2 and FIG. 1B (i.e., MRASNGETLEKITNSRPPCV), the analog wherein the serine at S97 and/or the threonine at T101 and/or the threonine at T106 and/or the serine at S108 are phosphorylated; for the polyepitopic peptide D88 to V113 of SEQ ID NO:2 and FIG. 1B (i.e., DLIEAlRRASNGETLEKITNSRPPCV), the analog wherein the serine at S97 and/or the threonine at T101 and/or the threonine at T106 and/or the serine at S108 are phosphorylated; for the polyepitopic peptide P84 to V113 of SEQ ID NO:2 and FIG. 1B (i.e., PYEKDLIEAlRRASNGETLEKITNSRPPCV), the analog wherein the tyrosine at Y85 and/or the serine at S97 and/or the threonine at T101 and/or the threonine at T106 and/or the serine at S108 are phosphorylated; for the polyepitopic peptide K77 to V113 of SEQ ID NO:2 and FIG. 1B (i.e., KLENGGFPYEKDLFEAIRRASNGETLEKITNSRPPCV), the analog wherein the tyrosine at Y85 and/or the serine at S97 and/or the threonine at T101 and/or the threonine at T106 and/or the serine at S108 are phosphorylated; for the polyepitopic peptide Q72 to V113 of SEQ ID NO:2 and FIG. 1B (i.e., QLVFSKLENGGFPYEKDLIEAIRRASNGETLEKITNSRPPCV), the analog wherein the serine at S76 and/or the tyrosine at Y85 and/or the serine at S97 and/or the threonine at T101 and/or the threonine at T106 and/or the serine at S108 are phosphorylated; for the polyepitopic peptide F65 to V113 of SEQ ID
NO:2 and FIG. 1B (i.e., FEIONGQLVFSKLENGGFPYEKDLIEAlRRASNGETLEKITNSRPPCV), the analog wherein the serine at S76 and/or the tyrosine at Y85 and/or the serine at S97 and/or the threonine at T101 and/or the threonine at T106 and/or the serine at S108 are phosphorylated; for the polyepitopic peptide L59 to V113 of SEQ ID NO:2 and FIG. 1B (i.e., LGGTGAFEIElNGQLVFSKLENGGFPYEKDLIEA1RRASNGETLEKITNS

RPPCV), the analog wherein the threonine at T62 and/or the serine at S76 and/or the tyrosine at Y85 and/or the serine at S97 and/or the threonine at T101 and/or the threonine at T106 and/or the serine at S108 are phosphorylated.
[0254] Another embodiment of the invention is to create peptide epitope analogs in which the asparagine residues of the peptide epitope (e.g., a C35 peptide epitope listed in any of Tables 1-3 or 5-6, exclusive of E100 to R109 of SEQ ID NO:2) have been converted to aspartic acid after translation. (See Skipper, J.C. et al., J. Exp. Med. 183(2):527-534 (1996)).
[0255] In preferred embodiments, the C35 peptide epitope analogs of the present invention contain multiple modifications provided that such modifications do not inhibit binding to MHC molecules or recognition by T
cells. Thus, preferred analogs include C35 peptide epitopes for which one or more residues have been modified as described herein to increase binding affinity to MHC molecules, one or more cysteine residues have been replaced with alanine or glycine residues to facilitate synthesis, and one or more serine, threonine or tyrosine residues have been phosphorylated.
[0256] Furthermore, additional amino acids can be added to the termini of a peptide epitope to provide for ease of linking peptide epitopes one to another, for coupling to a carrier support or larger polypeptide, for modifying the physical or chemical properties of the peptide or oligopeptide, or the like.
Amino acids such as tyrosine, cysteine, lysine, glutamic or aspartic acid, or the like, can be introduced at the C- or N-terminus of the peptide or oligopeptide, particularly class I peptides. It is to be noted that modification at the carboxyl terminus of a CTL epitope may, in some cases, alter binding characteristics of the peptide. In addition, the peptide or oligopeptide sequences can differ from the natural sequence by being modified by terminal-NH2 acylation, e.g., by alkanoyl (C1-C20) or thioglycolyl acetylation, terminal-carboxyl amidation, e.g., ammonia, methylamine, etc., polyethylene-glycol modification (i.e., PEGylation) of the C-terminus, and the addition of a lipid tail (e.g., a palmitoyl-lysine chain) to enhance presentation to T cells and immunogenicity.

(See Brinckerhoff, L.H. et al., Int. J. Cancer 83(3):326-334 (1999); Le Gal, F.A. et al., Int. J. Cancer 98(2):221-227 (2002). N-terminal amides, in particular, will be more resistant to certain peptidases, thus preventing destruction of the peptide epitope in situ without affecting recognition. This will effectively increase the half-life of the peptide epitope and enhance its ability to stimulate immune cells. In some instances these modifications may provide sites for linking to a support or other molecule.
Preparation of Peptide Epitopes [0257] Peptide epitopes in accordance with the invention can be prepared synthetically, by recombinant DNA technology or chemical synthesis, or from natural sources such as native tumors or pathogenic organisms. Peptide epitopes may be synthesized individually or as polyepitopic polypeptides (e.g., homopolymers or heteropolymers). Although the peptide will preferably be substantially free of other naturally occurring host cell proteins and fragments thereof, in some embodiments the peptides may be synthetically conjugated to native fragments or particles.
[0258] In addition, one or more non-C35 tumor associated peptides can be linked to one or more C35 peptide epitopes and/or C35 peptide epitope analogs to increase immune response via HLA class I and/or class II.
Especially preferred are polypeptides comprising a series of epitopes, known as "polytopes," and nucleic acids encoding same. The epitopes can be arranged in sequential or overlapping fashion (see, e.g., Thomson et al., Proc.
Natl. Acad. ScL USA 92:5845-5849 (1995); Gilbert et al., Nature/Biotechnology 15:1280-1284 (1997)), with or without the natural flanking sequences, and can be separated by unrelated linker sequences if desired. The polytope is processed to generate individual epitopes which are recognized by the immune system for generation of immune responses.

[0259] Thus, for example, C35 peptide epitopes and C35 peptide epitope analogs can be combined with peptides from other tumor rejection antigens (e.g., by preparation of hybrid nucleic acids or polypeptides) to form "polytopes." (Zeng, G. et al., Proc. Natl. Acad. Sci. 98(7):3964-3969 (2001);
Zeng, G. et al., J. Immunol. 165:1153-1159 (2000); Mancini, S. et al., J. Exp.

Med. 189(5):871-876 (1999)). Exemplary tumor associated antigens that can be administered to induce or enhance an immune response are derived from tumor associated genes and encoded proteins including: MAGE-1, MAGE-2, MAGE-3, MAGE-4, MAGE-5, MAGE-6, MAGE-7, MAGE-8, MAGE-9, MAGE-10, MAGE-11, MAGE-12, MAGE-13, GAGE-1, GAGE-2, GAGE-3, GAGE-4, GAGE-5, GAGE-6, GAGE-7, GAGE-8, BAGE-1, RAGE-1, LB33/MUM-1, PRAME, NAG, MAGE-Xp2 (MAGE-B2), MAGE-Xp3 (MAGE-B3), MAGE-Xp4 (MAGE-B4), tyrosinase, brain glycogen phosphorylase, Melan-A, MAGE-C1, MAGE-C2, NY-ES0-1, LAGE-1, SSX-1, SSX-2(HOM-MEL-40), SSX-1, SSX-4, SSX-5, SCP-1 and CT-7. For example, specific antigenic peptides characteristic of tumors include those listed in Table A.
Table A
Gene 111HC Peptide Position MAGE-1 HLA-Al EADPTGHSY 161-169 HLA-Cw16 SAYGEPRKL 230-238 MAGE-3 HLA-Al EVDPIGHLY 168-176 BAGE HLA-Cw16 AARA'VFLAL 2-10 GAGE-1,2 HLA-Cw16 YRPRPRRY 9-16 GnT-V 1{LA-A2 VLPDVFIRC(V) 2-10/11 MUM-1 HLA-B44 EEKLIVVLF exon 2/intron EEKLSVVLF (wild-type) ARDPHSGHFV (wild-type) 13-catenin HLA-A24 SYLDSGIHF 29-37 SYLDSGIHS (wild-type) Tyrosinase HLA-A2 MLLA'VLYCL 1-9 Melan-Amartl HLA-A2 (E)AAGIGILTV 26/27-35 gp1001'117 HLA-A2 KTWGQYWQV 154-162 MAGE-6 HLA-Cw16 KISGGPRISYPL 292-303 [0260] Other examples of non-C35 HLA class I and HLA class II binding peptides will be known to one of ordinary skill in the art and can be used in the invention in a like manner to those disclosed herein. One of ordinary skill in the art can prepare polypeptides comprising one or more C35 peptide epitopes or peptide epitope analogs and one or more of the aforementioned tumor rejection peptides, or nucleic acids encoding such polypeptides, according to standard procedures in molecular biology. Examples ofpolytopes comprising C35 peptide epitopes or C35 peptide epitope analogs of the present invention and various tumor rejection antigenic peptides are set forth in Tables B and C below.
[0261] Thus, polytopes are groups of two or more potentially immunogenic or immune response stimulating peptides which can be joined together in various arrangements (e.g. concatenated, overlapping). The polytope (or nucleic acid encoding the polytope) can be administered in a standard immunization protocol, e.g. to animals, to test the effectiveness of the polytope in stimulating, enhancing and/or provoking an immune response. The peptides can be joined directly or via the use of flanking sequences to form polytopes, and the use of polytopes as vaccines is well known in the art.
[0262] In a preferred embodiment, the isolated polypeptides of the present invention comprise one or more C35 peptide epitopes or C35 peptide epitope analogs linked to one or more tumor rejection peptides. In a particularlypreferred embodiment, said one or more C35 peptide epitopes are selected from the group consisting of: amino acids E4 to P12 of SEQ ID NO:2, amino acids S9 to V17 of SEQ ID NO:2, amino acids S21 to Y29 of SEQ ID NO:2, amino acids G22 to C30 of SEQ ID NO: 2, amino acids 125 to C33 of SEQ ID NO:2, amino acids T38 to V46 of SEQ ID NO:2, amino acids G61 to 169 of SEQ ID NO:2, amino acids T62 to N70 of SEQ ID NO:2, amino acids G63 to G71 of SEQ ID NO:2, amino acids F65 to L73 of SEQ ID NO:2, amino acids 167 to F75 of SEQ ID NO:2, amino acids K77 to Y85 of SEQ ID NO:2, amino acids Q72 to E86 of SEQ ID
NO:2, amino acids G81 to L89 of SEQ ID NO:2, amino acids K104 to C112 of SEQ ID NO:2, amino acids K104 to V113 of SEQ ID NO:2, amino acids 1105 to V113 of SEQ ID NO:2, amino acids N107 to L115 of SEQ ID NO:2, amino acids T101 to V113 of SEQ ID NO:2, amino acids E100 to V113 of SEQ ID
NO:2, amino acids G99 to V113 of SEQ ID NO:2, amino acids 193 to V113 of SEQ ID NO:2, amino acids D88 to V113 of SEQ ED NO:2, amino acids P84 to V113 of SEQ NO:2, amino acids K77 to V113 of SEQ ID NO:2, amino acids Q72 to V113 of SEQ ID NO:2, amino acids F65 to V113 of SEQ ID NO:2, and L57 to V113 of SEQ ID NO:2; and said one or more tumor rejection peptides are selected from the group consisting of the antigenic peptides shown in Table A.
[0263] In another embodiment, one or more non-C35 cell penetrating peptides can be linked to one or more C35 peptide epitopes and/or C35 peptide epitope analogs to enhance delivery of C35 peptide epitopes to cells, e.g., dendritic cells.
Especially preferred are polypeptides comprising a series of C35 peptide epitopes or C35 peptide epitope analogs and cell penetrating peptides, and nucleic acids encoding same. The epitopes and peptides can be arranged in sequential or overlapping fashion with or without the natural flanking sequences, and can be separated by unrelated linker sequences if desired. The polypeptide is processed to generate individual C35 epitopes which are recognized by the immune system for generation of immune responses.
[0264] Thus, for example, C35 peptide epitopes and C35 peptide epitope analogs can be combined with cell-penetrating peptides. (Wang, R.-F. et al., Nature Biotechnology 20(2):149-154 (2002); Frankel, A.D. et al., Cell 55:1189-1193 (1988); Elliott, G. et al., Cell 88(2):223-233 (1997); Phelan, A. et al., Nature Biotechnology 16(5):440-443 (1998); Lin, Y.-Z. et al., J. Biol. Chem.
270(24):14255-14258 (1995); Rojas, M. et al., Nature Biotechnology 16(4):370-375 (1998)). Exemplary cell penetrating peptides that can be administered to enhance delivery of C35 peptides to cells, such as denthitic cells, include:
the Tat protein of human immunodeficiency virus, the HSV-1 structural protein VP22, and the 12-residue membrane-translocating sequence (MTS) modified from the 16-residue h region of the signal sequence of Kaposi fibroblast growth factor.
[0265] The one or more C35 peptide epitopes/analogs and one or more cell penetrating peptides can be joined together in various arrangements (e.g.
concatenated, overlapping). The resulting polypeptide (or nucleic acid encoding the polypeptide) can be administered in a standard immunization protocol, e.g.
to animals, to test the effectiveness of the polypeptide in stimulating, enhancing and/or provoking an immune response. The C35 peptide epitopes/analogs and one or more cell penetrating peptides can be joined directly or via the use of flanking sequences to form the polypeptides, and the use of such polypeptides as vaccines is well known in the art. Examples of polypeptides comprising C35 peptide epitopes or C35 peptide epitope analogs of the present invention and various cell penetrating peptides are set forth in Tables D and E below.

TABLE B

C35 Peptide/Epitope Exemplary Tumor Rejection Peptide Exemplary Polytopes S9 -V17 of SEQ ID NO:2 amino acids 161-169 of MAGE-1 SVAPPPEEVEADPTGHSY, tµ.) oe SVAPPPEEV EADPTGHSYSVAAPPPEEVEADPTGHSY
amino acids 230-238 of MAGE-1 SVAPPPEEVSAYGEPRKL, SAYGEPRKL SVAPPPEEVSAYGEPRKL
amino acids 168-176 of MAGE-3 SVAPPPEEVEVDPIGHLY, EVDPIGHLYSVAPPPEEVEVDPIGHLY
amino acids 271-279 of MAGE-3 SVAPPPEEVFLWGPRALV, FLWGPRALVSVAPPPEEVFLWGPRALV

amino acids 167-176 of MAGE-3 SVAPPPEEVMEVDPIGHLY, co co c7, MEVDPIGHLYSVAPPPEEVMEVDPIGHLY
co 1\.) amino acids 2-10 of BAGE SVAPPPEEVAARAVFLAL, AARAVFLALSVAPPPEEVAARAVFLAL
amino acids 9-16 of GAGE-1,2 SVAPPPEEVYRPRPRRY, YRPRPRRYSVAPPPEEVYRPRPRRY
c7, amino acids 11-20 of RAGE SVAPPPEEVSPSSNRIRNT, SPSSNRIRNTSVAPPPEEVSPSSNRIRNT
amino acids 23-32 of CDK4 SVAPPPEEVARDPHSGHFV, ARDPHSGHFVSVAPPPEEVARDPHSGHFV
amino acids 29-37 of P-catenin SVAPPPEEVSYLDSGIHS, SYLDSGHISSVAPPPEEVSYLDSGIHS

amino acids 1-9 of Tyrosinase SVAPPPEEVMLLAVLYCL, MLLAVLYCLSVAPPPEEVMLLAVLYCL
oe tµ.) c.;11 tµ.) C35 Peptide/Epitope Exemplary Tumor Rejection Peptide Exemplary Polytopes amino acids 206-214 of Tyrosinase SVAPPPEEVAFLPWHRLF, AFLPWHRLFSVAPPPEEVAFLPWHRLF
amino acids 56-70 of Tyrosinase SVAPPPEEVQNILLSNAPLGPQFP, QNILLSNAPLGPQFP SVAPPPEEVQNILLSNAPLGPQFP
amino acids 448-462 of Tyrosinase SVAPPPEEVDYSYLQDSDPDSFQD, DYSYLQDSDPDSFQDSVAPPPEEVDYSYLQDSDPDSFQD
amino acids 32-40 of Melan-AmART-1 SVAPPPEEVJLTVILGVL, JLTVILGVLSVAPPPEEVJLTVILGVL
amino acids 154-162 of gpl00Pme117 SVAPPPEEVKTWGQYWQV, KTWGQYVVQVSVAPPPEEVKTWGQYWQV

amino acids 209-217 of gp100'117 SVAPPPEEVITDQVPFSV, c7, op ITDQVPFSVSVAPPPEEVITDQVPFSV
"N) amino acids 280-288 of gp100'117 SVAPPPEEVYLEPGPVTA, YLEPGPVTASVAPPPEEVYLEPGPVTA
amino acids 457-466 of gp1001'117 SVAPPPEEVLLDGTATLRL, c7, LLDGTATLRLSVAPPPEEVLLDGTATLRL
amino acids 476-485 of gp100"117 SVAPPPEEVVLYRYGSFSV, VLYRYGSFSVSVAPPPEEVVLYRYGSFSV
amino acids 301-309 of PRAME SVAPPPEEVLYVDSLFFL, LYVDSLFELSVAPPPEEVLYVDSLEFL
amino acids 292-303 of MAGE-6 SVAPPPEEVKISGGPRISYPL, KIS GGPRISYPLSVAPPPEEVKIS GGPRISYPL
amino acids 157-167 of NY-ESO-1 SVAPPPEEVSLLMWITQCFL, SLLMWITQCFLSVAPPPEEVSLLMWITQCFL

C35 Peptide/Epitope Exemplary Tumor Rejection Peptide Exemplary Polytopes amino acids 157-165 of NY-ESO-1 SVAPPPEEVSLLMWITQC, SLLMWITQCSVAPPPEEVSLLMWITQC
tµ.) amino acids 155-163 of NY-ESO-1 SVAPPPEEVQLSLLMWIT, oe QLSLLMWITSVAPPPEEVQLSLLMWIT
amino acids 157-170 of NY-ESO-1 SLLMWITQCFLPVFSVAPPPEEVTSYVKVLHHMVKISG
and amino acids 281-295 of MAGE-3 S21 -Y29 of SEQ NO:2 amino acids 161-169 of MAGE-1 SGVRIVVEYEADPTGHSY, SGVRIVVEY EADPTGHSYSGVRIVVEYEADPTGHSY
amino acids 230-238 of MAGE-1 SGVRIVVEYSAYGEPRKL, SAYGEPRKLSGVRIVVEYSAYGEPRKL
co co amino acids 168-176 of MAGE-3 SGVRIVVEYEVDPIGHLY, c7, co EVDPIGHLYSGVRIVVEYEVDPIGHLY
1\) amino acids 271-279 of MAGE-3 SGVRIVVEYFLWGPRALV, FLWGPRALVSGVRIVVEYFLWGPRALV
amino acids 167-176 of MAGE-3 SGVRIVVEYMEVDPIGHLY, c7, MEVDPIGHLYSGVRIVVEYMEVDPIGHLY
amino acids 2-10 of BAGE SGVRIVVEYAARAVFLAL, AARAVFLALSGVRIVVEYAARAVFLAL
amino acids 9-16 of GAGE-1,2 SGVRIVVEYYRPRPRRY, YRPRPRRYSGVRIVVEYYRPRPRRY
amino acids 11-20 of RAGE SGVRIVVEYSPSSNRIRNT, SPSSNRIRNTSGVRIVVEYSPSSNRIRNT
amino acids 23-32 of CDK4 SGVRTVVEYACDPHSGHFV, ACDPHSGHFVSGVRIVVEYACDPHSGHFV
tµ.) tµ.) C35 Peptide/Epitope Exemplary Tumor Rejection Peptide Exemplary Polytopes amino acids 29-37 of 0-catenin S GVRIVVEYSYLD S GIHF, SYLD S GIEFS GVRIVVEYSYLD SGIHF
tµ.) amino acids 1-9 of tyrosinase SGVRIV VEYMLLAVLYCL, MLLAVLYCLS GVRIVVEYMLLAVLYCL
amino acids 206-214 of tyrosinase S GVRIVVEYAFLPWHRLF, AFLPWHRLFS GVRIVVEYAFLPWHRLF
amino acids 56-70 of tyrosinase S GVRIVVEYQNILLSNAPLGPQFP, QNILLSNAPLGPQFP S GVRIVVEYQNILLSNAPLGPQFP
amino acids 448-462 of tyrosinase SGVRIVVEYDYSYLQD SDPD SFQD, DYSYLQD SDPD SFQD S GVRIVVEYDYSYLQD SDPD SFQD

amino acids 32-40 of Me1an-AmART-1 SGVRIVVEYJLTVILGVL, c7, JLTVILGVLSGVRIVVEYJLTVILGVL
amino acids 154-162 of gplOOP'117 SGVRIVVEYKTWGQYWQV, KTWGQYWQVS GVRIVVEYKTWGQYWQV
amino acids 209-217 of gplOOPmel" SGVRIVVEY ITDQVPFSV, c7, ITDQVPFSV SGV'RIVVEY ITDQVPFSV
amino acids 280-288 of gp100'117 S GVRIVVEYYLEP GPVTA, YLEPGPVTASGVRIVVEYYLEPGPVTA
amino acids 457-466 of gp100' 17 SGVRIVVEYLLDGTATLRL, LLD GTATLRLS GVRIVVEYLLD GTATLRL
amino acids 476-485 of gp100Pmel 17 SGVRIVVEYVLYRYGSFSV, VLYRYGSFSVS GVRIVVEYVLYRYGSFSV
amino acids 301-309 of PRAME S GVRIVVEYLYVD SLFFL, LYVD SLFFLS GVRIVVEYLYVD SLFFL
oe JI
tµ.) tµ.) . =

C35 Peptide/Epitope Exemplary Tumor Rejection Peptide Exemplary Polytopes amino acids 292-303 of MAGE-6 SGVRIVVEYKISGGPRISYPL, KISGGPRISYPLSGVRIVVEYKISGGPRISYPL
amino acids 157-167 of NY-ESO-1 SGVRIVVEYSLLMWITQCFL, tµ.) SLLMWITQCFLSGVRIVVEYSLLMWITQCFL
amino acids 157-165 of NY-ESO-1 SGVRIVVEYSLLMWITQC, SLLMWITQCSGVRIVVEYSLLMWITQC
amino acids 155-163 of NY-ESO-1 SGVRIVVEYQLSLLMWIT, QLSLLMWITSGVRIVVEYQLSLLMWIT
amino acids 157-170 of NY-ESO-1 SLLMWITQCFLPVFSGVRIVVEYTSYVKVLHHMVKISG
and amino acids 281-295 of MAGE-3 co G22-C30 of SEQ ID NO:2 amino acids 161-169 of MAGE-1 GVRIVVEYCEADPTGHSY, uco GVRIVVEYC EADPTGHSYGVRIVVEYCEADPTGHSY
r.,4 "
amino acids 230-238 of MAGE-1 GVR1VVEYCSAYGEPRKL, SAYGEPRKLGVRIVVEYCSAYGEPRKL
amino acids 168-176 of MAGE-3 GVRIVVEYCEVDPIGHLY, EVDPIGHLYGVRIVVEYCEVDPIGHLY
c7, amino acids 271-279 of MAGE-3 GVRIVVEYCFLWGPRALV, FLWGPRALVGVRIVVEYCFLWGPRALV
amino acids 167-176 of MAGE-3 GVRIVVEYCMEVDPIGHLY, MEVDPIGHLYGVRIVVEYCMEVDPIGHLY
amino acids 2-10 of BAGE GVRIVVEYCAARAVFLAL, amino acids 9-16 of GAGE-1,2 GVRIVVEYCYRPRPRRY, YRPRPRRYGVRIVVEYCYRPRPRRY
oe tµ.) tµ.) C35 Peptide/Epitope Exemplary Tumor Rejection Peptide Exemplary Polytopes amino acids 11-20 of RAGE GVRIVVEYCSPSSNRJR_NT, SPSSNRIRNTGVRIVVEYCSPSSNRIRNT
tµ.) amino acids 23-32 of CDK4 GVRIVVEYCACDPHSGHFV, ACDPHSGHFVGVRIVVEYCACDPHSGHFV
amino acids 29-37 of P-catenin GVRIVVEYCSYLDSGIHF, SYLDSGIHFGVRIVVEYCSYLDSGIHF
amino acids 1-9 of tyrosinase GVRIVVEYCMLLAVLYCL, MLLAVLYCLGVRIVVEYCMLLAVLYCL
amino acids 206-214 of tyrosinase GVRIVVEYCAFLPWHRLF, AFLPWHRLFGVRIVVEYCAFLPWHRLF

co amino acids 56-70 of tyrosinase GVRIVVEYCQNILLSNAPLGPQFP, uco c7, QNILLSNAPLGPQFPGVRIVVEYCQNILLSNAPLGPQFP
amino acids 448-462 of tyrosinase GVRIVVEYCDYSYLQDSDPDSFQD, DYSYLQDSDPDSFQDGVRIVVEYCDYSYLQDSDPDSFQD
amino acids 32-40 of Melan-A' GVRIVVEYCJLTVILGVL, c7, JLTVILGVLGVRIVVEYCJLTVILGVL
amino acids 154-162 of gplOOP'117 GVRIVVEYCKTWGQYWQV, KTWGQYWQVGVRIVVEYCKTWGQYWQV
amino acids 209-217 of gplOOP'117 GVRIVVEYCITDQVPFSV, ITDQVPFSVGVRIVVEYCITDQVPFSV
amino acids 280-288 of gplOOP'' GVRIVVEYCYLEPGPVTA, YLEPGPVTAGVRIVVEYCYLEPGPVTA
amino acids 457-466 of gp1001'117 GVRIVVEYCLLDGTATLRL, LLDGTATLRLGVRIVVEYCLLDGTATLRL
oe tµ.) tµ.) C35 Peptide/Epitope Exemplary Tumor Rejection Peptide Exemplary Polytopes amino acids 476-485 of gp100'17 GVRIVVEYCVLYRYGSFSV, VLYRYGSFSVGVRIVVEYCVLYRYGSFSV
amino acids 301-309 of PRAME . GVRIVVEYCLYVDSLFFL, LYVDSLFFLGVRIVVEYCLYVDSLFFL
amino acids 292-303 of MAGE-6 GVRIVVEYCKISGGPRISYPL, KISGGPRISYPLGVRIVVEYCKISGGPRISYPL
amino acids 157-167 of NY-ESO-1 GVRIVVEYCSLLMWITQCFL, SLLMWITQCFLGVRIVVEYCSLLMWITQCFL
amino acids 157-165 of NY-ESO-1 GVRIVVEYCSLLMWITQC, SLLMWITQCGVRIVVEYCSLLMWITQC

amino acids 155-163 of NY-ESO-1 GVRIVVEYCQLSLLMWIT, c7, QLSLLMWITGVRTVVEYCQLSLLM WIT
r;
t") amino acids 157-170 of NY-ESO-1 SLLMWITQCFLPVF

and amino acids 281-295 of MAGE-3 125 to C33 of SEQ ID NO:2 amino acids 161-169 of MAGE-1 IVVEYCEPCEADPTGHSY, 0 c7, IVVEYCEPC EADPTGHSYIVVEYCEPCEADPTGHSY
amino acids 230-238 of MAGE-1 IVVEYCEPCSAYGEPRKL, SAYGEPRKLIVVEYCEPCSAYGEPRKL
amino acids 168-176 of MAGE-3 IVVEYCEPCEVDPIGHLY, EVDPIGHLYIVVEYCEPCEVDPIGHLY
amino acids 271-279 of MAGE-3 IVVEYCEPCFLWGPRALV, FLWGPRALVIVVEYCEPCFLWGPRALV
amino acids 167-176 of MAGE-3 IVVEYCEPCMEVDPIGHLY, = MEVDPIGHLYIVVEYCEPCMEVDPIGHLY
oe C35 Peptide/Epitope Exemplary Tumor Rejection Peptide Exemplary Polytopes amino acids 2-10 of BAGE IVVEYCEPCAARAVFLAL, AARAVFLALIVVEYCEPCAARAVFLAL
amino acids 9-16 of GAGE-1,2 IVVEYCEPCYRPRPRRY, oe YRPRPRRYIVVEYCEPCYRPRPRRY
amino acids 11-20 of RAGE IVVEYCEPCSPSSNRIRNT, SPSSNRIRNTIVVEYCEPCSPS SNRIRNT
amino acids 23-32 of CDK4 IVVEYCEPCACDPHSGHFV, ACDPHSGHFVIVVEYCEPCACDPHSGHFV
amino acids 29-37 off3-catenin IVVEYCEPCSYLDSGIHF, SYLDSGIHFIVVEYCEPCSYLDSGIHF
amino acids 1-9 of tyrosinase IVVEYCEPC MLLAVLYCL, c7, MLLAVLYCLIVVEYCEPcMLLAVLYCL

amino acids 206-214 of tyrosinase IVVEYCEPCAFLPWHRLF, AFLPWHRLFIVVEYCEPCAFLPWHRLF
amino acids 56-70 of tyrosinase IVVEYCEPCQNILLSNAPLGPQFP, c7, QNILLSNAPLGPQFPIVVEYCEPCQNILLSNAPLGPQFP
aminb acids 448-462 of tyrosinase IVVEYCEPCDYSYLQDSDPDSFQD, DYSYLQDSDPDSFQDIVVEYCEPCDYSYLQDSDPDSFQD
amino acids 32-40 of Melan-A' IVVEYCEPCJLTVILGVL, JLTVILGVLIVVEYCEPCJLTVILGVL
amino acids 154-162 of gp100'17 IVVEYCEPCKTWGQYWQV, KTWGQYWQVIVVEYCEPCKTWGQYWQV
amino acids 209-217 of gp100P'117 IVVEYCEPCITDQVPFSV, ITDQVPFSVIVVEYCEPCITDQVPFSV
oe C35 Peptide/Epitope Exemplary Tumor Rejection Peptide Exemplary Polytopes amino acids 280-288 of gp100'117 IVVEYCEPCYLEPGPVTA, YLEPGPVTAIVVEYCEP CYLEPGPVTA
tµ.) amino acids 457-466 of gp100P'117 IVVEYCEPCLLDGTATLRL, LLD GTATLRLIVVEYCEP CLLDGTATLRL
amino acids 476-485 of gp100'17 IVVEYCEPCVLYRYGSFSV, VLYRYGSFSVIVVEYCEPCVLYRYGSFSV
amino acids 301-309 of PRAME IVVEYCEPCLYVD SLFFL, LYVD SLFFLIVVEYCEPCLYVD SLFFL
amino acids 292-303 of MAGE-6 IVVEYCEPCKISGGPRISYPL, KISGGPRISYPLIVVEYCEPCKISGGPRISYPL

co amino acids 157-167 of NY-ESO-1 IVVEYCEPCSLLMWITQCFL, co c7, co SLLMWITQCFLIVVEYCEPCSLLMWITQCFL
t\-) amino acids 157-165 of NY-ESO-1 IVVEYCEPCSLLMWITQC, SLLMWITQCIVVEYCEPCSLLMWITQC
HI
amino acids 155-163 of NY-ESO-1 IVVEYCEPCQLSLLMWIT, c7, QLSLLMWITIVVEYCEPCVQLSLLMWIT
amino acids 157-170 of NY-ESO-1 SLLMWITQCFLPVFIVVEYCEPCTSYVKVLHHMVKISG
and amino acids 281-295 of MAGE-3 T38-V46 of SEQ ID NO:2 amino acids 161-169 of MAGE-1 TYLELASAVEADPTGHSY, TYLELASAV EADPTGHSYTYLELASAVEADPTGHSY
amino acids 230-238 of MAGE-1 TYLELASAVSAYGEPRICL, SAYGEPRKLTYLELASAVSAYGEPRKL
amino acids 168-176 of MAGE-3 TYLELASAVEVDPIGHLY, EVDPIGHLYTYLELASAVEVDPIGHLY
oe tµ.) tµ.) C35 Peptide/Epitope Exemplary Tumor Rejection Peptide Exemplary Polytopes amino acids 271-279 of MAGE-3 TYLELASAVFLWGPRALV, FLWGPRALVTYLELASAVFLWGPRALV
tµ.) amino acids 167-176 of MAGE-3 TYLELASAVMEVDPIGHLY, oe MEVDPIGHLYTYLELASAVMEVDPIGHLY
amino acids 2-10 of BAGE TYLELASAVAARAVFLAL, AARAVFLALTYLELASAVAARAVFLAL
amino acids 9-16 of GAGE-1,2 TYLELASAVYRPRPRRY, YRPRPRRYTYLELASAVYRPRPRRY
amino acids 11-20 of RAGE TYLELASAVSPS SNRLRNT, SP S SNRIRNTTYLELASAVSPS SNRIRNT

co amino acids 23-32 of CDK4 TYLELASAVACDPHSGHFV, co c7, A CDPH S GHFVTYLELASAVACDPHS GHFV

amino acids 29-37 of (3-catenin TYLELASAVSYLD S GIEIF, SYLD S GIHFTYLELASAVSYLD S GIHF
amino acids 1-9 of tyrosinase TYLELASAVMLLAVLYCL, c7, MLLAVLYCLTYLELASAVMLLAVLYCL
amino acids 206-214 of tyrosinase TYLELASAVAFLPWHRLF, AFLPWHRLFTYLELASAVAFLPWHRLF
amino acids 56-70 of tyrosinase TYLELASAVQNILLSNAPLGPQFP, QNILLSNAPLGPQFPTYLELASAVQNILLSNAPLGPQFP
amino acids 448-462 of tyrosinase TYLELASAVDYSYLQDSDPDSFQD, DYSYLQD SDPD SFQDTYLELASAVDYSYLQDSDPDSFQD
amino acids 32-40 of Me1an-AmART-1 TYLELASAVJLTVILGVL, JLTVILGVLTYLELASAVJLTVILGVL
tµ.) tµ.) C35 Peptide/Epitope Exemplary Tumor Rejection Peptide Exemplary Polytopes amino acids 154-162 of gp1001'117 TYLELASAVKTWGQYWQV, KTWGQYWQVTYLELASAVKTWGQYWQV
amino acids 209-217 of gp100P'117 TYLELASAVITDQVPFSV, oe ITD QVPFSVTYLELASAVITD QVPFSV
amino acids 280-288 of gp100Pme 1 17 TYLELASAVYLEPGPVTA, YLEPGPVTATYLELASAVYLEPGPVTA
amino acids 457-466 of gp100'117 TYLELASAVLLDGTATLRL, LLD GTATLRLTYLELASAVLLDGTATLRL
amino acids 476-485 of gp 1 00Pme117 TYLELASAVVLYRYGSFSV, VLYRYGSFSVTYLELASAVVLYRYGSFSV

co amino acids 301-309 of PRAME TYLELASAVLYVD SLFFL, uco c7, LYVD SLFFLTYLELASAVLYVD SLFFL
ts.) amino acids 292-303 of MAGE-6 TYLELASAVKIS GGPRISYPL, KIS GGPRISYPLTYLELASAVKIS GGPRISYPL
amino acids 157-167 of NY-ESO-1 TYLELASAVSLLMWITQCFL, c7, SLLMWITQCFLTYLELASAVSLLMWITQCFL
amino acids 157-165 of NY-ESO-1 TYLELASAVSLLMWITQC, SLLMWITQCTYLELASAVSLLMWITQC
amino acids 155-163 of NY-ESO-1 TYLELASAVQLSLLMWIT, QLSLLMWITTYLELASAVQLSLLMWIT
amino acids 157-170 of NY-ESO-1 SLLMWITQCFLPVF
TYLELASAVTSYVKVLHEMVKISG

and amino acids 281-295 of MAGE-3 oe C35 Peptide/Epitope Exemplary Tumor Rejection Peptide Exemplary Polytopes G61-I69 of SEQ ID NO:2 amino acids 161-169 of MAGE-1 GTGAFEIEIEADPTGHSY, GTGAFEIEI EADPTGHSYGTGAFETHEADPTGHSY
amino acids 230-238 of MAGE-1 GTGAFEIEISAYGEPRKL, tµ.) oe SAYGEPRKLGTGAFEIEISAYGEPRKL
amino acids 168-176 of MAGE-3 GTGAFEIEIEVDPIGHLY, EVDPIGHLYGTGAFEIEIEVDPIGHLY
amino acids 271-279 of MAGE-3 GTGAFEIEIFLWGPRALV, FLWGPRALVGTGAFEIEIFLWGPRALV
amino acids 167-176 of MAGE-3 GTGAFEIEIMEVDPIGHLY, MEVDPIGHLYGTGAFEIEIMEVDPIGHLY

co amino acids 2-10 of BAGE GTGAFEIEIAARAVFLAL, co AARAVFLALGTGAFEIEIAARAVFLAL
r,..;
=
,õ
amino acids 9-16 of GAGE-1,2 GTGAFEIEIYRPRPRRY, YRPRPRRYGTGAFEIEIYRPRPRRY
amino acids 11-20 of RAGE GTGAFEIEISPSSNRIRNT, c7, SP SSNRIRNTGTGAFEIEISP SSNRIRNT
amino acids 23-32 of CDK4 GTGAFEIEIACDPHSGHFV, ACDPHSGHFVGTGAFEIEIACDPHSGHFV
amino acids 29-37 of P-catenin GTGAFEIEISYLDSGIEIF, SYLDSGIHFGTGAFEIEISYLDSGIHF
amino acids 1-9 of tyrosinase GTGAFEIEIMLLAVLYCL, MLLAVLYCLGTGAFEIEIMLLAVLYCL

amino acids 206-214 of tyrosinase GTGAFEIEIAFLPWHRLF, AFLPWHRLFGTGAFEIEIAFLPWHRLF
oe tµ.) tµ.) C35 Peptide/Epitope Exemplary Tumor Rejection Peptide Exemplary Polytopes amino acids 56-70 of tyrosinase GTGAFEIEIQNILLSNAPLGP QFP, QNILLSNAPLGP QFPGTGAFEIEIQNILLSNAPLGPQFP
tµ.) amino acids 448-462 of tyrosinase GTGAFEIEID YSYLQD SDPD SF
QD, DYSYLQD SDPD SF QD GTGAFEIEIDYSYLQD SDPD SFQD
amino acids 32-40 of Melan-AmAR't GTGAFEIEIJLTVILGVL, JLTVILGVLGTGAFEIEULTVILGVL
amino acids 154-162 of gp100P'117 GTGAFEIEIKTWGQYWQV, KTWGQYWQVGTGAFEMIKTWGQYWQV
amino acids 209-217 of gplOOP'117 GTGAFEIEIITDQVPFSV, =
ITDQVPFSVGTGAFEIEIITD QVPFSV

co amino acids 280-288 of gp100'w GTGAFEIEIYLEPGPVTA, co . YLEPGPVTAGTGAFEIEIYLEPGPVTA
amino acids 457-466 of gp1001'117 GTGAFEIEILLDGTATLRL, LLD GTATLRLGTGAFEIEILLDGTATLRL
amino acids 476-485 of gplOOPmem GTGAFEIEIVLYRYGSFSV, c7, VLYRYGSFSVGTGAFEIEIVLYRYGSFSV
amino acids 301-309 of PRAME GTGAFEIEILYVD SLFFL, LYVDSLFFLGTGAFEIEILYVDSLFFL
amino acids 292-303 of MAGE-6 GTGAFEIEIKIS GGPRISYPL, KISGGPRISYPLGTGAFEIEIKISGGPRISYPL
amino acids 157-167 of NY-ES 0-1 GTGAFEIEISLLMWITQCFL, SLLMWITQCFLGTGAFEIEISLLMWITQCFL
amino acids 157-165 of NY-ESO-1 GTGAFEIEISLLMWITQC, SLLMWITQCGTGAFEIEISLLMWITQC
oe tµ.) c.;11 tµ.) C35 Peptide/Epitope Exemplary Tumor Rejection Peptide Exemplary Polytopes amino acids 155-163 of NY-ESO-1 GTGAFEIEIQLSLLMWIT, QLSLLMWITGTGAFEIEIQLSLLMWIT
amino acids 157-170 of NY-ESO-1 SLLMWITQCFLPVFGTGAFEIEITSYVKVLHHMVKISG tµ.) oe and amino acids 281-295 of MAGE-3 F65-L73 of SEQ ID NO:2 amino acids 161-169 of MAGE-1 FEIEINGOLEADPTGHSY, FEIEINGQL EADPTGHSYFEIEINGOLEADPTGHSY
amino acids 230-238 of MAGE-1 FEIEINGOLSAYGEPRKL, = SAYGEPRKLFEIEINGOLSAYGEPRKL
amino acids 168-176 of MAGE-3 FEIEINGQLEVDPIGHLY, EVDPIGHLYFEIEINGOLEVDPIGHLY

co amino acids 271-279 of MAGE-3 FEIEINGOLFLWGPRALV, co FLWGPRALVFEIEINGQLFLWGPRALV
amino acids 167-176 of MAGE-3 FEIEINGQLMEVDPIGHLY, MEVDPIGHLYFErEINGQLMEVDPIGHLY
amino acids 2-10 of BAGE FEIEINGOLAARAVFLAL, c7, AARAVFLALFELEINGOLAARAVFLAL
amino acids 9-16 of GAGE-1,2 FErEINGQLYRPRPRRY, YRPRPRRYFEIEINGQLYRPRPRRY
amino acids 11-20 of RAGE FEIEINGOLSPSSNRIRNT, SPSSNRIRNTFEIEJNGOLSPSSNRIRNT
amino acids 23-32 of CDK4 FEIEINGQLACDPHSGHFV, ACDPHSGHFVFErEINGOLACDPHSGHFV

amino acids 29-37 off3-catenin FEIEINGQLSYLDSGIHF, S YLDSGIHFFEIEINGOLSYLDSGIFIF
oe JI
tµ.) tµ.) C35 Peptide/Epitope Exemplary Tumor Rejection Peptide Exemplary Polytopes amino acids 1-9 of tyrosinase FEIEINGOLMLLAVLYCL, MLLAVLYCLFEIEINGQLMLLAVLYCL
tµ.) amino acids 206-214 of tyrosinase FEIEINTGQLAFLPWHRLF, AFLPWHRLFFEIEINGQLAFLPWHRLF
amino acids 56-70 of tyrosinase FEIEINGQLQNILLSNAPLGPQFP, =
QNILLSNAPLGPQFPFEIEINGQLQNILLSNAPLGPQFP
amino acids 448-462 of tyrosinase FEIEINGQLDYSYLQDSDPDSFQD, DYSYLQDSDPDSFQDFEIEINGQLDYSYLQDSDPDSFQD
amino acids 32-40 of Melan-A' FEIEINGQLJLTVILGVL, JLTVILGVLFEIEINGQLJLTVILGVL
co amino acids 154-162 of gp100'17 FEIEINGQLKTWGQYWQV, co c7, co KTWGQYWQVFEIEINGQLKTWGQYWQV
? "
amino acids 209-217 of gp100' FEMINGQLITDQVPFSV, ITDQVPFSVFEIEINGOLITDQVPFSV
amino acids 280-288 of gplOOP'117 FEIEINGQLYLEPGPVTA, c7, YLEPGPVTAFEIEINGOLYLEPGPVTA
amino acids 457-466 of gp100'1'7 FEIEINGQLLLDGTATLRL, LLDGTATLRLFEIEINGQLLLDGTATLRL
amino acids 476-485 of gplOOPmem FEIEINGQLVLYRYGSFSV, VLYRYGSFSVFEIEINGQLVLYRYGSFSV
amino acids 301-309 of PRAME FELEINGOLLYVDSLFFL, LYVDSLFFLFEIEINGQLLYVDSLFFL
amino acids 292-303 of MAGE-6 FEIELNGQLKISGGPRISYPL, KISGGPRISYPLFEIEINGQLKISGGPRISYPL
oe tµ.) tµ.) C35 Peptide/Epitope Exemplary Tumor Rejection Peptide Exemplary Polytopes amino acids 157-167 of NY-ESO-1 FEIEINGOLSLLMWITQCFL, SLL1VfWITQCFLFEIEINGOLSLLMWITQCFL
tµ.) amino acids 157-165 of NY-ESO-1 FEIEINGOLSLLMWITQC, SLLMWITQCFEIEINGOLSLLMWITQC
amino acids 155-163 of NY-ESO-1 = FEIEINGOLQLSLLMWIT, QLSLLMWITFEIEINGQLQLSLLMWIT
amino acids 157-170 of NY-ESO-1 SLLMWITQCFLPVFFEIEINGOLTSYVKVLHHMVKISG
and amino acids 281-295 of MAGE-3 I67-F75 of SEQ ID NO:2 amino acids 161-169 of MAGE-1 IEINGOLVFEADPTGHSY, IEINGQLVF EADPTGHSYIEINGOLVFEADPTGHSY

co amino acids 230-238 of MAGE-1 T.EINGOLVFSAYGEPRKL, co =
SAYGEPRKLIEINGOLVFSAYGEPRKL
amino acids 168-176 of MAGE-3 IEINGOLVFEVDPIGHLY, EVDPIGHLYIEINGOLVFEVDPIGHLY
amino acids 271-279 of MAGE-3 IEINGOLVFFLWGPRALV, c7, FLWGPRALVIONGOLVFFLWGPRALV
amino acids 167-176 of MAGE-3 IEINGOLVFMEVDPIGHLY, MEVDPIGHLYIEINGOLVFMEVDPIGHLY
amino acids 2-10 of BAGE IEINGOLVFAARAVFLAL, AARAVFLALIEINGOLVFAARAVFLAL
amino acids 9-16 of GAGE-1,2 IEINGQLVFYRPRPRRY, YRPRPRRYIEINGQLVFYRPRPRRY
amino acids 11-20 of RAGE IEINGQLVF SP S SNRIRNT, SP S SNRIRNTIEING OLVF SP SSNRIRNT
oe tµ.) tµ.) C35 Peptide/Epitope Exemplary Tumor Rejection Peptide Exemplary Polytopes amino acids 23-32 of CDK4 IEINGQLVFCDPHSGHFV, ACDPHSGHFVIEINGQLVFACDPHSGHFV
tµ.) amino acids 29-37 of 13-catenin MINGOLVFSYLDSGIHF, SYLDSGIHFIEINGQLVFSYLDSGIHF
amino acids 1-9 of tyrosinase MINGQLVFMLLAVLYCL, MLLAVLYCLIEINGQLVFMLLAVLYCL
amino acids 206-214 of tyrosinase IEINGQLVFAFLPWHRLF, AFLPWHRLFIEINGOLVFAFLPWHRLF
amino acids 56-70 of tyrosinase IENGOLVFQNILLSNAPLGPQFP, QNILLSNAPLGPQFPIEINGQLVFQNILLSNAPLGPQFP
co amino acids 448-462 of tyrosinase MINGQLVFDYSYLQDSDPDSFQD, co c7, DYSYLQDSDPDSFQDIEINGOLVFDYSYLQDSDPDSFQD
u.) amino acids 32-40 of Melan-AmART-I IENGQLVFJLTVILGVL, JLTVILGVLIEINGOLVFJLTVILGVL
amino acids 154-162 of gp100'117 IEINGQLVFKTWGQYWQV, c7, KTWGQYWQVIETNGQLVFKTWGQYWQV
amino acids 209-217 of gp100Pme117 IE1NGQLVFITDQVPFSV, ITDQVPFSVIEINGQINFITDQVPFSV
amino acids 280-288 of gplOOPmel" IEINGOLVFYLEPGPVTA, YLEPGPVTAMINGQLVFYLEPGPVTA
amino acids 457-466 of gp100Pme117 IEINGOLVFLLDGTATLRL, LLDGTATLRLIEINGQLVFLLDGTATLRL
amino acids 476-485 of gp Memel 17 IEINGQLVFVLYRYGSFSV, VLYRYGSFSVIEINGQLVFVLYRYGSFSV
oe tµ.) tµ.) C35 Peptide/Epitope Exemplary Tumor Rejection Peptide Exemplary Polytopes amino acids 301-309 of PRAME IEINGOLVFLYVDSLFFL, LYVDSLFFLIEINGQLVFLYVDSLFFL
tµ.) amino acids 292-303 of MAGE-6 IEINGQLVFICISGGPRISYPL, oe KISGGPRISYPLIEINGQLVFKISGGPRISYPL
amino acids 157-167 of NY-ESO-1 IEINGQLVFSLLMWITQCFL, SLLMWITQCFLIEINGQLVFSLLMWITQCFL
amino acids 157-165 of NY-ESO-1 IETNGQLVFSLLMWITQC, SLLMWITQCIEINGQLVFSLLMWITQC
amino acids 155-163 of NY-ESO-1 IEINGQLVFQLSLLMWIT, QLSLLMWI l'IEINGQLVFQLSLLMWIT
co co amino acids 157-170 of NY-ESO-1 SLLMWITQCFLPVFIE1NGOLVFTSYVKVLHHMVKISG c7, co and amino acids 281-295 of MAGE-3 w K77-Y85 of SEQ ID NO:2 amino acids 161-169 of MAGE-1 KLENGGFPYEADPTGHSY, 0 KLENGGFPY EADPTGHSYKLENGGFPYEADPTGHSY
amino acids 230-238 of MAGE-1 ICLENGGFPYSAYGEPRKL, c7, SAYGEPRKLKLENGGFPYSAYGEPRKL
amino acids 168-176 of MAGE-3 KLENGGFPYEVDPIGHLY, EVDPIGHLYKLENGGFPYEVDPIGHLY
amino acids 271-279 of MAGE-3 ICLENGGFPYFLWGPRALV, FLWGPRALVKLENGGFPYFLWGPRALV
amino acids 167-176 of MAGE-3 KLENGGFPYMEVDPIGHLY, MEVDPIGHLYICLENGGFPYMEVDPIGHLY
amino acids 2-10 of BAGE KLENGGFPYAARAVFLAL, AARAVFLALKLENGGFPYAARAVFLAL
tµ.) tµ.) . .
_______________________________________________________________________________ ______________________________ C35 Peptide/Epitope Exemplary Tumor Rejection Peptide Exemplary Polytopes amino acids 9-16 of GAGE-1,2 KLENGGFPYYRPRPRRY, YRPRPRRYKLENGGFPYYRPRPRRY
tµ.) amino acids 11-20 of RAGE KLENGGFPYSPS SNRIRNT, oe SP S SNRIRNTKLENGGFPYSPSSNRIRNT
amino acids 23-32 of CDK4 KLENGGFPYACDPHS GHFV, ACDPHS GHFVKLENGGFPYACDPHSGHFV
amino acids 29-37 off3-catenin KLENGGFPYSYLD S G1HF, amino acids 1-9 of tyrosinase KLENGGFPYMLLAVLYCL, MLLAVLYCLKLENGGFPYMLLAVLYCL
co co amino acids 206-214 of tyrosinase KLENGGFPYAFLPWHRLF, c7, op AFLPWBRLFKLENGGFPYAFLPWHRLF
w 1\) amino acids 56-70 of tyrosinase KLENGGFPYQNILLSNAPLGPQFP, QNILLSNAPLGPQFPICLENGGFPYQNILLSNAPLGPQFP
amino acids 448-462 of tyrosinase KLENGGFPYDYSYLQDSDPDSFQD, c7, DYSYLQD SDPD SF QDKL,ENGGFPYDYSYLQD SDPDSFQD
amino acids 32-40 of Melan-AmAR' KLENGGFPYJLTVILGVL, JLTVILGVLKLENGGFPYJLTVILGVL
amino acids 154-162 of gp100'117 KLENGGFPYKTWGQYWQV, KTWGQYWQVKLENGGFPYKTWGQYWQV
amino acids 209-217 of gp100'17 KLENGGFPYIIDQVPFSV, ITDQVPFSVICLENGGFPYITDQVPFSV
amino acids 280-288 of gp100'117 KLENGGFPYYLEPGPVTA, YLEPGPVTAKLENGGFPYYLEPGPVTAJI
tµ.) C35 Peptide/Epitope Exemplary Tumor Rejection Peptide Exemplary Polytopes amino acids 457-466 of gp100'117 KLENGGFPYLLDGTATLRL, LLDGTATLRLKLENGGFPYLLDGTATLRL
tµ.) amino acids 476-485 of gplOOPmel 17 KLENGGFPYVLYRYGSFSV, oe VLYRYGSFSVKLENGGFPYVLYRYGSFSV
amino acids 301-309 of PRAME KLENGGFPYLYVDSLFFL, LYVDSLFFLICLENGGFPYLYVDSLFFL
amino acids 292-303 of MAGE-6 ICLENGGFPYKISGGPRISYPL, KISGGPRISYPLKLENGGFPYKISGGPRISYPL
amino acids 157-167 of NY-ESO-1 KLENGGFPYSLLMWITQCFL, SLLMWITQCFLKLENGGFPYSLLMWITQCFL

co amino acids 157-165 of NY-ESO-1 KLENGGFPYSLLMWITQC, co c7, op SLLMWITQCKLENGGFPYSLLMWITQC
1,3 amino acids 155-163 of NY-ESO-1 KLENGGFPYQLSLLMWIT, QLSLLMWITKLENGGFPYQLSLLMWIT
amino acids 157-170 of NY-ESO-1 SLLMWITQCFLPVF

c7, and amino acids 281-295 of MAGE-3 Q72-E86 of SEQ ID NO:2 amino acids 161-169 of MAGE-1 OLVFSKLENGGFPYEEADPTGHSY, QLVFSKLENGGFPYE
EADPTGHSYQLVFSKLENGGFPYEEADPTGHSY
amino acids 230-238 of MAGE-1 OLVFSICLENGGFPYESAYGEPRKL, SAYGEPRKLOLVFSKLENGGFPYESAYGEPRKL
amino acids 168-176 of MAGE-3 OLVFSICLENGGFPYEEVDPIGHLY, EVDPIGHLYOLVFSKLENGGFPYEEVDPIGHLY
amino acids 271-279 of MAGE-3 OLVFSICLENGGFPYEFLWGPRALV, FLWGPRALVOLVFSKLENGGFPYEFLWGPRALV
oe tµ.) tµ.) C35 Peptide/Epitope Exemplary Tumor Rejection Peptide Exemplary Polytopes amino acids 167-176 of MAGE-3 QLVFSKLENGGFPYEMEVDPIGHLY, MEVDPIGHLYQLVFSKLENGGFPYEMEVDPIGHLY
tµ.) amino acids 2-10 of BAGE QLVFSICLENGGFPYEAARAVFLAL, oe AARAVFLALQLVFSKLENGGFPYEAARAVFLAL
amino acids 9-16 of GAGE-1,2 QLVFSKLENGGFPYEYRPRPRRY, YRPRPRRYOLVFSKLENGGFPYEYRPRPRRY
amino acids 11-20 of RAGE QLVFSKLENGGFPYESPSSNRIRNT, SPSSNRIRNTQLVFSKLENGGFPYESPSSNRIRNT
amino acids 23-32 of CDK4 QLVFSKLENGGFPYEACDPHSGHFV, ACDPHSGHFVQLVFSICLENGGFPYEACDPHSGHFV
co co amino acids 29-37 off3-catenin QLVFSKLENGGFPYESYLDSGIHF, c7, co SYLDSGIHFOLVFSKLENGGFPYESYLDSGIHF
w amino acids 1-9 of tyrosinase QLVFSKLENGGFPYEMLLAVLYCL, MLLAVLYCLQLVFSKLENGGFPYEMLLAVLYCL
amino acids 206-214 of tyrosinase QLVFSKLENGGFPYEAFLPWHRLF, c7, AFLPWHRLFQLVFSICLENGGFPYEAFLPWHRLF
amino acids 56-70 of tyrosinase QLVFSKLENGGFPYEQNILLSNAPLGPQFP, QNILLSNAPLGPQFPQLVFSKLENGGFPYEQNILLSNAPLGPQFP
amino acids 448-462 of tyrosinase OLVFSKLENGGFPYEDYSYLQDSDPDSFQD, DYSYLQDSDPDSFQDQLVFSKLENGGFPYEDYSYLQDSDPDSFQD
amino acids 32-40 of Melan-A' QLVFSICLENGGFPYEJLTVILGVL, JLTVILGVLQLVFSKLENGGFPYEJLTVILGVL
amino acids 154-162 of gp100Pme117 QLVFSICLENGGFPYEKTWGQYWQV, KTWGQYWQVQLVFSKLENGGFPYEKTWGQYWQV
tµ.) tµ.) C35 Peptide/Epitope Exemplary Tumor Rejection Peptide Exemplary Polytopes amino acids 209-217 of gp100Pme' QLVFSKLENGGFPYEITDQVPFSV, ITDQVPFSVOLVESKLENGGFPYEITD QVPFSV
tµ.) amino acids 280-288 of gp100Pmem QLVFSKLENGGFPYEYLEPGPVTA, oe YLEPGPVTAQLVFSKLENGGFPYEYLEPGPVTA
amino acids 457-466 of gp100P'117 QLVFSKLENGGFPYELLDGTATLRL, LLDGTATLRLQLVFSKLENGGFPYELLDGTATLRL
amino acids 476-485 of gp100'1" QLVFSKLENGGFPYEVLYRYGSFSV, VLYRYGSFSVQLVFSKLENGGFPYEVLYRYGSFSV
amino acids 301-309 of PRAME OLVFSKLENGGFPYELYVD SLFFL, LYVD SLFFLQLVFSKLENGGFPYELYVDSLFFL

co co amino acids 292-303 of MAGE-6 QLVFSKLENGGFPYEKIS GGPRISYPL, c7, OD
KIS GGPRISYPLQLVFSKLENGGFPYEKIS GGPRISYPL

amino acids 157-167 of NY-ESO-1 QLVFSKLENGGFPYESLLMWITQCFL, SLLMWITQCFLQLVFSKLENGGFPYESLLMWITQCFL
amino acids 157-165 of NY-ESO-1 QLVFSICLENGGFPYESLLMWITQC, c7, SLLMWITQCQLVFSKLENGGFPYESLLMWITQC
amino acids 155-163 of NY-ESO-1 OLVFSKLENGGFPYEQLSLLMWIT, QLSLLMWITQLVFSKLENGGFPYEQLSLLMWIT
amino acids 157-170 of NY-ESO-1 SLLMWITQCFLPVFOLVFSKLENGGFPYETSYVKVLBEMVKISG
and amino acids 281-295 of MAGE-3 oe C35 Peptide/Epitope Exemplary Tumor Rejection Peptide Exemplary Polytopes G81-L89 of SEQ ID NO:2 amino acids 161-169 of MAGE-1 GGFPYEKDLEADPTGHSY, GGFPYEKDL EADPTGHSYGGFPYEKDLEADPTGHSY
amino acids 230-238 of MAGE-1 GGFPYEKDLSAYGEPRKL, oo SAYGEPRKLGGFPYEKDLSAYGEPRKL
amino acids 168-176 of MAGE-3 GGFPYEKDLEVDPIGHLY, EVDPIGHLYGGFPYEKDLEVDPIGHLY
amino acids 271-279 of MAGE-3 GGFPYEKDLFLWGPRALV, FLWGPRALVGGFPYEKDLFLWGPRALV
amino acids 167-176 of MAGE-3 GGFPYEKDLMEVDPIGHLY, MEVDPIGHLYGGFPYEKDLMEVDPIGHLY
amino acids 2-10 of BAGE GGFPYEKDLAARAVFLAL, co AARAVFLALGGFPYEKDLAARAVFLAL
w oo amino acids 9-16 of GAGE-1,2 GGFPYEKDLYRPRPRRY, YRPRPRRYGGFPYEKDLYRPRPRRY
amino acids 11-20 of RAGE GGFPYEKDLSPSSNRIRNT, SPSSNRIRNTGGFPYEKDLSPSSNRIRNT
amino acids 23-32 of CDK4 GGFPYEKDLACDPHSGHFV, ACDPHSGHFVGGFPYEKDLACDPHSGHFV
amino acids 29-37 of P-catenin GGFPYEICDLSYLDSGIEIF, SYLDSGIHFGGFPYEKDLSYLDSGIHF
amino acids 1-9 of tyrosinase GGFPYEKDLMLLAVLYCL, MLLAVLYCLGGFPYEKDLMLLAVLYCL
amino acids 206-214 of tyrosinase GGFPYEKDLAFLPWHRLF, AFLYVVHRLFGGFPYEKDLAFLPWHRLF
oo C35 Peptide/Epitope Exemplary Tumor Rejection Peptide Exemplary Polytopes amino acids 56-70 of tyrosinase GGFPYEKDLQNILLSNAPLGPQFP, QNILLSNAPLGPQFPGGFPYEKDLQNILLSNAPLGPQFP
amino acids 448-462 of tyrosinase GGFPYEKDLDYSYLQDSDPDSFQD, tµ.) DYSYLQDSDPDSFQDGGFPYEKDLDYSYLQDSDPDSFQD
amino acids 32-40 of Me1an-AmART4 GGFPYEKDLJLTVILGVL, JLTVILGVLGGFPYEICDULTVILGVL
amino acids 154-162 of gp100Pmell7 GGFPYEICDLKTWGQYWQV, KTWGQYWQVGGFPYEKDLKTWGQYWQV
amino acids 209-217 of gp100'17 GGFPYE1CDLITDQVPFSV, ITDQVPFSVGGFPYEKDLITDQ VPFSV

co amino acids 280-288 of gp100''' GGFPYEKDLYLEPGPVTA, I CO
(3) YLEPGPVTAGGFPYEKDLYLEPGPVTA
Lo co "
"
amino acids 457-466 of gp100P'117 GGFPYEKDLLLDGTATLRL, LLDGTATLRLGGFPYEKDLLLDGTATLRL
amino acids 476-485 of gplooPme117 GGFPYEKDLVLYRYGSFSV, VLYRYGSFSVGGFPYEKDLVLYRYGSFSV
amino acids 301-309 of PRAME GGFPYEKDLLYVDSLFFL, LYVDSLFFLGGFPYEKDLLYVDSLFFL
amino acids 292-303 of MAGE-6 GGFPYEKDLKISGGPRISYPL, KISGGPRISYPLGGFPYEKDLKISGGPRISYPL
amino acids 157-167 of NY-ESO-1 GGFPYEKDLSLLMWITQCFL, amino acids 157-165 of NY-ESO-1 GGFPYEKDLSLLMWITQC, SLLMWITQCGGFPYEKDLSLLMWITQC
oe tµ.) tµ.) C35 Peptide/Epitope Exemplary Tumor Rejection Peptide Exemplary Polytopes amino acids 155-163 of NY-ES 0-1 GGFPYEKDLQLSLLMWIT, QLSLLMWITGGFPYEKDLQLSLLMWIT
tµ.) amino acids 157-170 of NY-ES 0-1 SLLMWITQCFLPVFGGFPYEKDLTSYVKVLHHMVKIS G oe and amino acids 281-295 of MAGE-3 K104-C112 of SEQ ID NO:2 amino acids 161-169 of MAGE-1 KITNSRPP CEADPTGHSY, KITNSRPPC EADPTGHSYKITNSRPP CEADPTGHSY
amino acids 230-238 of MAGE-1 KITNSRPPCSAYGEPRICL, SAYGEPRKLKITNSRPPCSAYGEPRKL
amino acids 168-176 of MAGE-3 KITNSRPPCEVDPIGHLY, EVDPIGHLYKITNSRPPCEVDPIGHLY
co co amino acids 271-279 of MAGE-3 KITNSRPPCFLWGPRALV, c7, FLWGPRALVKITNSRPPCFLWGPRALV
1-11:

amino acids 167-176 of MAGE-3 KITNSRPPCMEVDPIGHLY, MEVDPIGHLYKITNSRPPCMEVDPIGHLY
amino acids 2-10 of BAGE KITNSRPPCAARAVFLAL, c7, AARAVFLALKITNSRPPCAARAVFLAL
amino acids 9-16 of GAGE-1,2 KITNSRPPCYRPRPRRY, YRPRPRRYKITNSRPPCYRPRPRRY
amino acids 11-20 of RAGE KITNSRPP C SP S SNRIRNT, SP S SNRIRNTKITNSRPPCSPS SNRIRNT
amino acids 23-32 of CDK4 KITNSRPPCACDPHS GHFV, ACDPHS GHFVKITNSRPPCACDPHS GHFV
amino acids 29-37 of f3-catenin KITNSRPPCSYLD S GIHF, SYLD S GIIIFKITNSRPP CSYLD SGIFIF
tµ.) tµ.) C35 Peptide/Epitope Exemplary Tumor Rejection Peptide Exemplary Polytopes amino acids 1-9 of tyrosinase KITNSRPPCMLLAVLYCL, MLLAVLYCLKITNSRPPCMLLAVLYCL
amino acids 206-214 of tyrosinase KITNSRPPCAFLPWHRLF, tµ.) oe AFLPWHRLFKITNSRPPCAFLPWHRLF
amino acids 56-70 of tyrosinase ICITNSRPPCQNILLSNAPLGPQFP, QNILLSNAPLGPQFPKITNSRPPCQNILLSNAPLGPQFP
amino acids 448-462 of tyrosinase KITNSRPPCDYSYLQDSDPDSFQD, DYSYLQDSDPDSFQDKITNSRPPCDYSYLQDSDPDSFQD
amino acids 32-40 of Melan-AmART-1 KITNSRPPCJLTVILGVL, JLTVILGVLKITNSRPPCJLTVILGVL

amino acids 154-162 of gpl00Pme117 KITNSRPPCKTWGQYWQV, co co KTWGQYWQVKITNSRPPCKTWGQYWQV
N.) amino acids 209-217 of gp100'" KITNSRPPCITDQVPFSV, ITDQVPFSVKITNSRPPCITDQVPFSV
amino acids 280-288 of gp100'117 KITNSRPPCYLEPGPVTA, YLEPGPVTAKITNSRPPCYLEPGPVTA
c7, amino acids 457-466 of gp100'1" KITNSRPPCLLDGTATLRL, LLDGTATLRLKITNSRPPCLLDGTATLRL
amino acids 476-485 of gp 1 00'1" KITNSRPPCVLYRYGSFSV, VLYRYGSFSVKITNSRPPCVLYRYGSFSV
amino acids 301-309 of PRAME KITNSRPPCLYVDSLFFL, LYVDSLFFLICITNSRPPCLYVDSLFFL

amino acids 292-303 of MAGE-6 KITNSRPPCKISGGPRISYPL, KISGGPRISYPLKITNSRPPCKISGGPRISYPL
oe tµ.) tµ.) C35 Peptide/Epitope Exemplary Tumor Rejection Peptide Exemplary Polytopes amino acids 157-167 of NY-ESO-1 KITNSRPPCSLLMWITQCFL, SLLMWITQCFLKITNSRPPCSLLMWITQCFL
amino acids 157-165 of NY-ESO-1 KITNSRPPCSLLMWITQC, tµ.) oe SLLMVVITQCKITNSRPPCSLLMWITQC
amino acids 155-163 of NY-ESO-1 KITNSRPPCQLSLLMWIT, QLSLLMWITKITNSRPPCQLSLLMWIT
amino acids 157-170 of NY-ESO-1 SLLMWITQCFLPVFKITNSRPPCTSYVKVLHHMVKISG
and amino acids 281-295 of MAGE-3 K104-V113 of SEQ ID NO:2 amino acids 161-169 of MAGE-1 KITNSRPPCVEADPTGHSY, KITNSRPPCV EADPTGHSYKITNSRPPCVEADPTGHSY

amino acids 230-238 of MAGE-1 KITNSRPPCVSAYGEPRKL, SAYGEPRKLKITNSRPPCVSAYGEPRKL
"
amino acids 168-176 of MAGE-3 KITNSRPPCVEVDPIGHLY, EVDPIGHLYKITNSRPPCVEVDPIGHLY
amino acids 271-279 of MAGE-3 KITNSRPPCVFLWGPRALV, FLWGPRALVKITNSRPPCVFLWGPRALV
amino acids 167-176 of MAGE-3 KITNSRPPCVMEVDPIGHLY, MEVDPIGHLYKITNSRPPCVMEVDPIGHLY
amino acids 2-10 of BAGE KITNSRPPCVAARAVFLAL, AARAVFLALKITNSRPPCVAARAVFLAL
amino acids 9-16 of GAGE-1,2 KITNSRPPCVYRPRPRRY, YRPRPRRYKITNSRPPCVYRPRPRRY

amino acids 11-20 of RAGE KITNSRPPCVSPSSNRIRNT, SPSSNRIRNTKTTNSRPPCVSPSSNRIRNT
oe tµ.) tµ.) C35 Peptide/Epitope Exemplary Tumor Rejection Peptide Exemplary Polytopes amino acids 23-32 of CDK4 KITNSRPPCVACDPHSGHFV, ACDPHSGHFVKITNSRPPCVACDPHSGHFV
amino acids 29-37 of P-catenin KITNSRPPCVSYLDSGIHF, c'e SYLDSGIHFKITNSRPPCVSYLDSGIHF
amino acids 1-9 of tyrosinase KITNSRPPCVMLLAVLYCL, MLLAVLYCLKITNSRPPCVMLLAVLYCL
amino acids 206-214 of tyrosinase KITNSRPPCVAFLPWHRLF, AFLPWHRLFKITNSRPPCVAFLPWHRLF
amino acids 56-70 of tyrosinase KITNSRPPCVQNILLSNAPLGPQFP, QNILLSNAPLGPQFPKITNSRPPCVQNILLSNAPLGPQFP

co amino acids 448-462 of tyrosinase KITNSRPPCVDYSYLQDSDPDSFQD, co c7, DYSYLQDSDPDSFQDKITNSRPPCVDYSYLQDSDPDSFQD

amino acids 32-40 of Me1an-AmART4 KITNSRPPCVJLTVILGVL, JLTVILGVLKITNSRPPCVJLTVILGVL
amino acids 154-162 of gp100'117 KITNSRPPCVKTWGQYWQV, c7, KTWGQYWQVKITNSRPPCVKTWGQYWQV
amino acids 209-217 of gp1001'117 KITNSRPPCVITDQVPFSV, ITDQVPFSVKITNSRPPCVITDQVPFSV
amino acids 280-288 of gp10OP'117 KITNSRPPCVYLEPGPVTA, YLEPGPVTAKITNSRPPCVYLEPGPVTA
amino acids 457-466 of gp100'117 KITNSRPPCVLLDGTATLRL, LLDGTATLRLKITNSRPPCVLLDGTATLRL
amino acids 476-485 of gp100'117 KITNSRPPCVVLYRYGSFSV, VLYRYGSFSVKITNSRPPCVVLYRYGSFSV
oe C35 Peptide/Epitope Exemplary Tumor Rejection Peptide Exemplary Polytopes amino acids 301-309 of PRAME KITNSRPPCVLYVDSLFFL, LYVDSLFFLKITNSRPPCVLYVDSLFFL
tµ.) amino acids 292-303 of MAGE-6 KITNSRPPCVKISGGPRISYPL, oe KISGGPRISYPLICITNSRPPCVKISGGPRISYPL
amino acids 157-167 of NY-ESO-1 KITNSRPPCVSLLMWITQCFL, SLLMWITQCFLICITNSRPPCVSLLMWITQCFL
amino acids 157-165 of NY-ESO-1 K1TNSRPPCVSLLMWITQC, SLLMWITQCKITNSRPPCVSLLMWITQC
amino acids 155-163 of NY-ESO-1 KITNSRPPCVQLSLLMWIT, QLSLLMWITKITNSRPPCVQLSLLMWIT
co co amino acids 157-170 of NY-ESO-1 SLLMWITQCFLPVFKITNSRPPCVTSYVKVLHHMVKISG c7, and amino acids 281-295 of MAGE-3 1105- V113 of SEQ ID NO:2 amino acids 161-169 of MAGE-1 ITNSRPPCVEADPTGHSY, 0 ITNSRPPCV EADPTGHSYITNSRPPCVEADPTGHSY
amino acids 230-238 of MAGE-1 ITNSRPPCVSAYGEPRKL, c7, SAYGEPRKLITNSRPPCVSAYGEPRKL
amino acids 168-176 of MAGE-3 ITNSRPPCVEVDPIGHLY, EVDPIGHLYITNSRPPCVEVDPIGHLY
amino acids 271-279 of MAGE-3 ITNSRPPCVFLWGPRALV, FLWGPRALVITNSRPPCVFLWGPRALV
amino acids 167-176 of MAGE-3 ITNSRPPCVMEVDPIGHLY, MEVDPIGHLYITNSRPPCVMEVDPIGHLY
amino acids 2-10 of BAGE ITNSRPPCVAARAVFLAL, AARAVFLALITNSRPPCVAARAVFLAL
tµ.) tµ.) C35 Peptide/Epitope Exemplary Tumor Rejection Peptide Exemplary Polytopes 0 amino acids 9-16 of GAGE-1,2 ITNSRPPCVYRPRPRRY, YRPRPRRYITNSRPPCVYRPRPRRY
tµ.) oe amino acids 11-20 of RAGE ITNSRPPCVSPSSNRIRNT, SPSSNRIRNTITNSRPPCVSPSSNRIRNT
amino acids 23-32 of CDK4 ITNSRPPCVACDPHSGHFV, ACDPHSGHFVITNSRPPCVACDPHSGHFV
amino acids 29-37 of 13-catenin ITNSRPPCVSYLDSGIHF, SYLDSGIHFITNSRPPCVSYLDSGIHF
amino acids 1-9 of tyrosinase ITNSRPPCVMLLAVLYCL, MLLAVLYCLITNSRPPCVMLLAVLYCL
co co amino acids 206-214 of tyrosinase ITNSRPPCVAFLPWHRLF, c7, I
a) AFLPWHRLFITNSRPPCVAFLPWHRLF
"
1..) amino acids 56-70 of tyrosinase ITNSRPPCVQNILLSNAPLGPQFP, QNILLSNAPLGPQFPITNSRPPCVQNILLSNAPLGPQFP

amino acids 448-462 of tyrosinase ITNSRPPCVDYSYLQDSDPDSFQD, c7, DYSYLQDSDPDSFQDITNSRPPCVDSYLQDSDPDSFQD
amino acids 32-40 of Me1an-AmART4 ITNSRPPCVJLTVILGVL, JLTVILGVLITNSRPPCVJLTVILGVL
amino acids 154-162 of gp100' 17 ITNSRPPCVKTWGQYWQV, KTWGQYWQVITNSRPPCVKTWGQYWQV
amino acids 209-217 of gp100Pmen7 ITNSRPPCVITDQVPFSV, ITDQVPFSVITNSRPPCVITDQVPFSV
amino acids 280-288 of gpl 00Pmeh17 ITNSRPPCVYLEPGPVTA, YLEPGPVTAITNSRPPCVYLEPGPVTAJI
oe tµ.) tµ.) C35 Peptide/Epitope Exemplary Tumor Rejection Peptide Exemplary Polytopes amino acids 457-466 of gp100' ITNSRPPCVLLDGTATLRL, LLD GTATLRLITNSRPP CVLLDGTATLRL
tµ.) amino acids 476-485 of gplOOP'117 UNSRPPCVVLYRYGSFSV, VLYRYGSFSVITNSRPPCVVLYRYGSFSV
amino acids 301-309 of PRAME ITNSRPPCVLYVD SLFFL, LYVDSLFFLITNSRPPCVLYVD SLFFL
amino acids 292-303 of MAGE-6 ITNSRPPCVKIS GGPRISYPL, KIS GGPRISYPLITNSRPPCVKIS GGPRISYPL
amino acids 157-167 of NY-ESO-1 ITNSRPPCVSLLMWITQCFL, SLLMWITQCFLITNSRPPCVSLLMWITQCFL

co amino acids 157-165 of NY-ESO-1 ITNSRPPCVSLLMWITQC, co c7, SLLMWITQCITNSRPPCVSLLMWITQC
c?`
amino acids 155-163 of NY-ESO-1 ITNSRPPCVQLSLLMWIT, QLSLLMWITITNSRPPCVQLSLLMWIT
amino acids 157-170 of NY-ESO-1 c7, and amino acids 281-295 of MAGE-3 T101 -V113 of SEQ ID NO:2 amino acids 161-169 of MAGE,1 TLEKITNSRPP
CVEADPTGH SY, TLEKITNSRPP CV
EADPTGHSYTLEKITNSRPPCVEADPTGHSY
amino acids 230-238 of MAGE-1 TLEKITNSRPPCVSAYGEPRKL, SAYGEPRKLTLEKITNSRPPCVSAYGEPRKL
amino acids 168-176 of MA GE-3 TLEKITNSRPPCVEVDPIGHLY, EVDPIGHLY TLEKITNSRPPCVEVDPIGHLY
amino acids 271-279 of MAGE-3 TLEKITNSRPP CVFLWGPRALV, FLWGPRALVTLEKITNSRPPCVFLWGPRALV
oe tµ.) tµ.) C35 Peptide/Epitope Exemplary Tumor Rejection Peptide Exemplary Polytopes amino acids 167-176 of MAGE-3 TLEKITNSRPPCVMEVDPIGHLY, MEVDPIGHLYTLEKITNSRPPCVME VDPIGHLY
amino acids 2-10 of BAGE TLEKITNSRPPCVAARAVFLAL, oe AARAVFLALTLEKITNSRPPCVAARAVFLAL
amino acids 9-16 of GAGE-1,2 TLEICITNSRPPCVYRPRPRRY, amino acids 11-20 of RAGE TLEKITNSRPPCVSP S SNRIRNT, SP S SNRIRNTTLEKITNSRPP CVSPS SNRIRNT
amino acids 23-32 of CDK4 TLEKITNSRPPCVACDPHS GHFV, ACDPHSGHFVTLEKITNSRPPCVACDPHSGHFV

co amino acids 29-37 of13-catenin TLEICITNSRPPCVSYLD S GIHF, uco SYLD S GIHFTLEICITNSRPPCVSYLD S GIFT

"
amino acids 1-9 of tyrosinase TLEKITNSRPPCVMLLAVLYCL, MLLAVLYCLTLEKITNSRPP CVMLLAVLYCL
amino acids 206-214 of tyrosinase TLEKITNSRPPCVAFLPWHRLF, AFLPWHRLFTLEKITNSRPPCVAFLPWHRLF
c7, amino acids 56-70 of tyrosinase TLEKITNSRPPCVQNILLSNAPLGPQFP, QNILLSNAPLGPQFPTLEKITNSRPPCVQNILLSNAPLGPQFP
amino acids 448-462 of tyrosinase TLEKITNSRPP CVDYSYLQD SDPD SFQD, DYSYLQD SDPD SFQDTLEKITNSRPPCVDYSYLQDSDPD SFQD
amino acids 32-40 of Me1an-AmART4 TLEKITNSRPPCVJLTVILGVL, JLTVILGVLTLEKITNSRPPCVJLTVILGVL

amino acids 154-162 of gplOOPmeui7 TLEICITNSRPP CVKTWGQYWQV, KTWGQYWQVTLEKITNSRPPCVKTWGQYWQV
oe C35 Peptide/Epitope Exemplary Tumor Rejection Peptide Exemplary Polytopes amino acids 209-217 of gp100Pme117 TLEKITNSRPPCVITDQVPFSV, ITDQVPFSVTLEKITNSRPPCVITDQVPFSV
tµ.) amino acids 280-288 of gplOOPmen7 TLEKITNSRPPCVYLEPGPVTA, YLEPGPVTATLEKITNSRPPCVYLEPGPVTA
amino acids 457-466 of gplOOPniew TLEKITNSRPPCVLLDGTATLRL, LLDGTATLRLTLEKITNSRPPCVLLDGTATLRL
amino acids 476-485 of gp100'117 TLEKITNSRPPCVVLYRYGSFSV, VLYRYGSFSVTLEKITNSRPPCVVLYRYGSFSV
amino acids 301-309 of PRAME TLEKITNSRPPCVLYVDSLFFL, LYVDSLFFLTLEKITNSRPPCVLYVDSLFFL

co amino acids 292-303 of MAGE-6 TLEKITNSRPPCVKISGGPRISYPL, co KISGGPRISYPLTLEKITNSRPPCVKISGGPRISYPL

amino acids 157-167 of NY-ES0-1 TLEKITNSRPPCVSLLMWITQCFL, SLLMWITQCFLTLEKITNSRPPCVSLLMWITQCFL
amino acids 157-165 of NY-ESO-1 TLEKITNSRPPCVSLLMWITQC, c7, SLLMWITQCTLEKITNSRPPCVSLLMWITQC
amino acids 155-163 of NY-ESO-1 TLEKITNSRPPCVQLSLLMWIT, QLSLLMWITTLEKITNSRPPCVQLSLLMWIT
amino acids 157-170 of NY-ESO-1 SLLMWITQCFLPVF
TLEKITNSRPPCVTSYVKVLHHMVKISG
and amino acids 281-295 of MAGE-3 oe C35 Peptide/Epitope Exemplary Tumor Rejection Peptide Exemplary Polytopes 193 - V113 of SEQ ID NO:2 amino acids 161-169 of MAGE-1 1RRASNGETLEKITNSRPPCVEADPTGHSY, IRRASNGETLEKITNSRPP
EADPTGHSYIRRASNGETLEKITNSRPPCVEADPTGHSY
CV
tµ.) oe amino acids 230-238 of MAGE-1 IRRASNGETLEKITNSRPPCVSAYGEPRKL, SAYGEPRKLIRRASNGETLEKITNSRPPCVSAYGEPRKL
amino acids 168-176 of MAGE-3 IRRASNGETLEMNSRPPCVEVDPIGHLY, EVDPIGHLYIRRASNGETLEKITNSRPPCVEVDPIGHLY
amino acids 271-279 of MAGE-3 1RRASNGETLEKITNSRPPCVFLWGPRALV, FLWGPRALVIRRASNGETLEKITNSRPPCVFLWGPRALV
amino acids 167-176 of MAGE-3 IRRASNGETLEKITNSRPPCVMEVDPIGHLY, MEVDPIGHLYIRRASNGETLEKITNSRPPCVMEVDPIGHLY
co co amino acids 2-10 of BAGE
IRRASNGETLEKITNSRPPCVAARAVFLAL, c7, co AARAVFLALIRRASNGETLEKITNSRPPCVAARAVFLAL
"
%.0 1\.) amino acids 9-16 of GAGE-1,2 IRRASNGETLEKITNSRPPCVYRPRPRRY, YRPRPRRYIRRASNGETLEKITNSRPPCVYRPRPRRY
amino acids 11-20 of RAGE
IRRASNGETLEKITNSRPPCVSPSSNRIRNT, 0 c7, SP SSNRIRNTIRRASNGETLEKITNSRPPCVSPSSNRIRNT
amino acids 23-32 of CDK4 IRRASNGETLEKITNSRPPCVARDPHSGHFV, ARDPHSGHEVIRRASNGETLEKITNSRPPCVARDPHSGHEV
amino acids 29-37 of P-catenin IRRASNGETLEICITNSRPPCVSYLDSGMS, SYLDSGIHSIRRASNGETLEKITNSRPPCVSYLDSGIHS
amino acids 1-9 of tyrosinase IRRASNGETLEKITNSRPPCVMLLAVLYCL, MLLAVLYCLIRRASNGETLEKITNSRPPCVMLLAVLYCL
amino acids 206-214 of tyrosinase IRRASNGETLEKITNSRPPCVAFLPWHRLF, AFLPWHRLFIRRASNGETLEICITNSRPPCVAFLPWHRLFJI
oe tµ.) tµ.) C35 Peptide/Epitope Exemplary Tumor Rejection Peptide Exemplary Polytopes amino acids 56-70 of tyrosinase IRRASNGETLEKITNSRPPCVQNILLSNAPLGPQFP, QNILLSNAPLGPQFPIRRASNGETLEICITNSRPPCVQNILLSNAPLGPQFP
tµ.) amino acids 448-462 of tyrosinase IRRASNGETLEKITNSRPPCVDYSYLQDSDPDSFQD, oe DYSYLQDSDPDSFQDIRRASNGETLEKIFNSRPPCVDYSYLQDSDPDSFQD
amino acids 32-40 of Me1an-AmART-1 IRRASNGETLEKITNSRPPCVJLTVILGVL, JLTVILGVLIRRASNGETLEKITNSRPPCVJLTVILGVL
amino acids 154-162 of gp1001'117 IRRASNGETLEKITNSRPPCVKTWGQYWQV, KTWGQYWQVIRRASNGETLEKITNSRPPCVKTWGQYWQV
amino acids 209-217 of gp100'117 IRRASNGETLEKITNSRPPCVILITDQVPFSV, co amino acids 280-288 of gp1001'17 IRRASNGETLEKITNSRPPCVYLEPGPVTA, co c7, op YLEPGPVTAIRRASNGETLEKITNSRPPCVYLEPGPVTA

amino acids 457-466 of gplOOP'117 IRRASNGETLEKITNSRPPCVLLDGTATLRL, LLDGTATLRLIRRASNGETLEKITNSRPPCVLLDGTATLRL
amino acids 476-485 of gplOOPine' IRRASNGETLEKITNSRPPCVVLYRYGSFSV, c7, VLYRYGSFSVIRRASNGETLEKITNSRPPCVVLYRYGSFSV
amino acids 301-309 of PRAME IRRASNGETLEKITNSRPPCVLYVDSLFFL, LYVDSLFFLIRRASNGETLEKITNSRPPCVLYVDSLFFL
amino acids 292-303 of MAGE-6 IRRASNGETLEKITNSRPPCVICISGGPRISYPL, ICISGGPRISYPLIRRASNGETLEKITNSRPPCVICISGGPRISYPL
amino acids 157-167 of NY-ESO-1 IRRASNGETLEKITNSRPPCVSLLMWITQCFL, SLLMWITQCFLIRRASNGETLEKITNSRPPCVSLLMWITQCFL
amino acids 157-165 of NY-ESO-1 IRRASNGETLEKITNSRPPCVSLLMWITQC, SLLMWITQCIRRASNGETLEICITNSRPPCVSLLMWITQC
tµ.) tµ.) C35 Peptide/Epitope Exemplary Tumor Rejection Peptide Exemplary Polytopes amino acids 155-163 of NY-ESO-1 IRRASNGETLEKITNSRPPCVQLSLLMWIT, QLSLLMWITIRRASNGETLEKITNSRPPCVQLSLLMWIT
amino acids 157-170 of NY-ESO-1 SLLMWITQCFLPVFIRRASNGETLEKITNSRPPCVTSYVKVLHHMVKISG tµ.) oe and amino acids 281-295 of MAGE-3 D88 - V113 of SEQ ID NO:2 amino acids 161-169 of MAGE-1 DLIEAIRRASNGETLEKITNSRPPCVEADPTGHSY, DLIEAIRRASNGETLEKIT
EADPTGHSYDLIEAIRRASNGETLEICIINSRPPCVEADPTGHSY
NSRPPCV
amino acids 230-238 of MAGE-1 DLIEAIRRASNGETLEKITNSRPPCVSAYGEPRKL, SAYGEPRKLDLIEAIRRASNGETLEKITNSRPPCVSAYGEPRICL
amino acids 168-176 of MAGE-3 DLIEAIRRASNGETLEKITNSRPPCVEVDPIGHLY, EVDPIGHLYDLIEMRRASNGETLEKITNSRPPCVEVDPIGHLY
amino acids 271-279 of MAGE-3 DLIEAIRRASNGETLEKITNSRPPCVFLWGPRALV, co co (31 FLWGPRALVDLIEAIRRASNGETLEKITNSRPPCVFLWGPRALV
op amino acids 167-176 of MAGE-3 DLIEALRRASNGETLEKITNSRPPCVMEVDPIGHLY, MEVDPIGHLYDLIEAIRRASNGETLEKITNSRPPCVMEVDPIGHLY
amino acids 2-10 of BAGE
DLIEAIRRASNGETLEICITNSRPPCVAARAVFLAL, AARAVFLALDLIEAIRRASNGETLEKITNSRPPCVAARAVFLAL
amino acids 9-16 of GAGE-1,2 DLIEAIRRASNGETLEKITNSRPPCVYRPRPRRY, YRPRPRRYDLIEAIR_RASNGETLEKITNSRPPCVYRPRPRRY
amino acids 11-20 of RAGE
DLIEAIRRASNGETLEICITNSRPPCVSPSSNRIRNT, SP SSNRIRNTDLIEAIRRASNGETLEICITNSRPPCVSP SSNRIRNT
amino acids 23-32 of CDK4 DLIEAIRRASNGETLEKITNSRPPCVARDPHSGHFV, ARDPHSGHFVDLIEAIRRASNGETLEKIINSRPPCVARDPHSGHFV

amino acids 29-37 off3-catenin DLIEAIRRASNGETLEKITNSRPPCVSYLDSGIHS, SYLDSGIEISDLIEAIRRASNGETLEKITNSRPPCVSYLDSGIHS
oe tµ.) tµ.) C35 Peptide/Epitope Exemplary Tumor Rejection Peptide Exemplary Polytopes amino acids 1-9 of tyrosinase DLIEAIRRASNGETLEKITNSRPPCVMLLAVLYCL, MLLAVLYCLDLIEAIRRASNGETLEKITNSRPPCVMLLAVLYCL
tµ.) amino acids 206-214 of tyrosinase DLIEAIRRASNGETLEKITNSRPPCVAFLPWHRLF, oe AFLPWHRLFDLIEAIRRASNGETLEKITNSRPPCVAFLPWHRLF
amino acids 56-70 of tyrosinase DLIEAIRRASNGETLEICITNSRPPCVQNILLSNAPLGPQFP, QNILLSNAPLGP QFPDLLEAIRRASNGETLEKITNSRPP CVQNILLSNAPLGP QFP
amino acids 448-462 of tyrosinase DLIEAIRRASNGETLEKITNSRPPCVDYSYLQDSDPDSFQD, DYSYLQD SDPD SFQDDLIEAIRRASNGETLEKITNSRPPCVDYSYLQD SDPDSFQD
amino acids 32-40 of Me1an-AmART1 DLIEAIRRASNGETLEKITNSRPPCVJLTVILGVL, JLTVILGVLDLIEAMASNGETLEKITNSRPPCVJLTVILGVL
co co amino acids 154-162 0f100PmeIl7 DLIEAIRRASNGETLEKITNSRPPCVKTWGQYWQV, uc7, co KTWGQYWQVDLIEAIRRASNGETLEKITNSRPPC'VKTWGQYWQV
"
tsi-) amino acids 209-217 of gplOOP'117 DLIEAIRRASNGETLEKITNSRPP CVITD
QVPF S V, 0 ITDQVPFSVDLIEAIRRASNGETLEKITNSRPPCVITD QVPFSV
amino acids 280-288 of gp10 OPmel 17 DLIEAIRRASNGETLEKITNSRPPCVYLEPGPVTA, 0 c7, YLEPGPVTADLIEAIRRASNGETLEKITNSRPPCVYLEPGPVTA
amino acids 457-466 of gp100Pmel"
DLIEAIRRASNGETLEKITNSRPPCVLLDGTATLRL, LLD GTATLRLDLIEAIRRASNGETLEKITNSRPP CVLID GTATLRL
amino acids 476-485 of gp1001'117 DLIEAIRRASNGETLEKITNSRPPCVVLYRYGSFSV, VLYRYGSFSVDLIEAIRRASNGETLEKITNSRPPCVVLYRYGSFSV
amino acids 301-309 of FRAME DLIEAIRRASNGETLEICITNSRPP CVLYVD
SLFFL, LYVD SLFFLDLIEAIRRASNGETLEKITNSRPPCVLYVD SLFFL
amino acids 292-303 of MAGE-6 DLIEALRRASNGETLEKITNSRPPCVKIS
GGPRISYPL, KIS GGPRISYPLD LLEAIRRASNGETLEKITNSRPP CVKIS GGPRISYPL
oe tµ.) tµ.) C35 Peptide/Epitope Exemplary Tumor Rejection Peptide Exemplary Polytopes amino acids 157-167 of NY-ESO-1 DLIEAIRRASNGETLEKITNSRPPCVSLIMWITQCFL, SLLMWITQCFLDLIEAIRRA.SNGETLEKITNSRPPCVSLLMWITQCFL
, tµ.) amino acids 157-165 of NY-ESO-1 DLIEAIRRASNGETLEKITNSRPPCVSLLMWITQC, oe SLLMWITQCDLIEAIRRASNGETLEICITNSRPPCVSLLMWITQC
amino acids 155-163 of NY-ESO-1 DLIEAIRRASNGETLEKITNSRPPCVQLSLLMWIT, QLSLLMWITDLIEAIRRASNGETLEKITNSRPPCVQLSLLMVVIT
amino acids 157-170 of NY-ESO-1 SLLMWITQCFLPVFDLIEAIRRASNGETLEKITNSRPPCVTSYVKVLHHMVKISG
and amino acids 281-295 of MAGE-3 P84 - V113 of SEQ ID NO:2 amino acids 161-169 of MAGE-1 PYEKDLIEAIRRASNGETLEKITNSRPPCVEADPTGHSY, PYEKDLIEAIRRASNGET

LEKITNSRPP CV
co amino acids 230-238 of MAGE-1 PYEKDLIEAIRRASNGETLEKITNSRPPCVSAYGEPRKL, co c7, op SAYGEPRKLPYEKDLIEAIRRASNGETLEKITNSRPPCVSAYGEPRKL

amino acids 168-176 of MAGE-3 PYEKDLIEAIRRASNGETLEKITNSRPPCVEVDPIGHLY, 0 EVDPIGHLYPYEKDLIEAIRRASNGETLEKITNSRPPCVEVDPIGHLY
amino acids 271-279 of MAGE-3 PYEKDLIEAIRRASNGETLEKITNSRPPCVFLWGPRALV, 0 c7, FLWGPRALVPYEKDLIEAIRRASNGETLEKITNSRPPCVFLWGPRALV
amino acids 167-176 of MAGE-3 PYEKDLIEAIRRASNGETLEKITNSRPPCVMEVDPIGHLY, MEVDPIGHLYPYEKDLIEAIRRA SNGETLEKITNSRPPCVMEVDPIGHLY
amino acids 2-10 of BAGE
PYEKDLIEMRRASNGETLEKITNSRPPCVAARAVFLAL, AARAVFLALPYEKDLIEAIRRASNGETLEKITNSRPPCVAARAVFLAL
amino acids 9-16 of GAGE-1,2 PYEKDLIEMRRASNGETLEKITNSRPPCVYRPRPRRY, YRPRPRRYPYEKDLIEAIRRASNGETLEKITNSRPPCVYRPRPRRY
amino acids 11-20 of RAGE
PYEKDLIEAIRRASNGETLEICITNSRPPCVSPSSNRIRNT, SP SSNRIRNTPYEKDLIEMRRASNGETLEKITNSRPPCVSP SSNRIRNT
tµ.) -tµ.) C35 Peptide/Epitope Exemplary Tumor Rejection Peptide Exemplary Polytopes amino acids 23-32 of CDK4 PYEKDLIEAIRRASNGETLEKITNSRPPCVARDPHSGHFV, ARDPHSGHFVPYEKDLLEAIRRASNGETLEKITNSRPPCV ARDPHSGHFV
tµ.) amino acids 29-37 of P-catenin PYEKDLIEAIRRASNGETLEKITNSRPPCVSYLDSGIEIS, oe SYLDSGIHSPYEKDLIEAIRRASNGETLEKITNSRPPCVSYLDSGITIS
amino acids 1-9 of tyrosinase PYEKDLIEAIRRASNGETLEKITNSRPPCVMLLAVLYCL, MLLAVLYCLPYEKDLIEAIRRASNGETLEKITNSRPPCVMLLAVLYCL
amino acids 206-214 of tyrosinase PYEKDLIEAIRRASNGETLEKITNSRPPCVAFLPWHRLF, AFLPWHRLFPYEKDLLEAIRRASNGETLEKITNSRPPCVAFLPWHRLF
amino acids 56-70 of tyrosinase PYEKDLIEAIRRASNGETLEKITNSRPPCVQNILLSNAPLGPQFP, QNILLSNAPLGPQFPPYEKDLLEAIRRASNGETLEKITNSRPPCVQNILLSNAPLGPQFP
co co amino acids 448-462 of tyrosinase PYEKDLIEA1RRASNGETLEKITNSRPPCVDYSYLQDSDPDSFQD, DYSYLQDSDPDSFQDPYEKDLIEA1RRASNGETLEKITNSRPPCVDYSYLQDSDPDSFQD u "
"

amino acids 32-40 of Me1an-AmART-1 PYEKDLIEAIRRASNGETLEKITNSRPPCVJLTVILGVL, 0 ILTVILGVLPYEKDLLEAIRRASNGETLEKITNSRPPCVILTVILGVL
amino acids 154-162 of gplOOP'117 PYEKDLIEAIRRASNGETLEKITNSRPPCVKTWGQYWQV, 0 KTWGQYWQVPYEKDLIEAIRRASNGETLEKITNSRPPCVKTWGQYWQV
amino acids 209-217 of gp1001"117 PYEKDLLEAIRRASNGETLEKITNSRPPCVITDQVPFSV, ITDQVPFSVPYEKDLIEAIRRASNGETLEKITNSRPPCVITDQVPFSV
amino acids 280-288 of gp100'117 PYEKDLIEAIRRASNGETLEKITNSRPPCVYLEPGPVTA, YLEPGPVTAPYEKDLIEMRRASNGETLEKITNSRPPCVYLEPGPVTA
amino acids 457-466 of gp100'117 PYEKDLIEAIRRASNGETLEKITNSRPPCVLLDGTATLRL, amino acids 476-485 of gp100'117 PYEKDLIEAIRRASNGETLEKITNSRPPCVVLYRYGSFSV, VLYRYGSFSVPYEKDLIEAIRRASNGETLEKITNSRPPCVVLYRYGSFSV
oe tµ.) tµ.) C35 Peptide/Epitope Exemplary Tumor Rejection Peptide Exemplary Polytopes amino acids 301-309 of PRAME
PYEKDLLEAIRRASNGETLEKITNSRPPCVLYVD SLFFL, LYVD SLFFLPYEKDLIEAIRRASNGETLEKITNSRPPCVLYVD SLFFL
tµ.) amino acids 292-303 of MAGE-6 PYEKDLEAIRRASNGETLEKITNSRPPCVKIS GGPRISYPL, KIS GGPRISYPLPYEKDLIEAIRRASNGETLEKITNSRPPCVKIS GGPRISYPL
amino acids 157-167 of NY-ESO-1 PYEKDLIEAIRRASNGETLEKITNSRPPCVSLLMWITQCFL, SLLMWITQCFLPYEKDLIEAIRRASNGETLEKITNSRPPCVSLLMWITQCFL
amino acids 157-165 of NY-ES 0-1 PYEKDLIEAIRRASNGETLEKITNSRPPCVSLLMWITQC, SLLMWITQCPYEKDLIEAIRRASNGETLEKITNSRPPCVSLLMWITQC
amino acids 155-163 of NY-ESO-1 PYEKDLIEAIRRASNGETLEKITNSRPPCVQLSLLMWIT, QLSLLMIATITPYEKDLIEAIRRASNGETLEKITNSRPPCVQLSLLMWIT

co amino acids 157-170 of NY-ES 0-1 SLLMWITQCFLPVFPYEKDLIEAIRRASNGETLEKITNSRPPCVTSYVKVLHHMVKISG co oo and amino acids 281-295 of MAGE-3 1\) K77 - V113 of SEQ ID N0:2 amino acids 161-169 of MAGE-1 KLENGGFPYEICDLIEAIRRASNGETLEICITNSRPPCVEADPTGHSY, 0 KLENGGFPYEKD LIEAIR
EADPTGHSYKLENGGFPYEICDLIEAIRRASNGETLEKITNSRPPCV EADPTGHSY
RA SNGETLEKITNSRPP CV
amino acids 230-238 of MAGE-1 KLENGGFPYEKDLIEAIRRASNGETLEKIINSRPPCVSAYGEPRKL, 0 c7, SAYGEPRKLKLENGGFPYEICDLIEAIRRASNGETLEKIINSRPPCVSAYGEPRKL
amino acids 168-176 of MAGE-3 KLENGGFPYEKDLIEAIRRASNGETLEKITNSRPPCVEVDPIGHLY, EVDPIGHLYKLENGGFPYEKDLIEAIRRASNGETLEICITNSRPPCVEVDPIGHLY
amino acids 271-279 of MAGE-3 KLENGGFPYEKDLIEAIRRASNGETLEICITNSRPPCVFLWGPRALV, FLWGPRALVKLENGGFPYEICDLIEAIRRASNGETLEKITNSRPPCVFLWGPRALV
amino acids 167-176 of MAGE-3 KLENGGFPYEKDLIEAIRRASNGETLEKITNSRPPCVMEVDPIGHLY, MEVDPIGHLYKLENGGFPYEKDLIEMRRASNGETLEICITNSRPPCVMEVDPIGHLY
amino acids 2-10 of BAGE
KLENGGFPYEKDLIEAIRRASNGETLEKITNSRPPCVAARAVFLAL, AARAVFLALKLENGGFPYEKDLIEAERRASNGETLEKITNSRPPCVAARAVFLAL
tµ.) tµ.) C35 Peptide/Epitope Exemplary Tumor Rejection Peptide Exemplary Polytopes amino acids 9-16 of GAGE-1,2 KLENGGFPYEKDLIEAIRRASNGETLEKITNSRPPCVYRPRPRRY, YRPRPRRYKLENGGFPYEKDLIEAIRRASNGETLEKITNSRPPCVYRPRPRRY
tµ.) amino acids 11-20 of RAGE KLENGGFPYEKD LIEAIRRA
SNGETLEKITNSRPPCVSPS SNRIRNT, SP S SNRIRNTKLENGGFPYEKDLIEAIRRASNGETLEKITNSRPPCVSPS SNRIRNT
amino acids 23-32 of CDK4 KLENGGFPYEKDLIEAIRRASNGETLEKITNSRPPCVARDPHS GHFV, ARDPHS GHFVKLENGGFPYEKDLIEAERRASNGETLEKITNSRPPCVARDPHS GHFV
amino acids 29-37 off3-catenin KLENGGFPYEKDLIEAIRRASNGETLEKITNSRPPCVSYLDS GIHS, SYLD S GIHSKLENGGFPYEKDLIEAIRRASNGETLEKITNSRPPCVSYLD S GIBS
amino acids 1-9 of tyrosinase KLENGGFPYEKDLIEAIRRASNGETLEKITNSRPPCVMLLAVLYCL, MLLAVLYCLKLENGGFPYEKDLIEAIRRASNGETLEKIINSRPPCVMLLAVLYCL

co amino acids 206-214 of tyrosinase KLENGGFPYEKDLIEAIRRASNGETLEKITNSRPPCVAFLPWHRLF, co 0-, AFLPWHRLFKLENGGFPYEKDLIEAIRRASNGETLEKITNSRPPCVAFLPWHRLF
ul ci;) amino acids 56-70 of tyrosinase KLENGGFPYEKDLIEAIRRASNGETLEKITNSRPPCVQNILLSNAPLGPQFP, 0 QNILLSNAPLGPQFPKLENGGFPYEKDLIEAIRRASNGETLEKITNSRPPCVQNILLSNAPLG
PQFP

c7, amino acids 448-462 of tyrosinase KLENGGFPYEKDLIEAIRRASNGETLEKITNSRPPCVDYSYLQDSDPDSFQD, DYSYLQD SD PD SF QDKLENGGFPYEKDLIEAIRRASNGETLEKIINSRPP CVDYSYLQD SD
PD SFQD
amino acids 32-40 of Me1an-AmART1 KLENGGFPYEKDLIEAIRRASNGETLEKITNSRPPCVJLTVILGVL, JLTVILGVLKLENGGFPYEKDLIEAIRRASNGETLEKITNSRPPCVJLTVILGVL
amino acids 154-162 of gp100'17 KLENGGFPYEKDLIEAIRRASNGETLEKITNSRPPCVKTWGQYWQV, KTWGQYWQVKLENGGFPYEKDLIEAIRRASNGETLEKITNSRPP CVKTWGQYWQV
amino acids 209-217 of gp100Pme117 KLENGGFPYEKDLIEAIRRASNGETLEKITNSRPPCVILITDQVPFSV, ITD QVPFSVKLENGGFPYEKDLIEAIRRASNGETLEKITNSRPP CVILITD QVPFSV
oe tµ.) tµ.) C35 Peptide/Epitope Exemplary Tumor Rejection Peptide Exemplary Polytopes amino acids 280-288 of gp100P'117 KLENGGFPYEKDLIEMRRASNGETLEKITNSRPPCVYLEPGPVTA, YLEPGPVTAKLENGGFPYEKDLIEAIRRASNGETLEKITNSRPPCVYLEPGPVTA
tµ.) amino acids 457-466 of gp100P'117 KLENGGFPYEKDLLEAIRRASNGETLEKITNSRPPCVLLDGTATLRL, LLDGTATLRLICLENGGFPYEKDLIEAIRRASNGETLEKITNSRPPCVLLDGTATLRL
amino acids 476-485 of gp100Pme117 KLENGGFPYEKDLIEAIRRASNGETLEKITNSRPPCVVLYRYGSFSV, VLYRYGSFSVKLENGGFPYEKDLIEAIRRASNGETLEICITNSRPPCVVLYRYGSFSV
amino acids 301-309 of PRAME
KLENGGFPYEKDLIEAIRRASNGETLEKITNSRPPCVLYVDSLFFL, LYVDSLFFLKLENGGFPYEKDLIEAIRRASNGETLEKITNSRPPCVLYVDSLFFL
amino acids 292-303 of MAGE-6 KLENGGFPYEKDLIEAIRRASNGETLEKITNSRPPCVKISGGPRISYPL, KISGGPRISYPLKLENGGFPYEKDLIEAIRRASNGETLEKITNSRPPCVKISGGPRISYPL

co amino acids 157-167 of NY-ESO-1 KLENGGFPYEKDLIEAIRRASNGETLEKITNSRPPCVSLLMWITQCFL, co c7, 1--µ
SLLMWITQCFLKLENGGFPYEKDLIEMRRASNGETLEKITNSRPPCVSLLMWITQCFL
amino acids 157-165 of NY-ESO-1 KLENGGFPYEKDLIEAIRRASNGETLEKITNSRPPCVSLLMWITQC, 0 SLLMWITQCKLENGGFPYEKDLIEAIRRASNGETLEKITNSRPPCVSLLIVIWITQC
amino acids 155-163 of NY-ESO-1 KLENGGFPYEKDLIEAIRRASNGETLEICITNSRPPCVQLSLLMWIT, 0 c7, QLSLLMWITKLENGGFPYEKDLIEAIRRASNGETLEICITNSRPPCVQLSLLMWIT
amino acids 157-170 of NY-ES 0-1 SLLMWITQCFLPVFKLENGGFPYEKDLIEAIRRASNGETLEICITNSRPPCVTSYVKVLHHM
and amino acids 281-295 of MAGE-3 VKISG
= Q72-V113 of SEQ ID N0:2 amino acids 161-169 of MAGE-1 OLVFSKLENGGFPYEKDLIEAIRRASNGETLEKITNSRPPCVEADPTGHSY, QLVFSKLENGGFPYEKD
EADPTGHSYQLVFSICLENGGFPYEKDLIEAIRRASNGETLEICITNSRPPCVEADPTGHSY
LIEAIRRASNGETLEKITN
SRPPCV amino acids 230-238 of MAGE-1 OLVFSKLENGGFPYEKDLIEAIRRASNGETLEKITNSRPPCVSAYGEPRKL, SAYGEPRKLOLVFSKLENGGFPYEKDLIEAIRRASNGETLEKITNSRPPCVSAYGEPRKL
amino acids 168-176 of MAGE-3 QLVFSKLENGGFPYEKDLIEA1RRASNGETLEKITNSRPPCVEVDPIGHLY, EVDPIGHLYOLVFSKLENGGFPYEKDLIEAIRRASNGETLEKITNSRPPCVEVDPIGHLY
oe tµ.) tµ.) =
C35 Peptide/Epitope Exemplary Tumor Rejection Peptide Exemplary Polytopes amino acids 271-279 of MAGE-3 QLVFSKLENGGFPYEKDLIEAIRRASNGETLEKITNSRPPCVFLWGPRALV, FLWGPRALVOLVFSKLENGGFPYEKDLIEAIRRASNGETLEKITNSRPPCVFLWGPRALV
tµ.) amino acids 167-176 of MAGE-3 QLVF'SKLENGGFPYEKDLIEAIRRASNGETLEKITNSRPPCVMEVDPIGHLY, MEVDPIGHLYQLVFSKLENGGFPYEKDLIEAIRRASNGETLEKITNSRPPCVMEVDPIGHLY
amino acids 2-10 of BAGE
QLVFSKLENGGFPYEKDLIEAIRRASNGETLEKITNSRPPCVAARAVFLAL, AARAVFLALQLVFSKLENGGFPYEKDLIEAIRRASNGETLEKITNSRPPCVAARAVFLAL
amino acids 9-16 of GAGE-1,2 QLVFSKLENGGFPYEKDLIEAIRRASNGETLEKITNSRPPCVYRPRPRRY, YRPRPRRYQLVFSKLENGGFPYEKDLIEAIRRASNGETLEKITNSRPPCVYRPRPRRY
amino acids 11-20 of RAGE QLVF
SKLENGGFPYEKDLIEAIRRASNGETLEKITNSRPPCVSPSSNRIRNT, SPSSNRIRNTQLVFSKLENGGFPYEKDLIEAIRRASNGETLEKITNSRPPCVSPSSNRIRNT

co amino acids 23-32 of CDK4 QLVFSKLENGGFPYEKDLIEAIRRASNGETLEKITNSRPPCVARDPHSGHFV, co c7, ARDPHSGHFVQLVFSKLENGGFPYEKDLIEAIRRASNGETLEKITNSRPPCVARDPHSGHFV
OQ
amino acids 29-37 of p-catenin QLVFSKLENGGFPYEKDLIEAIRRASNGETLEKIINSRPPCVSYLDSGIHS, 0 SYLDSGIHSQLVFSKLENGGFPYEKDLIEAIRRASNGETLEKIINSRPPCVSYLDSGIHS
amino acids 1-9 of tyrosinase QLVFSKLENGGFPYEKDLIEAIRRASNGETLEKITNSRPPCVMLLAVLYCL, 0 c7, amino acids 206-214 of tyrosinase QLVFSKLENGGFPYEKDLIEAIRRASNGETLEKITNSRPPCVAFLPWHRLF, AFLPWHRLFQLVFSKLENGGFPYEKDLIEAIRRASNGETLEKIINSRPPCVAFLPWHRLF
amino acids 56-70 of tyrosinase OLVFSICLENGGFPYEKDLIEAIRRASNGETLEKITNSRPPCVQNILLSNAPLGPQFP, QNILLSNAPLGPQFPQLVFSKLENGGFPYEKDLIEAIRRASNGETLEKITNSRPPCVQNILLS
NAPLGPQFP
oe JI

C35 Peptide/Epitope Exemplary Tumor Rejection Peptide Exemplary Polytopes amino acids 448-462 of tyrosinase QLVFSKLENGGFPYEKDLIEAIRRASNGETLEKITNSRPPCVDYSYLQD SDPD SF QD , DYSYLQD SDPD SF QD QLVFSKLENGGFPYEKDLIEAIRRASNGETLEKITNSRPPCVDYSY
'lotto LQD SDPD SFQD
amino acids 32-40 of Me1an-AmAkT4 QLVFSKLENGGFPYEKDLLEAIRRASNGETLEKITNSRPPCVJLTVILGVL, JLTVILGVLQLVFSKLENGGFPYEKDLLEAIRRASNGETLEKITNSRPPCVJLTVILGVL
amino acids 154-162 of gp 10 OPme 1 17 QLVF
SKLENGGFPYEKDLIEAIRRASNGETLEKITNSRPP CVKTWGQYWQV, KTWGQYWQVQLVFSKLENGGFPYEKDLIEAIRRASNGETLEICITNSRPPCVKTWGQYWQV
amino acids 209-217 of gp 1001'117 QLVFSKLENGGFPYEKDLIEAIRRASNGETLEKITNSRPPCVITD QVPFSV, 0 ITD QVPF S VQLVF SKLENGGFP YEKDLIEAIRRASNGETLEKITNSRPP CVITD QVPFSV
co co amino acids 280-288 of gp100'117 QLVFSKLENGGFPYEKDLIEAIRRASNGETLEKITNSRPPCVYLEPGPVTA, c7, I = =
,OD
YLEP GP VTAQLVF SKLENGGFP YEKDLIEAIRRASNGETLEKFINSRPPCVYLEP GP VTA

amino acids 457-466 of gp 100'117 Q LVF SKLENGGFP
YEKDLIEAIRRASNGETLEICITNS RPP CVLLD GTATLRL, LLD GTATLRLQLVFSKLENGGFPYEKDLIEMRRASNGETLEKITNSRPP CVLLD GTATLRL

amino acids 476-485 of gp100'17 QLVFSKLENGGFPYEKDLIEAIRRASNGETLEKITNSRPPCVVLYRYGSFSV, c7, VLYRYGSFSVQLVFSKLENGGFPYEKDLIEAIRRASNGETLEKITNSRPPCVVLYRYGSFSV
amino acids 301-309 of PRAME
QLVFSKLENGGFPYEKDLLEAIRRASNGETLEKITNSRPPCVLYVD SLFFL, LYVD SLFFLQLVFSKLENGGFP YEKDLIEAIRRASNGETLEKITNSRPP CVLYVD SLFFL
amino acids 292-303 of MAGE-6 QLVFSKLENGGFPYEKDLIEAIRRASNGETLEKITNSRPPCVKISGGPRISYPL, amino acids 157-167 of NY-ES 0-1 QLVF
SKLENGGFPYEKDLIEA1RRASNGETLEKITNSRPP CVSLLMWITQCFL, SLLMWITQCFLOLVFSKLENGGFPYEKDLIEAIRRASNGETLEKITNSRPPCVSLLMWITQCFL re C35 Peptide/Epitope Exemplary Tumor Rejection Peptide Exemplary Polytopes amino acids 157-165 of NY-ES 0-1 QLVFSICLENGGFPYEKDLIEAIRRASNGETLEKITNSRPPCVSLLMWITQC, SLLMWITQCOLVFSKLENGGFPYEKDLIEAIRRASNGETLEKITNSRPPCVSLLMWITQC
tµ.) amino acids 155-163 of NY-ESO-1 QLVFSKLENGGFPYEKDLIEAIRRASNGETLEKITNSRPPCVQLSLLMWIT, QLSLLMWITQLVFSKLENGGFPYEICDLLEAIRRASNGETLEKITNSRPPCVQLSLLMWIT
amino acids 157-170 of NY-ESO-1 SLLMWITQCFLPVFOLVFSICLENGGFPYEICDLIEAIRRASNGETLEKITNSRPPCVTSYVKV
and amino acids 281-295 of MAGE-3 LHHMVKISG
F65 to V113 of SEQ ID N0:2 amino acids 161-169 of MAGE-1 FEIENGQLVFSKLENGGFPYEKDLLEAIRRASNGETLEKITNSRPPCVEADPTGHSY, FEIEINGQLVFSKLENGGF
PYEKDLIEAIRRA SN GET
EADPTGHSYFEIEINGOLVFSICLENGGFPYEKDLIEAIRRASNGETLEKITNSRPPCVEADP
LEKITNSRPPCV TGHSY

amino acids 230-238 of MAGE-1 FEIEINGQLVFSKLENGGFPYEKDLIEAIRRASNGETLEKITNSRPPCVSAYGEPRKL, co co c7, co SAYGEPRKLFEIEINGQLVFSKLENGGFPYEKDLIEALRRASNGETLEKITNSRPPCVSAYG
"
EPRKL

amino acids 168-176 of MAGE-3 FEIEINGQLVFSKLENGGFPYEKDLIEAIRRASNGETLEKITNSRPPCVEVDPIGHLY, EVDPIGHLYFEIEINGQLVFSICLENGGFPYEKDLIEAIRRASNGETLEKITNSRPPCVEVDPI
c7, GHLY
amino acids 271-279 of MAGE-3 FEIEINGOLVFSICLENGGFPYEKDLIEAIRRASNGETLEKITNSRPPCVFLWGPRALV, FLWGPRALVFEIEINGQLVFSKLENGGFPYEKDLIEAIRRASNGETLEKITNSRPPCVFLWG
PRALV
amino acids 167-176 of MAGE-3 FEIEINGOLVFSKLENGGFPYEKDLIEAIRRASNGETLEKITNSRPPCVMEVDPIGHLY, MEVDPIGHLYFEIEINGOLVFSKLENGGFPYEICDLIEAIRRASNGETLEICITNSRPPCVMEV
DPIGHLY
oe tµ.) tµ.) C35 Peptide/Epitope Exemplary Tumor Rejection Peptide Exemplary Polytopes amino acids 2-10 of BAGE
FEIEINGQLVFSKLENGGFPYEKDLIEAIRRASNGETLEKITNSRPPCVAARAVFLAL, AARAVFLALFEIEINGQ LVF SKLENGGFPYEKD LIEAIRRA SNGETLEKITNSRPP CVAARA
'lotto VFLAL
amino acids 9-16 of GAGE-1,2 FEIEINGOLVFSICLENGGFPYEKDLLEAIRRASNGETLEKITNSRPPCVYRPRPRRY, YRPRPRRYFEIE]NGQLVFSKLENGGFPYEKDLIEAIRRASNGETLEKITNSRPPCVYRPRPRRY
amino acids 11-20 of RAGE FEIEINGQLVF
SKLENGGFPYEKDLIEAIRRA SNGETLEKITNSRPP CVSP S SNRIRNT, SP S SNRIRNTFEIEINGQLVFSICLENGGFPYEKDLIEAIRRASNGETLEKITNSRPPCVSPS SN
RIRNT

co amino acids 23-32 of CDK4 FEIEINGQLVFSICLENGGFPYEKDLIEAIRRASNGETLEKITNSRPPCVARDPHS GHFV, co c7, co "
ARDPHS GHFVFEIEINGQLVFSKLENGGFPYEKDLIEAIRRASNGETLEKITNSRPPCVARD

PHS GHFV

amino acids 29-37 off3-catenin FEIEINGQLVFSKLENGGFPYEKDLIEAIRRASNGETLEICITNSRPPCVSYLD S GIHS, =

c7, SYLD S GIHSFEIEINGQ LVF SKLENGGFPYEKDLIEAIRRA SNGETLEKITNSRPP CVSYLD S
. GIHS
amino acids 1-9 of tyrosinase FEIEINGOLVFSKLENGGFPYEKDLIEAIRRASNGETLEKITNSRPPCVMLLAVLYCL, VLYCL
amino acids 206-214 of tyrosinase FEIE1NGQLVFSKLENGGFPYEKDLIEAMRASNGETLEKITNSRPPCVAFLPWHRLF, AFLPWHRLFFEIEINGQLVFSKLENGGFPYEKDLIEAIRRA SNGETLEKITNSRPP CVAFLP
ccg WHRLF
oe tµ.) c.;11 tµ.) C35 Peptide/Epitope Exemplary Tumor Rejection Peptide Exemplary Polytopes amino acids 56-70 of tyrosinase FEIEINGQLVFSICLENGGFPYEKDLLEAIRRASNGETLEKITNSRPPCVQNILLSNAPLGPQFP, =
QNILLSNAPLGPQFPFEIEINGOLVFSKLENGGFPYEKDLIEAIRRASNGETLEKIINSRPK 'lotto VQNILLSNAPLGPQFP
amino acids 448-462 of tyrosinase FEIEINGOLVFSKLENGGFPYEICDLIEAIRRASNGETLEKITNSRPPCVDYSYLQDSDPDSFQD, DYSYLQD SDPD S F QD FEIEINGQ LVF SKLENGGFPYEKDLIEAIRRASNGETLEICITNSRPP
CVDYSYLQDSDPD SFQD
amino acids 32-40 of Melan-AmART-I
FEIEINGQLVFSKLENGGFPYEKDLIEAIRRASNGETLEKITNSRPPCVJLTVILGVL, JLTVILGVLFELDNGQLVFSKLENGGFPYEKDLIEAIRRASNGETLEKITNSRPPMLTVILGVL

co amino acids 154-162 of gp100'117 FEIEINGQLVESKLENGGFPYEKDLIEAIRRASNGETLEKITNSRPPCVKTWGQYWQV, co c7, i=¨=
co KTWGQYWQVFEIEINGQLVFSKLENGGFPYEKDLIEAIRRASNGETLEKITNSRPPCVKTW tcP
(ND) GQYWQV

amino acids 209-217 of gp 1 00P'117 FELEINGQLVFSKLENGGFPYEKDLIEAIRRASNGETLEICITNSRPPCVITDQVPFSV, c7, ITD QVPF S VFEIEINGQLVF SKLENGGFPYEICDLIEAIRRASNGETLEKITNSRPP CVITD QV
PFSV
amino acids 280-288 of gp100'117 FEIEINGQLVFSKLENGGFPYEKDLIEAIRRASNGETLEKITNSRPPCVYLEPGPVTA, YLEPGPVTAFEIEINGOLVFSKLENGGFPYEKDLIEAIRRASNGETLEICITNSRPPCVYLEPG
PVTA
amino acids 457-466 of gp100'117 FEIEINGQLVESKLENGGFPYEKDLIEAIRRASNGETLEKITNSRPP CVLLD GTATLRL, LLD GTATLRLFEIEINGQLVESKLENGGFPYEKDLIEAIRRASNGETLEICITNSRPPCVLLD G
TATLRL
oe tµ.) tµ.) C35 Peptide/Epitope Exemplary Tumor Rejection Peptide Exemplary Polytopes amino acids 476-485 of gp 1 OOPmehhl FEIEINGOLVFSKLENGGFPYEKDLIEAIRRASNGETLEKITNSRPPCVVLYRYGSFSV, VLYRYGSFSVFEIEINGQLVFSKLENGGFPYEKDLIEAIRRASNGETLEKITNSRPPCVVLY
tµ.) RYGSFSV
oe amino acids 301-309 of PRAME
FEIEINGQLVFSKLENGGFPYEKDLIEAIRRASNGETLEKITNSRPPCVLYVD SLFFL, LIND SLFFLFEIEINGOLVF SKLENGGFPYEKD LIEAIRRA SNGETLEKITNSRPP CVLYVD S
LFFL
amino acids 292-303 of MAGE-6 FEIEINGOLVFSKLENGGFPYEKDLIEMRRASNGETLEKITNSRPPCVKISGGPRISYPL, KIS GGPRIS YP LFEIEINGOLVF SKLENGGFPYEKDLIEAIRRA SNGETLEKITNSRPP CVKIS

GGPRISYPL
co co amino acids 157-167 of NY-ES 0-1 FEIEINGQL'VFSICLENGGFPYEKDLIEAIRRASNGETLEKITNSRPPCVILSLLM'WITQCFL, =
SLLMWITQCFLFEIEINGQLVFSKLENGGFPYEKDLIEAIRRASNGETLEKITNSRPPCVSLL

MWITQCFL
amino acids 157-165 of NY-ES 0-1 FEIEINGOLVF
SKLENGGFPYEKDLIEAIRRA SNGETLEKITNSRPP CVS LLMWITQ C, 0 c7, SLLMWITQCFEIEINGQLVFSKLENGGFPYEKDLIEAIRRASNGETLEKITNSRPPCVSLLM
WITQC
amino acids 155-163 of NY-ES 0-1 FEIEINGQLVFSKLENGGFPYEKDLIEAIRRASNGETLEKITNSRPPCVQLSLLMWIT, QLSLLMWITFEIEINGOLVFSKLENGGFPYEKDLIEAIRRASNGETLEKITNSRPP CVQLSLL
MWIT

amino acids 157-170 of NY-ESO-1 SLLMWITQCFLPVFFEIEINGOLVFSKLENGGFPYEKDLIEAIRRASNGETLEKITNSRPPCV
and amino acids 281-295 of MAGE-3 TSYVKVLHHMVKISG
oe tµ.) C35 Peptide/Epitope Exemplary Tumor Rejection Peptide Exemplary Polytopes L59 - V113 of SEQ ID NO:2 amino acids 161-169 of MAGE-1 LGGTGAFEIEINGOLVFSKLENGGFPYEKDLIEAIRRASNGETLEKITNSRPPCVEADPTGHSY, LGGTGAFEIEINGQLVFS
KLENGGFPYEKDLIEAIR
EADPTGHSYLGGTGAFEIEINGOLVFSKLENGGFPYEICDLIEAIRRASNGETLEKITNSRPP
oe RASNGETLEKI'TNSRPPCV CVEADPTGHSY
amino acids 230-238 of MAGE-1 LGGTGAFEIEINGOLVFSKLENGGFPYEKDLIEAIRRASNGETLEKITNSRPPCVSAYGEPRKL, SAYGEPRKLLGGTGAFEIEINGQLVFSKLENGGFPYEKDLIEAIRRASNGETLEKITNSRPP
CVSAYGEPRKL
amino acids 168-176 of MAGE-3 LGGTGAFEIEINGOLVFSKLENGGFPYEKDLIEAIRRASNGETLEKITNSRPPCVEVDPIGHLY, EVDPIGHLYLGGTGAFEIElNGOLVFSKLENGGFPYEKDLIEAIRRASNGETLEKITNSRPPC

VEVDPIGHLY
co co amino acids 271-279 of MAGE-3 LGGTGAFEIEINGOLVFSKLENGGFPYEKDLMAIRRASNGETLEKTINSRPPCVFLWGPRALV, ED
FLWGPRALVLGGTGAFEIEINGOLVFSKLENGGFPYEKDLIEAIRRASNGETLEKITNSRPP

CVFLWGPRALV
amino acids 167-176 of MAGE-3 c7, HLY, MEVDPIGHLYLGGTGAFEIEINGQLVFSICLENGGFPYEKDLIEAIRRASNGETLEKITNSRP
PCVMEVDPIGHLY
amino acids 2-10 of BAGE
LGGTGAFEIEINGQLVFSICLENGGFPYEKDLIEAIRRASNGETLEKITNSRPPCVAARAVFLAL, AARAVFLALLGGTGAFEIEINGOLVFSKLENGGFPYEKDLIEAIRRASNGETLEICITNSRPP
CVAARAVFLAL

oe tµ.) tµ.) C35 Peptide/Epitope Exemplary Tumor Rejection Peptide Exemplary Polytopes amino acids 9-16 of GAGE-1,2 LGGTGAFEMINGQLVFSKLENGGFPYEKDLIEAIRRASNGETLEKITNSRPPCVYRPRPRRY, YRPRPRRYLGGTGAFEIEINGOLVFSKLENGGFPYEKDLIEAIRRASNGETLEKITNSRPPC
VYRPRPRRY
oe amino acids 11-20 of RAGE
LGGTGAFEIEINGOLVFSKLENGGFPYEKDLIEAIRRASNGETLEKITNSRPPCVSPSSNRIRNT, CVSPSSNRIRNT
amino acids 23-32 of CDK4 HFV, ARDPHSGHFVLGGTGAFEIEINGOLVFSKLENGGFPYEKDLIEAIRRASNGETLEKITNSRP
PCVARDPHSGHFV
c7, amino acids 29-37 off3-catenin LGGTGAFEIDNGOLVFSKLENGGFPYEKDLIEAIRRASNGETLEKITNSRPPCVSYLDSGIHS, "

SYLDSGIHSLGGTGAFEIEINGQLVFSKLENGGFPYEKDLIEAIRRASNGETLEKITNSRPPC
VSYLDSGIHS

amino acids 1-9 of tyrosinase LGGTGAFEIBINGOLVFSKLENGGFPYEKDLIEAIRRASNGETLEKITNSRPPCVMLLAVLYCL, c7, MLLAVLYCLLGGTGAFEIEINGQLVFSKLENGGFPYEKDLIEAIRRASNGETLEKITNSRPP
CVMLLAVLYCL
amino acids 206-214 of tyrosinase LGGTGAFEIEINGOLVFSKLENGGFPYEKDLIEALRRASNGETLEKITNSRPPCVAFLPWHRLF, AFLPWHRLFLGGTGAFEIEINGOLVESKLENGGFPYEKDLIEAIRRASNGETLEKITNSRPP ole CVAFLPWHRLF
oe C35 Peptide/Epitope Exemplary Tumor Rejection Peptide Exemplary Polytopes amino acids 56-70 of tyrosinase LGGTGAFEIEINGQLVFSKLENGGFPYEKDLIEAIRRASNGETLEKITNSRPPCVQNILLSNA
PLGPQFP, QNILLSNAPLGPQFPLGGTGAFEIEINGOLVFSKLENGGFPYEKDLIEAIRRASNGETLEKIT
NSRPPCVQNILLSNAPLGPQFP
amino acids 448-462 of tyrosinase LGGTGAFEIEINGOLVFSKLENGGFPYEKDLIEALR.RASNGETLEICITNSRPPCVDYSYLQD
SDPDSFQD, =
TNSRPPCVDYSYLQDSDPDSFQD
amino acids 32-40 of Melan-A' LGGTGAFEIEINGQLVFSKLENGGFPYEKDLIEAIRRASNGETLEICITNSRPPCVJLTVILGVL, JLTVILGVLLGGTGAFELEINGOLVFSKLENGGFPYEICDLIEAIRRASNGETLEKITNSRPPC , c7, VJLTVILGVL

amino acids 154-162 of gp100Pme117 WQV, KTWGQYWQVLGGTGAFEIEINGQLVESKLENGGFPYEKDLIEAIRRASNGETLEICITNSRP

c7, PCVKTWGQYWQV
amino acids 209-217 of gp1001"117 LGGTGAFEIEINGQLVFSKLENGGFPYEKDLIEALRRASNGETLEICITNSRPPCVITDQVPFSV, ITDQVPFSVLGGTGAFEIEINGQLVFSKLENGGFPYEKDLLEAIRRASNGETLEKITNSRPPC
VITDQVPFSV
amino acids 280-288 of gp100'117 LGGTGAFEIONGQLVFSKLENGGFPYEKDLIEAJRRASNGETLEKITNSRPPCVYLEPGPVTA, YLEPGPVTALGGTGAFEIEINGQLVFSICLENGGFPYEKDLIEAIRRASNGETLEKITNSRPP c7, CVYLEPGPVTA
oe C35 Peptide/Epitope Exemplary Tumor Rejection Peptide Exemplary Polytopes amino acids 457-466 of gp1001"117 LGGTGAFEIEINGQLVFSKLENGGFPYEKDLIEAIRRASNGETLEKITNSRPPCVLLDGTAT
LRL, tµ.) LLD GTATLRLLGGTGAFEIEINGQLVFSKLENGGFPYEKDLIEAIRRASNGETLEKITNSRP
P CVLLD GTATLRL
amino acids 476-485 of gp 1001me 1 1 LG GTGAFEIEING QLVF SKLENG
GFPYEKD LIEAIRRASNGETLEKITNSRPP CVVLYRYGS
FSV, VLYRYGSFSVLGGTGAFEIEINGQLVFSKLENGGFPYEKDLIEAIRRASNGETLEKITNSRP
P CVVLYRYGSFSV
amino acids 301-309 of PRAME
LGGTGAFETEINGQLVFSKLENGGFPYEKDLIEMRRASNGETLEKITNSRPPCVLYVDSLFFL, co LYVD SLFFLLGGTGAFEMINGQLVFSKLENGGFPYEKDLIEArRRASNGETLEKUNSRPPC , co c7, VLYVD SLFFL
co cr, amino acids 292-303 of MAGE-6 SYPL, KIS GGPRISYPLLGGTGAFEIEINGOLVFSKLENGGFPYEKDLIEAIRRASNGETLEKITNSR

c7, PP CVKIS GGPRISYPL
amino acids 157-167 of NY-ESO-1 LGGTGAFEIEINGQLVFSKLENGGFPYEICDLIEAIRRASNGETLEKITNSRPPCVSLLMWIT
QCFL, SLLMWITQCFLLGGTGAFEIEINGOLVFSKLENGGFPYEKDLIEAIRRASNGETLEKITNSR
PPCVSLLMWITQCFL
amino acids 157-165 of NY-ES 0-1 LGGTGAFEIEINGOLVFSKLENGGFPYEKDLIEAIRRASNGETLEKITNSRPPCVSLLMWITQC, 1-3 SLLMWITQ CLGGTGAFETErNGOLVF S1CLENGGFPYEKDLIEAIRRASNGETLEKITNSRPP
CVSLLMWITQC
oe t=.) tµ.) C35 Peptide/Epitope Exemplary Tumor Rejection Peptide Exemplary Polytopes amino acids 155-163 of NY-ES 0-1 LGGTGAFEIEINGQLVFSICLENGGFPYEKDLIEAIRRASNGETLEKITNSRPPCVQLSLLMWIT, QLSLLMWITLGGTGAFEIEINGOLVFSKLENGGFPYEKDLIEAIRRASNGETLEKITNSRPP
CVQLSLLMWIT
oo amino acids 157-170 of NY-ESO-1 SLLMWITQCFLPVFLGGTGAFEIEINGOLVFSKLENGGFPYEKDLLEAIRRASNGETLEICIT
and amino acids 281-295 of MAGE-3 NSRPPCVTSYVKVLHHMVKISG
099 -V113 of SEQ ID NO:2 amino acids 161-169 of MAGE-1 GETLEKITNSRPPCVEADPTGHSY, GETLEKITNSRPPCV
EADPTGHSYGETLEKITNSRPPCVEADPTGHSY
amino acids 230-238 of MAGE-1 GETLEKITNSRPPCVSAYGEPRKL, SAYGEPRKLGETLEKITNSRPPCVSAYGEPRKL

amino acids 168-176 of MAGE-3 GETLEKITNSRPPCVEVDPIGHLY, co co EVDPIGHLYGETLEKITNSRPPCVEVDPIGHLY

CO
ON
1\.) amino acids 271-279 of MAGE-3 GETLEICITNSRPPCVFLWGPRALV, FLWGPRALVGETLEKITNSRPPCVFLWGPRALV

amino acids 167-176 of MAGE-3 GETLEKITNSRPPCVMEVDPIGHLY, MEVDPIGHLYGETLEICTINSRPPCV_MEVDPIGHLY

amino acids 2-10 of BAGE GETLEKITNSRPPCVAARAVFLAL, AARAVFLALGETLEKITNSRPPCVAARAVFLAL
amino acids 9-16 of GAGE-1,2 GETLEKITNSRPPCVYRPRPRRY, YRPRPRRYGETLEKITNSRPPCVYRPRPRRY
amino acids 11-20 of RAGE GETLEICITNSRPPCVSPSSNRIRNT, SPSSNRIRNTGETLEICITNSRPPCVSPSSNRIRNT
amino acids 23-32 of CDK4 GETLEKITNSRPPCVARDPHSGHFV, ARDPHSGHFVGE'TLEICITNSRPPCVARDPHSGHFV
oo amino acids 29-37 of f3-catenin GETLEKITNSRPPCVSYLD S GIBS, SYLD S GIHSGETLEKITNSRPPCVSYLD S GIB S
amino acids 1-9 of Tyrosinase GETLEKITNSRPPCVMLLAVLYCL, tµ.) oe amino acids 206-214 of Tyrosinase GETLEKITNSRPPCVAFLPWHRLF, AFLPWHRLFGETLEKITNSRPPCVAFLPWHRLF
amino acids 56-70 of Tyrosinase GETLEKITNSRPP CVQNILLSNAPLGP QFP, QNILLSNAPLGPQFPGETLEKITNSRPPCVQNILLSNAPLGPQFP
amino acids 448-462 of Tyrosinase . GETLEKITNSRPPCVDYSYLQDSDPDSFQD, DYSYLQD SDPDSFQDGETLEKITNSRPPCVDYSYLQD SDPD SF QD
amino acids 32-40 of Melan-A' GETLEKITNSRPPCVJLTVILGVL, JLTVILGVLGETLEKITNSRPPCVJLTVILGVL
co co c7, op amino acids 154-162 of gplOOP'117 GETLEKITNSRPPCVKTWGQYWQV, c.N
KTWGQYWQVGETLEKITNSRPPCVKTWGQYWQV

amino acids 209-217 of gp100'17 SVAPPPEEVITDQVPFSV, ITDQVPFSVSVAPPPEEVITDQVPFSV

c7, amino acids 280-288 of gpi 0 oPmel 17 SVAPPPEEVYLEPGPVTA, YLEPGPVTASVAPPPEEVYLEPGPVTA
amino acids 457-466 of gp100P'117 GETLEKITNSRPPCVLLDGTATLRL, LLD GTATLRLGETLEKITNSRPPCVLLDGTATLRL
amino acids 476-485 of gp1001'117 GETLEKITNSRPPCVVLYRYGSFSV, VLYRYGSFSVGETLEKITNSRPPCVVLYRYGSFSV

amino acids 301-309 of PRAME GETLEKITNSRPPCVLYVD SLFFL, LYVD SLFFLGETLEKITNSRPPCVLYVD SLFFL
amino acids 292-303 of MAGE-6 GETLEKITNSRPPCVKISGGPRISYPL, tµ.) ; KIS GGPRISYPLGETLEKITNSRPPCVKIS GGPRISYPL
tµ.) amino acids 157-167 of NY-ESO-1 GETLEKITNSRPPCVSLLMWITQCFL, SLLMWITQCFLGETLEKITNSRPPCVSLLMWITQCFL
amino acids 157-165 of NY-ESO-1 GETLEKITNSRPPCVSLLMWITQC, SLLMWITQCGETLEKITNSRPPCVSLLMWITQC
t=.) oe = amino acids 155-163 of NY-ES0-1 GETLEKITNSRPPCVQLSLLMWIT, QLSLLMWITGETLEKITNSRPPCVQLSLLMWIT
amino acids 157-170 of NY-ES 0-1 SLLMWITQCFLPVFGETLEICITNSRPPCVTSYVKVLHHMVKISG
and amino acids 281-295 of MAGE-3 E100 -V113 of SEQ ID amino acids 161-169 of MAGE-1 ETLEICITNSRPPCVADPTGHSY, N0:2 EADPTGHSYETLEKITNSRPPCVEADPTGHSY
ETLEKITNSRPPCV

amino acids 230-238 of MAGE-1 ETLEICITNSRPPCVSAYGEPRKL, SAYGEPRICLETLEKITNSRPPCVSAYGEPRKL
co co c7, )--k op amino acids 168-176 of MAGE-3 ETLEKITNSRPPCVEVDPIGHLY, "
EVDPIGHLY ETLEKITNSRPPCVEVDPIGHLY
"

amino acids 271-279 of MAGE-3 ETLEKITNSRPPCVFLWGPRALV, FLWGPRALVETLEKITNSRPPCVFLWGPRALV

c7, amino acids 167-176 of MAGE-3 ETLEKITNSRPPCVMEVDPIGHLY, MEVDPIGHLYETLEKITNSRPPCVMEVDPIGHLY
amino acids 2-10 of BAGE ETLEKITNSRPPCVAARAVFLAL, AARAVFLALETLEKITNSRPPCVAARAVFLAL
amino acids 9-16 of GAGE-1,2 ETLEKITNSRPPCVYRPRPRRY, YRPRPRRYETLEKITNSRPPCVYRPRPRRY

amino acids 11-20 of RAGE ETLEKITNSRPP CVSPS SNRJRNT, SP S SNRIRNTETLEKITNSRPPCVSPS SNRIRNT
amino acids 23-32 of CDK4 ETLEKITNSRPPCVARDPHS GHFV, tµ.) ARDPHS GHFVETLEKITNSRPPCVARDPHS GHFV
tµ.) amino acids 29-37 of 13-catenin ETLEKITNSRPP CVSYLD SGIHS, SYLD S GIHSETLEKITNSRPPCVSYLD S GIHS
amino acids 1-9 of Tyrosinase ETLEKITNSRPPCVMLLAVLYCL, MLLAVLYCLETLEICIINSRPPCVMLLAVLYCL
tµ.) oe amino acids 206-214 of Tyrosinase ETLEKITNSRPPCVAFLPWHRLF, AFLPWHRLFETLEKITNSRPPCVAFLPWHRLF
amino acids 56-70 of Tyrosinase ETLEKITNSRPPCVQNILLSNAPLGPQFP, QNILLSNAPLGPQFPETLEKITNSRPPCVQNILLSNAPLGPQFP
amino acids 448-462 of Tyrosinase ETLEKITNSRPPCVDYSYLQDSDPDSFQD, DYSYLQD SDPD SFQDETLEKITNSRPPCVDYSYLQD SDPDSFQD

amino acids 32-40 of Melan-AmART-1 ETLEKITNSRPPCVILTVILGVL, JLTVILGVLETLEKITNSRPPCVJLTVILGVL
co co c7, amino acids 154-162 of gp100"117 ETLEKITNSRPPCVKTWGQYWQV, KTWGQYWQVETLEKITNSRPPCVKTWGQYWQV

amino acids 209-217 of gp100'117 ETLEKITNSRPPCVITDQVPFSV, EL
IIDQVPFSVETLEKITNSRPPCVITDQVPFSV

c7, amino acids 280-288 of gp100'117 ETLEICITNSRPPCVYLEPGPVTA, YLEPGPVTAETLEKITNSRPPCVYLEPGPVTA
amino acids 457-466 of gp100"117 ETLEKITNSRPPCVLLDGTATLRL, LLD GTATLRLETLEKITNSRPP CVLLD GTATLRL
amino acids 476-485 of gp100'117 ETLEKITNSRPPCVVLYRYGSFSV, VLYRYGSFSVETLEKITNSRPPCVVLYRYGSFSV

amino acids 301-309 of PRAME ETLEKITNSRPPCVLYVD SLFFL, LYVD SLFFLETLEKITNSRPP CVLYVDSLFFL
amino acids 292-303 of MAGE-6 ETLEKITNSRPPCVKISGGPRISYPL, tµ.) KIS GGPRISYPLETLEKITNSRPPCVKIS GGPRISYPL
tµ.) amino acids 157-167 of NY-ESO-1 ETLEKITNSRPPCVSLLMWITQCFL, SLLMWITQCFLETLEKTTNSRPPCVSLLMWITQCFL
amino acids 157-165 of NY-ESO-1 ETLEKITNSRPPCVSLLMWITQC, SLLMWITQCETLEKITNSRPPCVSLLMWITQC
tµ.) oe amino acids 155-163 of NY-ESO-1 ETLEKITNSRPPCVQLSLLMWIT, QLSLLMWITETLEKITNSRPPCVQLSLLMWIT
amino acids 157-170 of NY-ESO-1 SLLMWITQCFLPVFETLEKITNSRPPCVTSYVKVLHHMVKISG
and amino acids 281-295 of MAGE-3 co co c7, 1-=-i co 1\.) 1:71 oe TABLE C

=
C35 Epitope Analog Exemplary Tumor Rejection Peptide , Exemplary Polytope o .6.
.
.6.
Analog of S77 -Y85 of SEQ amino acids 161-169 of MAGE-1 KLENGGFPVEADPTGHSY, tµ.) oe ID NO:2 having valine EADPTGHSYKLENGGFPVEADPTGHSY
substituted for tyrosine at ninth amino acid residue amino acids 230-238 of MAGE-1 = KLENGGFPVSAYGEPRKL, (KLENGGFPV) SAYGEPRKLKLENGGFPVSAYGEPRKL
amino acids 168-176 of MAGE-3 KLENGGFPVEVDPIGHLY, EVDPIGHLYKLENGGFPVEVDPIGHLY
amino acids 271-279 of MAGE-3 KLENGGFPVFLWGPRALV, n FLWGPRALVKLENGGFPVFLWGPRALV

I.) amino acids 167-176 of MAGE-3 KLENGGFPVMEVDPIGHLY, co co MEVDPIGHLYKLENGGFPVMEVDPIGHLY
c7, co "
LP
I.) amino acids 2-10 of BAGE KLENGGFPVAARAVFLAL, AARAVFLALKLENGGFPVAARAVFLAL
a, H
IV
amino acids 9-16 of GAGE-1,2 KLENGGFPVYRPRPRRY, YRPRPRRYKLENGGFPVYRPRPRRY
c7, amino acids 11-20 of RAGE KLENGGFPVSPSSNRIRNT, .
SPSSNRIRNTKLENGGFPVSPSSNRIRNT
amino acids 23-32 of CDK4 KLENGGFPVARDPHSGHFV, , ARDPHSGHFVKLENGGFPVARDPHSGHFV
Iv n amino acids 29-37 of I3-catenin KLENGGFPVSYLDSGIHS, SYLDSGTEISKLENGGFPVSYLDSGIHS
cp o amino acids 1-9 of Tyrosinase KLENGGFPVMLLAVLYCL, 1--, oe MLLAVLYCLKLENGGFPVMLLAVLYCL
tµ.) un tµ.) C35 Epitope Analog Exemplary Tumor Rejection Peptide Exemplary Polytope amino acids 206-214 of Tyrosinase KLENGGFPVAFLPWHRLF, AFLPWHRLFICLENGGFPVAFLPWHRLF
tµ.) amino acids 56-70 of Tyrosinase KLENGGFPVQNILLSNAPLGPQFP, oe QNILLSNAPLGPQFPKLENGGFPVQNILLSNAPLGPQFP
amino acids 448-462 of Tyrosinase KLENGGFPVDYSYLQDSDPDSFQD, DYSYLQD SDPD SFQDKLENGGFPVDYSYLQD SDPD SF QD
amino acids 32-40 of Me1an-AmART4 KLENGGFPVJLTVILGVL, JLTVILGVLKLENGGFPVJLTVILGVL
amino acids 154-162 of gp100"117 KLENGGFPVKTWGQYWQV, KTWGQYVVQVICLENGGFPVKTWGQYWQV
co co amino acids 209-217 of gp10 OP' KLENGGFPVITDQVPFSV, c7, co ITDQVPFSVKLENGGFPVITD QVPF S V
i"

amino acids 280-288 of gp100"117 KLENGGFPVYLEPGPVTA, YLEPGPVTAKLENGGFPVYLEPGPVTA
amino acids 457-466 of gp100"117 KLENGGFPVLLDGTATLRL, c7, LLD GTATLRLKLENGGFPVLLDGTATLRL
amino acids 476-485 of gp100'117 KLENGGFPVVLYRYGSFSV, VLYRYGSFSVICLENGGFPVVLYRYGSFSV
amino acids 301-309 of PRAME KLENGGFPVLYVD SLFFL, LYVD SLFFLKLENGGFPVLYVD SLFFL
amino acids 292-303 of MAGE-6 KLENGGFPVKISGGPRISYPL, KIS GGPRISYPLKLENGGFPVKISGGPRISYPL
amino acids 157-167 of NY-ESO-1 KLENGGFPVSLLMWITQCFL, SLLMWITQCFLKLENGGFPVSLLMWITQCFLJI
oe tµ.) tµ.) C35 Epitope Analog Exemplary Tumor Rejection Peptide Exemplary Polytope amino acids 157-165 of NY-ESO-1 KLENGGFPVSLLMWITQC, SLLMWITQCKLENGGFPVSLLMWITQC
amino acids 155-163 of NY-ESO-1 KLENGGFPVQLSLLMWIT, tµ.) oe QLSLLMWITKLENGGFPVQLSLLMWIT
amino acids 157-170 of NY-ESO-1 and SLLMWITQCFLPVFKLENGGFPVTSYVKVLHHMVKISG
amino acids 281-295 of MAGE-3 Analog of K104 - C112 of amino acids 161-169 of MAGE-1 KITNSRPPLEADPTGHSY, SEQ ID N0:2 having leucine EADPTGIISYKITNSRPPLEADPTGHSY
substituted for cysteine at the ninth amino acid residue amino acids 230-238 of MAGE-1 KITNSRPPLSAYGEPRKL, (KITNSRPPL) SAYGEPRKLKITNSRPPLSAYGEPRKL

co amino acids 168-176 of MAGE-3 K1TNSRPPLEVDPIGHLY, co EVDPIGHLYKITNSRPPLEVDPIGHLY
c amino acids 271-279 of MAGE-3 KITNSRPPLFLWGPRALV, FLWGPRALVKITNSRPPLFLWGPRALV
amino acids 167-176 of MAGE-3 KITNSRPPLMEVDPIGHLY, MEVDPIGHLYKITNSRPPLMEVDPIGHLY
c7, amino acids 2-10 of BAGE KITNSRPPLAARAVFLAL, AARAVFLALKITNSRPPLAARAVFLAL
amino acids 9-16 of GAGE-1,2 KITNSRPPLYRPRPRRY, YRPRPRRYKITNSRPPLYRPRPRRY
amino acids 11-20 of RAGE KITNSRPPLSPSSNRIRNT, SPSSNRIRNTKITNSRPPLSPSSNRIRNT

amino acids 23-32 of CDK4 KITNSRPPLARDPHSGHFV, ARDPHSGHFVKITNSRPPLARDPHSGHFV
oe tµ.) tµ.) C35 Epitope Analog Exemplary Tumor Rejection Peptide Exemplary Polytope amino acids 29-37 off3-catenin KITNSRPPLSYLDSGIHS, SYLDSGIEISICITNSRPPLSYLDSGIHS
tµ.) amino acids 1-9 of Tyrosinase KITNSRPPLMLLAVLYCL, oe MLLAVLYCLKITNSRPPLMLLAVLYCL
amino acids 206-214 of Tyrosinase KITNSRPPLAFLPWHRLF, AFLPWHRLFICITNSRPPLAFLPWHRLF
amino acids 56-70 of Tyrosinase KITNSRPPLQNILLSNAPLGPQFP, QNILLSNAPLGPQFPKITNSRPPLQNILLSNAPLGPQFP
amino acids 448-462 of Tyrosinase KITNSRPPLDYSYLQDSDPDSFQD, DYSYLQDSDPDSFQDKITNSRPPLDYSYLQDSDPDSFQD

co amino acids 32-40 of Me1an-AmART-1 KITNSRPPLJLTVILGVL, rco JLTVILGVLKITNSRPPLILTVILGVL
9.`
amino acids 154-162 of gpl00Pme117 KITNSRPPLKTWGQYWQV, KTWGQYWQVKITNSRPPLKTWGQYWQV
amino acids 209-217 of gp100'17 KITNSRPPLITDQVPFSV, c7, ITDQVPFSVKITNSRPPLITDQVPFSV
amino acids 280-288 of gp100'17 KITNSRPPLYLEPGPVTA, YLEPGPVTAKITNSRPPLYLEPGPVTA
amino acids 457-466 of gp100'117 KITNSRPPLLLDGTATLRL, LLDGTATLRLKITNSRPPLLLDGTATLRL
amino acids 476-485 of gp100'117 KITNSRPPLVLYRYGSFSV, VLYRYGSFSVKITNSRPPLVLYRYGSFSV

amino acids 301-309 of PRAME KITNSRPPLLYVDSLFFL, LYVDSLFFLKITNSRPPLLYVDSLFFL
oe tµ.) tµ.) C35 Epitope Analog Exemplary Tumor Rejection Peptide Exemplary Polytope amino acids 292-303 of MAGE-6 KITNSRPPLICISGGPRISYPL, KIS GGPRISYPLKITNSRPPLKISGGPRISYPL
tµ.) amino acids 157-167 of NY-ESO-1 KITNSRPPLSLLM-VVITQCFL, oe SLLMWITQCFLKITNSRPPLSLLMWITQCFL
amino acids 157-165 of NY-ESO-1 KITNSRPPLSLLMWITQC, SLLMWITQCKITNSRPPLSLLMWITQC
amino acids 155-163 of NY-ESO-1 KITNSRPPLQLSLLMWIT, QLSLLMWITKITNSRPPLQLSLLMWIT
amino acids 157-170 of NY-ESO-1 SLLMWITQCFLPVFKITNSRPPLTSYVKVLHFIMVKISG
and amino acids 281-295 of MAGE-3 = 0 1.3 Analog of I105 - V113 of amino acids 161-169 of MAGE-1 ILNSRPPAVEADPTGHSY, c7, SEQ ID NO:2 having leucine EADPTGHSYLLNSRPPAVEADPTGHSY
substituted for threonine at the second amino acid amino acids 230-238 of MA GE-1 ILNSRPPAVSAYGEPRKL, 0 residue and alanine SAYGEPRKLILNSRPPAVSAYGEPRKL
substituted for cysteine at the amino acids 168-176 of MAGE-3 ILNSRPPAVEVDPIGHLY, 0 eighth amino acid residue c7, EVDPIGHLYIL,NSRPPAVEVDPIGHLY
(ILNSRPPAV) amino acids 271-279 of MAGE-3 ILNSRPPAVFLWGPRALV, FLWGPRALVILNSRPPAVFLWGPRALV
amino acids 167-176 of MAGE-3 ILNSRPPAVMEVDPIGHLY, MEVDPIGHLYILNSRPPAVMEVDPIGHLY
amino acids 2-10 of BAGE ILNSRPPAVAARAVFLAL, AARAVFLALILNSRPPAVAARAVFLAL
amino acids 9-16 of GAGE-1,2 ILNSRPPAVYRPRPRRY, YRPRPRRYILNSRPPAVYRPRPRRY
oe JI
tµ.) tµ.) C35 Epitope Analog Exemplary Tumor Rejection Peptide Exemplary Polytope amino acids 11-20 of RAGE ILNSRPPAVSPSSNRIRNT, SPSSNRIRNTILNSRPPAVSPSSNRIRNT
amino acids 23-32 of CDK4 ILNSRPPAVARDPHSGHFV, oe ARDPHSGHFVILNSRPPAVARDPHSGHFV
amino acids 29-37 of 13-catenin ILNSRPPAVSYLDSGIHS, SYLDSGIHS ILNSRPPAVSYLDSGIHS
amino acids 1-9 of Tyrosinase ILNSRPPAVMLLAVLYCL, MLLAVLYCLILNSRPPAVMLLAVLYCL
amino acids 206-214 of Tyrosinase LLNSRPPAVAFLPWHRLF, AFLPWHRLFILNSRPPAVAFLPWHRLF

amino acids 56-70 of Tyrosinase LLNSRPPAVQNILLSNAPLGPQFP, CO
c7, QNILLSNAPLGPQFPILNSRPPAVQNILLSNAPLGPQFP
amino acids 448-462 of Tyrosinase ILNSRPPAVDYSYLQDSDPDSFQD, DYSYLQDSDPDSFQDILNSRPPAVDYSYLQDSDPDSFQD
amino acids 32-40 of Me1an-AmART4 ILNSRPPAVJLTVILGVL, JLTVILGVLILNSRPPAVJLTVILGVL
c7, amino acids 154-162 of gp100'17 ILNSRPPAVKTWGQYWQV, KTWGQYWQVILNSRPPAVKTWGQYWQV
amino acids 209-217 of gp100' ILNSRPPAVITDQVPFSV, ITDQVPFSVILNSRPPAVITDQVPFSV
amino acids 280-288 of gp100'117 ILNSRPPAVYLEPGPVTA, amino acids 457-466 of gp1001'117 ILNSRPPAVLLDGTATLRL, LLDGTATLRLILNSRPPAVLLDGTATLRL
oe C35 Epitope Analog Exemplary Tumor Rejection Peptide Exemplary Polytope amino acids 476-485 of gplOOPmen7 ILNSRPPAVVLYRYGSFSV, VLYRYGSFSVILNSRPPAVVLYRYGSFSV
amino acids 301-309 of FRAME ILNSRPPAVLYVDSLFFL, tµ.) oe LYVDSLFFLILNSRPPAVLYVDSLFFL
amino acids 292-303 of MAGE-6 ILNSRPPAVKISGGPRISYPL, KISGGPRISYPLILNSRPPAVICISGGPRISYPL
amino acids 157-167 of NY-ESO-1 ILNSRPPAVSLLMWITQCFL, SLLMWITQCFLILNSRPPAVVSLLMWITQCFL
amino acids 157-165 of NY-ESO-1 ILNSRPPAVSLLMWITQC, SLLMWITQCILNSRPPAVSLLMWITQC

co amino acids 155-163 of NY-ESO-1 ILNSRPPAVQLSLLMWIT, co QLSLLMWITILNSRPPAVQLSLLMWIT
,õ
amino acids 157-170 of NY-ESO-1 SLLMWITQCFLPVF

and amino acids 281-295 of MAGE-3 Analog of I105 - V113 of amino acids 161-169 of MAGE-1 IMNSRPPAV EADPTGHSY, 0 c7, SEQ ID NO:2 having EADPTGHSYIMNSRPPAVEADPTGHSY
methionine substituted for threonine at the second amino acids 230-238 of MAGE-1 IMNSRPPAV SAYGEPRKL, amino acids residue and SAYGEPRKLIMNSRPPAV SAYGEPRKL
alanine substituted for amino acids 168-176 of MAGE-3 IMNSRPPAVEVDPIGHLY, cysteine at the eighth amino EVDPIGHLYIMNSRPPAV EVDPIGHLY
acid residue (IMNSRPPAV) amino acids 271-279 of MAGE-3 IMNSRPPAVFLWGPRALV, amino acids 167-176 of MAGE-3 IM1SRI'PAV1VIEVDPIGHLY, MEVDPIGHLYIMNSRPPAVMEVDPIGHLY
oe tµ.) tµ.) C35 Epitope Analog Exemplary Tumor Rejection Peptide Exemplary Polytope amino acids 2-10 of BAGE IMNSRPPAVAARAVFLAL, AARAVFLALIMNSRPPAVAARAVFLAL
amino acids 9-16 of GAGE-1,2 IMNSRPPAVYRPRPRRY, YRPRPRRYININSRPPAVYRPRPRRY
amino acids 11-20 of RAGE IMNSRPPAVSPSSNRIRNT, SP S SNRIRNTIMNSRPPAVSPS SNRIRNT
amino acids 23-32 of CDK4 IMNSRPPAVARDPHS GHFV, ARDPHSGHFVIMNSRPPAVARDPHS GHFV
amino acids 29-37 off3-catenin IMNSRPPAVSYLD S GIHS, SYLD S GIFISIMNSRPPAVSYLD SGIHS

amino acids 1-9 of Tyrosinase IMNSRPPAVMLLAVLYCL, MLLAVLYCLIMNSRPPAVMLLAVLYCL
oo 1,) amino acids 206-214 of Tyrosinase IMNSRPPAVAFLPWHRLF, AFLPWHRLFIMNSRPPAVAFLPWHRLF
amino acids 56-70 of Tyrosinase IMNSRPPAVQNILLSNAPLGPQFP, QNILLSNAPLGP QFPIMNSRPPAVQNILLSNAPLGPQFP
amino acids 448-462 of Tyrosinase IMNSRPPAVDYSYLQDSDPDSFQD, DYSYLQD SDPD SF QDIMNSRPPAVDYSYLQD SDPD SFQD
amino acids 32-40 of Melan-Am' IMNSRPPAVJLTVILGVL, JLTVILGVLIMNSRPPAVJLTVILGVL
amino acids 154-162 of gp100'11' IMNSRPPAVKTWGQYWQV, KIWGQYWQVIMNSRPPAVKTWGQYVVQV

amino acids 209-217 of gp10 OPme 1 17 IMNSRPPAVITDQVPFSV, ITDQVPFSVIMNSRPPAVITD QVPFSV
oe C35 Epitope Analog Exemplary Tumor Rejection Peptide Exemplary Polytope amino acids 280-288 of gp100P'117 IMNSRPPAVYLEPGPVTA, YLEPGPVTAIMNSRPPAVYLEPGPVTA
tµ.) oe amino acids 457-466 of gplOOP'117 IMNSRPPAVLLDGTATLRL, LLDGTATLRLIMNSRPPAVLLDGTATLRL
amino acids 476-485 of gplOOPT"117 IMNSRPPAVVLYRYGSFSV, VLYRYGSFSVIMNSRPPAVVLYRYGSFSV
amino acids 301-309 of PRAME IMNSRPPAVLYVDSLFFL, LYVDSLFFLIMNSRPPAVLYVDSLFFL
amino acids 292-303 of MAGE-6 IMNSRPPAVIUSGGPRISYPL, KISGGPRISYPLIMNSRPPAV'KISGGPRISYPL
co co amino acids 157-167 of NY-ESO-1 IMNSRPPAVSLLMWITQCFL, c7, co SLLMWITQCFLITNSRPPAVSLLMWITQCFL
oo amino acids 157-165 of NY-ESO-1 IMNSRPPAVSLLMWITQC, SLLMWITQCIMNSRPPAVSLLMWITQC

amino acids 155-163 of NY-ESO-1 IMNSRPPAVQLSLLMWIT, c7, QLSLLMWITIMNSRPPAVQLSLLMWIT
amino acids 157-170 of NY-ESO-1 SLLMWITQCFLPVFIMNSRPPAVTSYVKVLIIIIMVKISG
and amino acids 281-295 of MAGE-3 Analog of 1105 - V113 of amino acids 161-169 of MAGE-1 ITNSRPPSV EADPTGHSY, SEQ ID NO:2 having serine EADPTGHSYITNSRPPSVEADPTGHSY

substituted for cysteine at the eighth amino acid residue amino acids 230-238 of MAGE-1 ITNSRPPSV SAYGEPRKL, 1-3 (ITNSRPPSV) SAYGEPRKLITNSRPPSV SAYGEPRKL
amino acids 168-176 of MAGE-3 ITNSRPPSVEVDPIGHLY, oe EVDPIGHLYITNSRPPSV EVDPIGHLY
tµ.) tµ.) amino acids 271-279 of MAGE-3 ITNSRPPSVFLWGPRALV, FLWGPRALVITNSRPPSV FLWGPRALV
amino acids 167-176 of MAGE-3 ITNSRPPSVMEVDPIGHLY, MEVDPIGHLYITNSRPPSVMEVDPIGHLY
tµ.) oo amino acids 2-10 of BAGE ITNSRPP SVAARAVFLAL, AARAVFLALITNSRPP SVAARAVFLAL
amino acids 9-16 of GAGE-1,2 ITNSRPPSVYRPRPRRY, YRPRPRRYTTNSRPPSVYRPRPRRY
amino acids 11-20 of RAGE ITNSRPP SV SP S SNRIRNT, SP S SNRLRNTITNSRPPSVSPSSNRIRNT
amino acids 23-32 of CDK4 ITNSRPP SVARDPHS GHFV, ARDPHS GHFVITNSRPP SVARDPHSGHFV
co co c7, co amino acids 29-37 of P-catenin ITNSRPPSVSYLD S GUIS, oo "
SYLD S GIHSITNSRPPSVSYLD SGIHS

amino acids 1-9 of Tyrosinase ITNSRPPSVMLLAVLYCL, MLLAVLYCLITNSRPPSVMLLAVLYCL

c7, amino acids 206-214 of Tyrosinase ITNSRPPSVAFLPWHRLF, AFLPWHRLFITNSRPPSVAFLPWHRLF
amino acids 56-70 of Tyrosinase ITNSRPPSVQNILLSNAPLGPQFP, QNILLSNAPLGPQFPITNSRPPSVQNILLSNAPLGPQFP
amino acids 448-462 of Tyrosinase ITNSRPPSVDYSYLQDSDPDSFQD, DYSYLQDSDPDSFQDITNSRP_P SVDYSYLQD SDPDSFQD

amino acids 32-40 of Me1an-AmART4 ITNSRPPSVILTVILGVL, 1-3 JLTVILGVLITNSRPP SVJLTVILGVL
amino acids 154-162 of gp100117 ITNSRPPSVKTWGQYWQV, oo tµ.) KTWGQYVVQVITNSRPPSVKTWGQYWQV
tµ.) amino acids 154-162 of gp100'm GVRIVVEYAKTWGQYWQV, KTWGQYWQVGVRIVVEYAKTWGQYWQV
amino acids 209-217 of gp100Then7 GVRIVVEYAITDQVPFSV, IIDQVPFSVGVRIVVEYAITDQVPFSV
oe amino acids 280-288 of gp1001'117 GVRIVVEYAYLEPGPVTA, YLEPGPVTAGVRIVVEYAYLEPGPVTA
. amino acids 457-466 of gp10017 GVRIVVEYALLDGTATLRL, TinGTATLRLGVRIVVEYALLDGTATLRL
amino acids 476-485 of gp1001117 KLENGGFPVVLYRYGSFSV, VLYRYGSFSVKLENGGFPVVLYRYGSFSV
amino acids 301-309 of PRAME GVRIVVEYALYVDSLFFL, LYVDSLFFLGVRIVVEYALYVDSLFFL
(5) co amino acids 292-303 of MAGE-6 GVRIVVEYAKISGGPRISYPL, oo KISGGPRISYPLGVRIVVEYAKISGGPRISYPL

amino acids 157-167 of NY-ESO-1 GVRIVVEYASLLMWITQCFL, SLLMWITQCFLGVRIVVEYASLLMWITQC.FL

(5) amino acids 157-165 of NY-ES 0-1 GVRIVVEYASLLMWITQC, amino acids 155-163 of NY-ES0-1 GVRIVVEYAQL,SILMWIT, QLSLLMWIT GVRIVVEYAQLSLLMWIT
amino acids 157-170 of NY-ESO-land SLLMWITQCFLPVF GVRIVVEYATSYVKVLHEIMVKISG
amino acids 281-295 of MAGE-3 oe JI

amino acids 271-279 of MAGE-3 ICITNSRPPSVFLWGPRALV, FLWGPRALVKITNSRPPSV FLWGPRALV
amino acids 167-176 of MAGE-3 KITNSRPPSVMEVDPIGHLY, MEVDPIGHLYKITNSRPPSVMEVDPIGHLY
tµ.) oe amino acids 2-10 of BAGE KITNSRPPSVAARAVFLAL, amino acids 9-16 of GAGE-1,2 KITNSRPPSVYRPRPRRY, YRPRPRRYKITNSRPPSVYRPRPRRY
amino acids 11-20 of RAGE KITNSRPPSVSPSSNRJRNT, SPSSNRIRNTKITNSRPPSVSPSSNRIRNT

amino acids 23-32 of CDK4 KITNSRPPSVARDPHSGHFV, ARDPHSGHFVKITNSRPPSVARDPHSGHFV
co co c7, co amino acids 29-37 offi-catenin KITNSRPPSVSYLDSGIHS, oo SYLDSGIHSKITNSRPPSVSYLDSGIHS
"

amino acids 1-9 of Tyrosinase KITNSRPPSVMLLAVLYCL, amino acids 206-214 of Tyrosinase KITNSRPPSVAFLPWHRLF, AFLPWHRLFKITNSRPPSVAFLPWHRLF
amino acids 56-70 of Tyrosinase ICITNSRPPSVQNILLSNAPLGPQFP, QNILLSNAPLGPQFPKITNSRPPSVQNILLSNAPLGPQFP
amino acids 448-462 of Tyrosinase KITNSRPPSVDYSYLQDSDPDSFQD, DYSYLQDSDPDSFQDKITNSRPPSVDYSYLQDSDPDSFQD

amino acids 32-40 of Me1an-AmART4 ICITNSRPPSVJLTVILGVL, JLT'VILGVLICITNSRPPSVJLTVILGVL
amino acids 154-162 of gp100'117 KITNSRPPSVKTWGQYWQV, KTWGQYWQVKITNSRPPSVKTWGQYWQV
tµ.) amino acids 209-217 of gpl00Pme117 KITNSRPPSVITDQVPFSV, ITD QVPFSVKITNSRPPSVITD QVPFSV
amino acids 280-288 of gp100'117 KITNSRPPSVYLEPGPVTA, YLEPGPVTAKITNSRPPSVYLEPGPVTA
tµ.) oe amino acids 457-466 of gp 1 00P'117 KITNSRPPSVLLDGTATLRL, LLD GTATLRLKITNSRPP SVLLD GTATLRL
amino acids 476-485 of gpl00P"117 KITNSRPPSVVLYRYGSFSV, VLYRYGSFSVKITNSRPP SVVLYRYGSFSV
amino acids 301-309 of PRAME KITNSRPPSVLYVD SLFFL, LYVD SLFFLKITNSRPP SVLYVD SLFFL
amino acids 292-303 of MAGE-6 KITNSRPPSVKIS GGPRISYPL, 1.3 KIS GGPRISYPLKITNSRPPSVKIS GGPRISYPL
co co c7) amino acids 157-167 of NY-ESO-1 KITNSRPPSVSLLMWITQCFL, co SLLMWITQCFLKITNSRPPSVSLLM WITQCFL
1.3 amino acids 157-165 of NY-ESO-1 KITNSRPPSVSLLMWITQC, SLLMWITQCKITNSRPP SVSLLMWITQC
1.3 c7, amino acids 155-163 of NY-ESO-1 KITNSRPPSVQLSLLMWIT, QLSLLMWITKITNSRPP SVQLSLLMWIT
amino acids 157-170 of NY-ES 0-1 SLLIVIWITQCFLPVFICITNSRPPSVTSYVKVLHHMVKISG
and amino acids 281-295 of MAGE-3 Analog of G22 - C30 of SEQ amino acids 161-169 of MAGE-1 GVRIVVEYAEADPTGHSY, ID N0:2 having alanine EADPTGHSYGVRIVVEYAEADPTGHSY
substituted for cysteine at the ninth amino acid residue amino acids 230-238 of MAGE-1 GVRIVVEYASAYGEPRKL, 1-3 (GVRIVVEYA) SAYGEPRKLGVRIVVEYASAYGEPRKL
amino acids 168-176 of MAGE-3 GVRIVVEYAEVDPIGHLY, oe tµ.) EVDPIGHLYGVRIVVEYAEVDPIGHLY
tµ.) amino acids 271-279 of MAGE-3 GVRIVVEYAFLWGPRALV, FLWGPRALVGVRIVVEYAFLWGPRALV
amino acids 167-176 of MAGE-3 GVRIVVEYAMEVDPIGHLY, MEVDPIGHLYGVRIVVEYA.MEVDPIGHLY
oe amino acids 2-10 of BAGE GVRIVVEYAAARAVFLAL, AARAVFLALGVRIVVEYAAARAVFLAL
amino acids 9-16 of GAGE-1,2 GVRIVVEYAYRPRPRRY, YRPRPRRYGVRIVVEYAYRPRPRRY
amino acids 11-20 of RAGE GVRIVVEYASP S SNRIRNT, SP S SNRIRNTGVRIVVEYASPS SNRIRNT
amino acids 23-32 of CDK4 GVRIVVEYAARDPHSGHFV, co co ARDPHSGHFVGVRIVVEYAARDPHS GHFV
c7, co oo amino acids 29-37 of p-catenin GVRIVVEYASYLD SGIHS, SYLD SGIHS GVRIVVEYASYLD SGIHS
amino acids 1-9 of Tyrosinase GVRIVVEYAMLLAVLYCL, MLLAVLYCLGVRIVVEYAMLLAVLYCL
c7, amino acids 206-214 of Tyrosinase GVRIVVEYAAFLPWHRLF, AFLPWHRLFGVRIVVEYAAFLPWHRLF
amino acids 56-70 of Tyrosinase GVRIVVEYAQNILLSNAPLGPQFP, QNILLSNAPLGPQFP GVRIVVEYAQNILLSNAPLGPQFP
amino acids 448-462 of Tyrosinase GVRIVVEYADYSYLQDSDPDSFQD, S1-0 DYSYLQD SDPDSFQD GVRIVVEYADYSYLQD SDPD SF QD
amino acids 32-40 of Melan-Am'l GVRIVVEYAJLTVILGVL, JLTVILGVLGVRIVVEYAJLTVILGVL
oe amino acids 154-162 of gp100'117 GVRIVVEYAKTWGQYWQV, KTWGQYWQVGVRIVVEYAKTWGQYWQV
amino acids 209-217 of gplOOPmell7 GVRIVVEYAITDQVPFSV, ITD QVPFSVGVRIVVEYAITD QVPFSV
tµ.) oo amino acids 280-288 of gplOOPmel 17 GVRIVVEYAYLEPGPVTA, YLEPGPVTAGVRIVVEYAYLEPGPVTA
amino acids 457-466 of gplOOP'117 GVRIVVEYALLDGTATLRL, LLD GTATLRLGVRIVVEYALLD GTATLRL
amino acids 476-485 of gp100'117 KI,ENGGFPVVLYRYGSFSV, VLYRYGSFSVKLENGGFPVVLYRYGSFSV
amino acids 301-309 of PRAME
GVRIVVEYALYVDSLFFL, 0 LYVD SLFFLGVRIVVEYALYVD SLFFL
0.) amino acids 292-303 of MAGE-6 GVRIVVEYAKIS GGPRISYPL, KISGGPRISYPLGVRIVVEYAKISGGPRISYPL

amino acids 157-167 of NY-ESO-1 GVRIVVEYASLLMWITQCFL, SLLMWITQCFLGVRIVVEYASLLMWITQCFL

c7, amino acids 157-165 of NY-ESO-1 GVRIVVEYASLLMWITQC, amino acids 155-163 of NY-ES 0- 1 GVRIVVEYAQLSLLMWIT, QLSLLM WIT GVRIVVEYAQLSLLMWIT
amino acids 157-170 of NY-ESO-land SLLMWITQCFLPVF GVRIVVEYATSYVKVLHHMVICISG
amino acids 281-295 of MAGE-3 oe Analog of 125 - C33 of SEQ amino acids 161-169 of MAGE-1 IVVEYAEPAEADPTGHSY, ID NO:2 having alanine EADPTGHSYIVVEYAEPAEADPTGHSY
substituted for cysteine at the sixth and ninth amino acids amino acids 230-238 of MAGE-1 IVVEYAEPASAYGEPRKL, residues (IVVEYAEPA) SAYGEPRKLIVVEYAEPASAYGEPRKL
t=.) oo amino acids 168-176 of MAGE-3 IVVEYAEPAEVDPIGHLY, EVDPIGHLYIVVEYAEPAEVDPIGHLY
amino acids 271-279 of MAGE-3 1VVEYAEPAFLWGPRALV, FLWGPRALVIVVEYAEPAFLWGPRALV
amino acids 167-176 of MAGE-3 IVVEYAEPAMEVDPIGHLY, MEVDPIGHLYIVVEYAEPAMEVDPIGHLY
amino acids 2-10 of BAGE 1VVEYAEPAAARAVFLAL, co co c7, co amino acids 9-16 of GAGE-1,2 IVVEYAEPAYRPRPRRY, oo YRPRPRRYIVVEYAEPAYRPRPRRY

amino acids 11-20 of RAGE IVVEYAEPASPS SNIURNT, SP S SNRIRNTIVVEYAEPA SP S SNRIRNT

c7, amino acids 23-32 of CDK4 IVVEYAEPAARDPHS GHFV, ARDPHSGHFVIVVEYAEPAARDPHSGHFV
amino acids 29-37 of P-catenin IVVEYAEPASYLDSGIHS, SYLD S GIHSIVVEYAEPASYLDS GIHS
amino acids 1-9 of Tyrosinase 1VVEYAEPAMLLAVLYCL, MLLAVLYCLIVVEYAEPAMLLAVLYCL
amino acids 206-214 of Tyrosinase IVVEYAEPAAFLPWHRLF, AFLPWHRLFIVVEYAEPAAFLPWHRLF
oo JI
tµ.) tµ.) amino acids 56-70 of Tyrosinase IVVEYAEPAQNILLSNAPLGP QFP, QNILLSNAPLGPQFPIVVEYAEPAQNILLSNAPLGPQFP
amino acids 448-462 of Tyrosinase IVVEYAEPADYSYLQDSDPDSFQD, DYSYLQDSDPDSFQDIVVEYAEPADYSYLQDSDPDSFQD
tµ.) oe amino acids 32-40 of Me1an-AmART-1 IVVEYAEPAVJLTVILGVL, JLTVILGVLIVVEYAEPAJLTVILGVL
amino acids 154-162 of gplOOP'117 IVVEYAEPAKTWGQYWQV, KTWGQYWQVIVVEYAEPAKTWGQYWQV
amino acids 209-217 of gplOOP'117 IVVEYAEPAITDQVPFSV, ITDQVPFSVIVVEYAEPAITDQVPFSV

amino acids 280-288 of gp1001'17 IVVEYAEPAYLEPGPVTA, YLEPGPVTAIVVEYAEPAYLEP GPVTA
co co c7, co amino acids 457-466 of gp100'17 IVVEYAEPALLDGTATLRL, co LLD GTATLRLIVVEYAEPALLD GTATLRL
`P
amino acids 476-485 of gp100Pmel 17 IVVEYAEPAVLYRYGSFSV, VLYRYGSFSVIVVEYAEPAVLYRYGSFSV

c7, amino acids 301-309 of PRAME IVVEYAEPALYVDSLFFL, LYVDSLFFLIVVEYAEPALYVD SLFFL
amino acids 292-303 of MAGE-6 IVVEYAEPAKISGGPRISYPL, KIS GGPRIS YPLIVVEYAEPAKIS GGPRISYPL
amino acids 157-167 of NY-ESO-1 IVVEYAEPASLLMWITQCFL, SLLMWITQCFLIVVEYAEPASLLMWITQCFL

amino acids 157-165 of NY-ESO-1 IVVEYAEPASLLMWITQC, SLLMWITQCIVVEYAEPASLLMWITQC
amino acids 155-163 of NY-ESO-1 IVVEYAEPAQLSLLMWIT, oe tµ.) QLSLLMWIT IVVEYAEPAQLSLLM WIT

amino acids 157-170 of NY-ESO-land SLLMWITQCFLPVF IVVEYAEPATSYVKVLHHMVKISG
amino acids 281-295 of MAGE-3 Analog of K104 - C112 of amino acids 161-169 of MAGE-1 KITNSRPPAEADPTGHSY, SEQ ID Ng:2 having alanine EADPTGHSY KITNSRPPAEADPTGHSY
oe substituted for cysteine at the ninth amino acid residue amino acids 230-238 of MAGE-1 KITNSRPPASAYGEPRKL, (KITNSRPPA) S SAYGEPRKLKITNSRPPASAYGEPRICL
amino acids 168-176 of MAGE-3 K.ITNSRPPAEVDPIGHLY, EVDPIGHLYKITNSRPPAEVDPIGHLY
amino acids 271-279 of MAGE-3 KITNSRPPAFLWGPRALV, FLWGPRALVICITNSRPPAFLWGPRALV

amino acids 167-176 of MAGE-3 KITNSRPPAMEVDPIGHLY, MEVDPIGHLYKITNSRPPAMEVDPIGHLY
co co c7, co amino acids 2-10 of BAGE KITNSRPPAAARAVFLAL, AARAVFLALKITNSRPPAAARAVFLAL

amino acids 9-16 of GAGE-1,2 KITNSRPPAYRPRPRRY, YRPRPRRYKITNSRPPAYRPRPRRY

c7, amino acids 11-20 of RAGE KITNSRPPASP SSNRIRNT, SP S SNRIRNTKITNSRPPASP S SNRIRNT
amino acids 23-32 of CDK4 KITNSRPPAARDPHSGHFV, ARDPHSGHFVKITNSRPPAARDPHSGHFV
amino acids 29-37 of P-catenin KITNSRPPASYLDSGIHS, SYLDSGIHSKITNSRPPASYLDSGIHS

amino acids 1-9 of Tyrosinase KITNSRPPAMLLAVLYCL, MLLAVLYCLKITNSRPPAMLLAVLYCL
amino acids 206-214 of Tyrosinase KITNSRPPAAFLPWHRLF, tµ.) AFLPWHRLFK.ITNSRPPAAFLPWHRLF

amino acids 56-70 of Tyrosinase KITNSRPPAQNILLSNAPLGPQFP, QNILLSNAPLGP QFP KITNSRPPAQNILLSNAPLGP QFP
amino acids 448-462 of Tyrosinase KITNSRPPADYSYLQDSDPDSFQD, DYSYLQDSDPD SFQDICITNSRPPADYSYLQD SDPD SF QD
t=.) oe amino acids 32-40 of Me1an-AmART4 KITNSRPPAJLTVILGVL, JLTVILGVLKITNSRPPAYLTVILGVL
amino acids 154-162 of gplOOP'117 KITNSRPPAKTWGQYWQV, KTWGQYWQVKITNSRPPAKTWGQYWQV
amino acids 209-217 of gp100Pme117 KITNSRPPAITDQVPFSV, ITDQVPFSVKITNSRPPAITDQVPFSV

amino acids 280-288 of gp100' 17 KITNSRPPAYLEPGPVTA, YLEPGPVTAKITNSRPPAYLEP GPVTA
co co c7, CO
amino acids 457-466 of gpl V' K1TNSRPPALLDGTATLRL, LLD GTATLRLKITNSRPPALLD GTATLRL

amino acids 476-485 of gplOOP'117 KITNSRPPAVLYRYGSFSV, EL
VLYRYGSFSVKITNSRPPAVLYRYGSFSV

c7, amino acids 301-309 of PRAME KITNSRPPALYVDSLFFL, LYVDSLFFLKITNSRPPALYVDSLFFL
amino acids 292-303 of MAGE-6 KITNSRPPAKISGGPRISYPL, KISGGPRISYPLKITNSRPPAKIS GGPRISYPL
amino acids 157-167 of NY-ES 0-1 KITNSRPPASLLMWITQCFL, SLLMWITQCFLKITNSRPPASLLMWITQCFL

amino acids 157-165 of NY-ESO-1 KITNSRPPASLLMWITQC, SLLMWITQCKITNSRPPASLLMWITQC
amino acids 15-163 of NY-ESO-1 KITNSRPPAQLSLLMWIT, tµ.) QLSLLMWITKITNSRPPAQLSLLM WIT
tµ.) amino acids 157-170 of NY-ESO-1 SLLMWITQCFLPVFKITNSRPPATSYVKVLHHMVKISG
and amino acids 281-295 of MAGE-3 oe 1.) co co CO
=

JI
oe TABLED
C35 Epitope Exemplary Cell-Penetrating Exemplary Polypeptide Containing C35 Epitope and CPP
Peptide Sequence (CPP) S9 - V17 of' SEQ ID NO:2 HSV-1 tegument protein VP22 SVAPPPEEVMTSRRSVKSGPREVPRDEYEDLYYTPSSGMASPDSPPDTSRRGALQTRSRQRGEV
SVAPPPEEV RFVQYDESDYALYGGS S
SEDDEHPEVPRTRRPVS GAVL S GP GPARAP PPPA GS GGAGRTPTTA
PRAPRTQRVATKAPAAPAAETTRGRKSA QP ESAALPDAPA S TAP TRSKTPAQ GLARKLHF S TA
PPNPDAPWTPRVAGENKRVECAAVGRLAAMHARMAAVQLWDMSRPRTDEDLNELLGITTIR
VTVCEGKNLLQRANELVNPDVVQDVDAATATRGRSAASRPTERPRAPARSASRPRRPVE, MTSRRSVKSGPREVPRDEYEDLYYTPSSGMASPDSPPDTSRRGALQTRSRQRGEVRFVQYDES
DYALYGGSS SEDDEHPEVPRTRRPVS GAVLS GP GPARAP PPPA GS GGA GRTP TTAPRAPRTQR

VATKAPAAPAAETTRGRKSAQPESAALPDAPASTAPTRSKTPAQ GLARKLHF STAPPNPDAPW
co TPRVAGENKRVECAAVGRLAAMHARMAAVQLWDMSRPRIDEDLNELLGITTIRVTVCEGKN
co c7, LLQRANELVNPDVVQDVDAATATRGRSAASRPTERPRAPARSASRPRRPVESVAPPPEEVMTS
co RRSVKSGPREVPRDEYEDLYYTPSSGMASPD SPPDTSRRGALQTRSRQRGEVRFVQYDESDYA Lt"

LYGGSS SEDDEHPEVPRTRRPVS GAVLS GP GPARAPPPPAGS GGAGRTPTTAPRAPRTQRVATK

APAAPAAETTRGRKSAQPESAALPDAPASTAPTRSKTPAQ GLARKLHF S TAPPNPD APWTP RV
AGFNKRVECAAVGRLAAMHARMAAVQLWDMSRPRTDEDLNELLGITTIRVTVCEGKNLLQR

ANELVNPDVVQDVDAATATRGRSAASRPTERPRAPARSASRPRRPVE
c7, membrane-translocating SVAPPPEEVAAVLLPVLLAAP, sequence (MST) from h region AAVLLPVLLAAPSVAPPPEEVAAVLLPVLLAAP
of the signal sequence of Kaposi fibroblast growth factor oe C35 Epitope Exemplary Cell-Penetrating Exemplary Polypeptide Containing C35 Epitope and CPP
Peptide Sequence (CPP) S21 - Y29 of SEQ ID NO:2 HSV-1 tegument protein VP22 SGVRIVVEYMTSRRSVKSGPREVPRDEYEDLYYTPS S GMASPDSPPDTSRRGALQTRSRQRGE
SGVRIVVEY VRFVQYDESDYALYGGS S
SEDDEHPEVPRTRRPVS GAVLSGPGPARAPPPPAGSGGAGRTPTT
oe APRAPRTQRVATKAPAAPAAETTRGRKSAQPESAALPDAPASTAPTRSKTPAQGLARKLHFST
APPNPDAPWTPRVAGFNICRVECAAVGRLAAMHARMAAVQLWDMSRPRTDEDLNELLGITTI
RVTVCEGKNLLQRANELVNPDVVQDVDAATATRGRSAASRPTERPRAPARSASRPRRPVE, MTSRRSVKSGPREVPRDEYEDLYYTPSSGMASPDSPPDTSRRGALQTRSRQRGEVRFVQYDES
DYALYGGSSSEDDEHPEVPRTRRPVSGAVLSGPGPARAPPPPAGSGGAGRTPTTAPRAPRTQR
VATKAPAAPAAETTRGRKSAQPESAALPDAPAS TAPTRSKTPAQGLARKLHFSTAPPNPDAPW
TPRVAGENKRVFCAAVGRLAAMHARMAAVQLWDMSRPRTDEDLNELLGITTIRVTVCEGKN
LLQRANELVNPDVVQDVDAATATRGRSAASRPTERPRAPARSASRPRRPVESGVRIVVEYMTS

RRSVKSGPREVPRDEYEDLYYTP SSGMASPD SPPDTSRRGALQTRSRQRGEVRFVQYDESDYA
LYGGSSSEDDEHPEVPRTRRPVSGAVLSGPGPARAPPPPAGSGGAGRTPTTAPRAPRTQRVATK
co co APAAPAAEIER.GRKSAQPESAALPDAPASTAPTRSKTPAQGLARKLHFSTAPPNPDAPWTPRV
co AGFNKRVFCAAVGRLAAMHARMAAVQLWDMSRPRTDEDLNELLGITTIRVTVCEGKNLLQR
ANELVNPDVVQDVDAATATRGRSAASRPTERPRAPARSASRPRRPVE

=
membrane-translocating SGVRIVVEYAAVLLPVLLAAP, sequence (MST) from h region AAVLLPVLLAAPSGVRIVVEYAAVLLPVLLAAP

c7, of the signal sequence of Kaposi fibroblast growth factor oe JI
= w C35 Epitope Exemplary Cell-Penetrating Exemplary Polypeptide Containing C35 Epitope and CPP
Peptide Sequence (CPP) G22 - C30 of SEQ ID NO:2 HSV-1 tegument protein VP22 GVRIVVEYCMTSRRSVKSGPREVPRDEYEDLYYTPSSGMASPDSPPDTSRRGALQTRSRQRGE
GVRIVVEYC
VRFVQYDESDYALYGGSSSEDDEHPEVPRTRRPVSGAVLSGPGPARAPPPPAGSGGAGRTPTT
tµ.) oe APRAPRTQRVATKAPAAPAAE IIRGRKSAQPESAALPDAPASTAPTRSKTPAQGLARKLHFS T
APPNPDAPWTPRVAGFNKRVFCAAVGRLAAMHARMAAVQLWDMSRPRTDEDLNELLGITTI
RVTVCEGKNLLQRANELVNPDVVQDVDAATATRGRSAASRPTERPRAPARSASRPRRPVE, MTSRRSVKSGPREVPRDEYEDLYYTPSSGMASPDSPPDTSRRGALQTRSRQRGEVRFVQYDES
DYALYGGS S SEDDEHPEVPRTRRPVS GAVLS GP GPARAPPPPAGS GGAGRTPTTAPRAPRTQR
VATKAPAAPAAETTRGRKSAQPESAALPDAPASTAPTRSKTPAQGLARKLHFSTAPPNPDAPW
TPRVAGFNKRVFCAAVGRLAAMHARMAAVQLWDMSRPRTDEDLNELLGITTIRVTVCEGKN
LLQRANELVNPDVVQDVDAATATRGRSAASRPTERPRAPARSASRPRRPVEGVRIVVEYCMTS

RRSVKSGPREVPRDEYEDLYYTPSSGMASPDSPPDTSRRGALQ I'RSRQRGEVRFVQYDESDYA
LYGGSSSEDDEHPEVPRTRRPVSGAVLSGPGPARAPPPPAGSGGAGRTPTTAPRAPRTQRVATK
co co c7, APAAPAAE FIRGRKSAQPESAALPDAPASTAPTRSKTPAQGLARKLHFSTAPPNPDAPWTPRV
co AGFNKRVFCAAVGRLAAMHARMAAVQLWDMSRPRTDEDLNELLGITTIRVTVCEGKNLLQR
ANELVNPDVVQDVDAATATRGRSAASRPTERPRAPARSASRPRRPVE

membrane-translocating GVRIVVEYCAAVLLPVLLAAP, sequence (MST) from h region AAVLLPVLLAAPGVRIVVEYCAAVLLPVLLAAP

c7, of the signal sequence of Kaposi fibroblast growth factor = oe C35 Epitope Exemplary Cell-Penetrating Exemplary Polypeptide Containing C35 Epitope and CPP
Peptide Sequence (CPP) 125- C33 of SEQ ID NO:2 HSV-1 tegument protein VP22 IVVEYCEPCMTSRRSVKSGPREVPRDEYEDLYYTPSSGMASPDSPPDTSRRGALQTRSRQRGEV
IVVEYCEPC
RFVQYDESDYALYGGSSSEDDEHPEVPRTRRPVSGAVLS GP GPARAPPPPAGSGGAGRTPTTA
oe PRAPRTQRVATKAPAAPAAE ITRGRKSAQPESAALPDAPA S TAP TRSKTPA Q GLARKLHF STA
PPNPDAPWTPRVAGFNKRVFCAAVGRLAAMHARMAAVQLWDMSRPRTDEDLNELLGITTIR
VTVCEGKNLLQRANELVNPDVVQDVDAATATRGRSAASRPTERPRAPARSASRPRRPVE, MTSRRSVKSGPREVPRDEYEDLYYTP S SGMASPDSPPDTSKRGALQTRSRQRGEVRFVQYDES
DYALYGGSSSEDDEHPEVPRTRRPVSGAVLSGPGPARAPPPPAGSGGAGRTPTTAPRAPRTQR
VATKAPAAPAAETTRGRKSAQPESAALPDAPASTAPTRSKTPAQGLARKLHFSTAPPNPDAPW
TPRVAGFNKRVFCAAVGRLAAMHARMAAVQLWDMSRPRTDEDLNELLGITTIRVTVCEGKN
LLQRANELVNPDVVQDVDAATATRGRSAASRPTERPRAPARSASRPRRPVEIVVEYCEPCMTS

RRSVKSGPREVPRDEYEDLYYTP SSGMASPDSPPDTSRRGALQTRSRQRGEVRFVQYDESDYA
LYGGSSSEDDEHPEVPRTRRPVSGAVLSGPGPARAPPPPAGSGGAGRTPTTAPRAPRTQRVATK
c7, APAAPAAE fl".R.GRKSAQPESAALPDAPASTAPTRSKTPAQGLARKLHFSTAPPNPDAPWTPRV
AGFNICRVFCAAVGRLAAMHARMAAVQLWDMSRPRTDEDLNELLGITTIRVTVCEGKNLLQR
ANELVNPDVVQDVDAATATRGRSAASRPTERPRAPARSASRPRRPVE

membrane-translocating IVVEYCEPCAAVLLPVLLAAP, sequence (MST) from h region AAVLLPVLLAAPIVVEYCEPCAAVLLPVLLAAP

c7, of the signal sequence of Kaposi fibroblast growth factor oe C35 Epitope Exemplary Cell-Penetrating Exemplary Polypeptide Containing C35 Epitope and CPP
Peptide Sequence (CPP) T38 - V46 of SEQ ID NO:2 HSV-1 tegument protein VP22 TYLELASAVMTSRRSVKSGPREVPRDEYEDLYYTPSS GMASPDSPPDTSRRGALQTRSRQRGE
TYLELASAV
VRFVQYDESDYALYGGSSSEDDEHPEVPRTRRPVSGAVLSGPGPARAPPPPAGSGGAGRTPTT
tµ.) oe APRAPRTQRVATKAPAAPAAE1-1E.GRKSAQPESAALPDAPASTAPTRSKTPAQ GLARKLHF ST
APPNPDAPWTPRVAGFNKRVFCAAVGRLAAMHARMAAVQLWDMSRPRTDEDLNELLGITTI
RVTVCEGKNLLQRANELVNPDVVQDVDAATATRGRSAASRPTERPRAPARSASRPRRPVE, MTSRRSVKSGPREVPRDEYEDLYYTPSSGMASPDSPPDTSRRGALQTRSRQRGEVRFVQYDES
DYALYGGSSSEDDEHPEVPRTRRPVSGAVLSGPGPARAPPPPAGSGGAGRTPTTAPRAPRTQR
VATKAPAAPAAETTRGRKSAQPESAALPDAPASTAPTRSKTPAQGLARKLHFSTAPPNPDAPW
TPRVAGFNKRVFCAAVGRLAAMHARMAAVQLWDMSRPRTDEDLNELLGITTIRVTVCEGKN
LLQRANELVNPDVVQDVDAATA'TRGRSAASRPTERPRAPARSASRPRRPVETYLELASAVMTS

RRSVKSGPREVPRDEYEDLYYTPSSGMASPDSPPDTSRRGALQTRSRQRGEVRFVQYDESDYA
LYGGSSSEDDEHPEVPRTRRPVSGAVLSGPGPARAPPPPAGSGGAGRTPTTAPRAPRTQRVATK
co co APAAPAAETIRGRKSAQPESAALPDAPASTAPTRSKTPAQGLARKLHFSTAPPNPDAPWTPRV
c7, co AGFNKRVFCAAVGRLAAMHARMAAVQLWDMSRPRTDEDLNELLGITTIRVTVCEGKNLLQR
ANELVNPDVVQDVDAATATRGRSAASRPTERPRAPARSASRPRRPVE

membrane-translocating TYLELASAVAAVLLPVLLAAP, sequence (MST) from h region AAVLLPVLLAAP TYLELASAVAAVLLPVLLAAP

of the signal sequence of Kaposi c7, fibroblast growth factor oe JI

C35 Epitope Exemplary Cell-Penetrating Exemplary Polypeptide Containing C35 Epitope and CPP
Peptide Sequence (CPP) G61 -169 of SEQ ID NO:2 HSV-1 tegument protein VP22 GTGAFEIEIMTSRRSVKSGPREVPRDEYEDLYYTPS S GMASPD SPPDTSRRGALQTRSRQRGEV
GTGAFEIEI
RFVQYDESDYALYGGSSSEDDEHPEVPRTRRPVSGAVLSGPGPARAPPPPAGSGGAGRTPTTA
tµ.) oe PRAPRTQRVATKAPAAPAAETTRGRKSAQPESAALPDAPASTAPTRSKTPAQGLARKLHF STA
PPNPDAPWTPRVAGFNKRVFCAAVGRLAAMHARMAAVQLWDMSRPRTDEDLNELLGITTIR
VTVCEGKNLLQRANELVNPDVVQDVDAATATRGRSAASRPTERPRAPARSASRPRRPVE, MTSRRSVKSGPREVPRDEYEDLYYTPS SGMASPDSPPDTSRR GALQTRSRQRGEVRFVQYDES
DYALYGGSSSEDDEHPEVPRTRRPVSGAVLSGPGPARAPPPPAGSGGAGRTPTTAPRAPRTQR
VATKAPAAPAAETTRGRKSAQPESAALPDAPASTAPTRSKTPAQGLARKLHFSTAPPNPDAPW
TPRVAGFNKRVFCAAVGRLAAMHARMAAVQLWDMSRPRTDEDLNELLGITTIRVTVCEGKN
LLQRANELVNPDVVQDVDAATATRGRSAASRP IIRPRAPARSASRPRRPVEGTGAFEIEIMTSR

RSVKSGPREVPRDEYEDLYYTPSSGMASPDSPPDTSRRGALQTRSRQRGEVRFVQYDESDYAL
YGGS S SEDDEHPEVPRTRRPVSGAVLSGP GPARAPPPPAGSGGAGRTPTTAPRAPRTQRVATK
co co c7, co AGFNKRVFCAAVGRLAAMHARMAAVQLWDMSRPRTDEDLNELLGITTIRVTVCEGKNLLQR
ANELVNPDVVQDVDAATATRGRSAASRPTERPRAPARSASRPRRPVE

membrane-translocating GTGAFEIEIAAVLLPVLLAAP, sequence (MST) from h region AAVLLPVLLAAPGTGAFEIE1AAVLLPVLLAAP

c7, of the signal sequence of Kaposi fibroblast growth factor oe C35 Epitope Exemplary Cell-Penetrating Exemplary Polypeptide Containing C35 Epitope and CPP
Peptide Sequence (CPP) F65 - L73 of SEQ ID NO:2 HSV-1 tegument protein VP22 FEIEINGOLMTSRRSVKSGPREVPRDEYEDLYYTPSSGMASPDSPPDTSRRGALQTRSRQRGEV
FEIEINGQL RFVQYDESDYALYGGS S
SEDDEHPEVPRTRRPVSGAVLS GP GPARAPPPPA GS GGA GRTPTTA
oe PRAPRTQRVATKAPAAPAAE FIRGRKSAQPESAALPDAPAS TAP TRSKTPAQ GLARKLHF S TA
PPNPDAPWTPRVAGFNKRVFCAAVGRLAAMHARMAAVQLWDMSRPRTDEDLNELLGITTIR
VTVCEGKNLLQRANELVNPDVVQDVDAATATRGRSAASRPTERPRAPARSASRPRRPVE, MTSRRSVKS GP REVPRDEYEDLYYTP SSGMASPDSPPDTSRRGALQTRSRQRGEVRFVQYDES
DYALYGGS S SEDDEHPEVPRTRRPVS GAVLS GP GPARAPPPPA GS GGA GRTP TTAPRAPRTQR
VATKAPAAPAAETTRGRKSAQPESAALPDAPASTAPTRSKTPAQGLARKLHFSTAPPNPDAPW
TPRVAGFNKRVFCAAVGRLAAMHARMAAVQLWDMSRPRTDEDLNELLGITTIRVTVCEGKN
LLQRANELVNPDVVQDVDAATATRGRSAASRPTERPRAPARSASRPRRPVEFEIEINGOLMTSR

RSVKSGPREVPRDEYEDLYYTPS SGMASPD SPPDTSRRGALQTRSRQRGEVRFVQYD ES DYAL
YGGSS S ED D EHPEVPRTRRPVS GAVLS GP GPARAPPPPAGS GGAGRTPTTAPRAPRTQRVATK
co co c7, APAAPAAETTRGRKSAQPESAALPDAPASTAPTRSKTPAQGLARKLHFSTAPPNPDAPWTPRV Lco AGFNKRVFCAAVGRLAAMHARMAAVQLWDMSRPRTDEDLNELLGITTIRVTVCEGKNLLQR
1.) ANELVNPDVVQDVDAATATRGRSAASRPTERPRAPARSASRPRRPVE

membrane-translocating FEIEINGQLAAVLLPVLLAAP, sequence (MST) from h region AAVLLPVLLAAPFEIEENTGQLAAVLLPVLLAAP

c7, of the signal sequence of Kaposi fibroblast growth factor oe JI
=

C35 Epitope Exemplary Cell-Penetrating Exemplary Polypeptide Containing C35 Epitope and CPP
Peptide Sequence (CPP) 167 - F75 of SEQ ID NO:2 HSV-1 tegument protein VP22 lEINGQLVFMTSRRSVKSGPREVPRDEYEDLYYTPSSGMASPDSPPDTSRRGALQTRSRQRGEV
IEINGQLVF RFVQYDESDYALYGGS S
SEDDEHPEVPRTRRPVS GAVLS GP GPARAPPPPAGSGGAGRTPTTA tµ.) oe PRAPRTQRVATKAPAAPAAE l'IRGRICSAQPESAALPDAPASTAPTRSKTPAQ GLARICLHFS TA
PPNPDAPWTPRVAGFNKRVFCAAVGRLAAMHARMAAVQLWDMSRPRTDEDLNELLGITTIR
VTVCEGKNLLQRANELVNPDVVQDVDAATATRGRSAASRPTERPRAPARSASRPRRPVE, MTSRRSVKSGPREVPRDEYEDLYYTP SSGMASPD SPPDTSRRGALQTRSRQRGEVRFVQYDES
DYALYGGSSSEDDEHPEVPRTRRPVSGAVLS GP GPARAPPP PAGS GGAGRTP TTAPRAPRTQR
VATKAPAAPAAETTRGRKSAQPESAALPDAPASTAPTRSKTPAQGLARKLHFSTAPPNPDAPW
TPRVAGFNKRVFCAAVGRLAAMHARMAAVQLWDMSRPRTDEDLNELLGITTIRVTVCEGKN
LLQRANELVNPDVVQDVDAATATRGRSAASRPTERPRAPARSASRPRRPVEIEINGQLVFMTS

RRSVKSGPREVPRDEYEDLYYTP SSGMASPDSPPDTSRRGALQTRSRQRGEVRFVQYDESDYA
LYGGSSSEDDEHPEVPRTRRPVSGAVLSGPGPARAPPPPAGSGGAGRTPTTAPRAPRTQRVATK
c7, APAAPAAE FIRGRKSAQPESAALPDAPASTAPTRSKTPAQGLARKLHFSTAPPNPDAPWTPRV

ANELVNPDVVQDVDAATATRGRSAASRPTERPRAPARSASRPRRPVE

membrane-translocating IEINGOLVFAAVLLPVLLAAP, sequence (MST) from h region AAVLLPVLLAAPIEINGQLVFAAVLLPVLLAAP

c7, of the signal sequence of Kaposi fibroblast growth factor oe C35 Epitope Exemplary Cell-Penetrating Exemplary Polypeptide Containing C35 Epitope and CPP
Peptide Sequence (CPP) 1(77- Y85 of SEQ ID NO:2 HSV-1 tegument protein VP22 KLENGGFPYMTSRRSVKSGPREVPRDEYEDLYYTPS S GMASPDSPPDTSRRGALQTRSRQRGE
KLENGGFPY
VRFVQYDESDYALYGGSSSEDDEHPEVPRTRRPVSGAVLSGPGPARAPPPPAGSGGAGRTPTT
tµ.) oe APRAPRTQRVATKAPAAPAAETTRGRKSAQPESAALPDAPA S TAPTRSKTPAQGLARKLHFS T
APPNPDAPWTPRVAGENKRVECAAVGRLAAMHARMAAVQLWDMSRPRTDEDLNELLGITTI
RVTVCEGKNLLQRANELVNPDVVQDVDAATATRGRSAASRPTERPRAPARSASRPRRPVE, MTSRRSVKSGPREVPRDEYEDLYYTPSSGMASPDSPPDTSRRGALQTRSRQRGEVRFVQYDES
DYALYGGSSSEDDEHPEVPRTRRPVSGAVLSGPGPARAPPPPAGSGGAGRTPTTAPRAPRTQR
VATKAPAAPAAETTRGRKSAQPESAALPDAPASTAPTRSKTPAQGLARKLHESTAPPNPDAPW
TPRVAGENKRVECAAVGRLAAMHARMAAVQLWDMSRPRTDEDLNELLGITTIRVTVCEGKN
LLQRANELVNPDVVQDVDAATATRGRSAASRPTERPRAPARSASRPRRPVEKLENGGFYYMTS

RRSVKSGPRE'VPRDEYEDLYYTPSSGMASPDSPPDTSRRGALQTRSRQRGEVRFVQYDESDYA
LYGGSSSEDDEHPEVPRTRRPVSGAVLSGPGPARAPPPPAGSGGAGRTPTTAPRAPRTQRVATK
c7, APAAPAAETTRGRKSAQPESAALPDAPASTAPTRSKTPAQGLARKLEIFSTAPPNPDAPWTPRV
AGENKRVECAAVGRLAAMHARMAAVQLWDMSRPRIDEDLNELLGITTIRVTVCEGKNLLQR

ANELVNPDVVQDVDAATATRGRSAASRPTERPRAPARSASRPRRPVE

membrane-translocating KLENGGFPYAAVLLPVLLAAP, sequence (MST) from h region AAVLLPVLLAAPKLENGGFPYAAVLLPVLLAAP
of the signal sequence of Kaposi fibroblast growth factor oe C35 Epitope Exemplary Cell-Penetrating Exemplary Polypeptide Containing C35 Epitope and CPP
Peptide Sequence (CPP) Q72 - E86 of SEQ ID NO:2 HSV-1 tegument protein VP22 QLVFSKLENGGFPYEMTSRRSVKSGPREVPRDEYEDLYYTPSSGMASPDSPPDTSRRGALQTRS
QLVFSKLENGGFPYE
RQRGEVRFVQYDESDYALYGGSSSEDDEHPEVPRTRRPVSGAVLSGPGPARAPPPPAGSGGAG
oe RTPTTAPRAPRTQRVATKAPAAPAAE IlltGRKSAQPESAALPDAPASTAPTRSKTPAQGLARK
LHFSTAPPNPDAPWTPRVAGFNKRVFCAAVORLAAMHARMAAVQLWDMSRPRTDEDLNELL
Of MRVTVCEGKNLLQRANELVNPDVVQDVDAATATRGRSAASRPTERPRAPARSASRPRRPVE, MTSRRSVKSGPREVPRDEYEDLYYTPSSGMASPD SPPDTSRRGALQTRSRQRGEVRFVQYDES
DYALYGGSSSEDDEHPEVPRTRRPVSGAVLSGPGPARAPPPPAGSGGAGRTPTTAPRAPRTQR
VATKAPAAPAAETTRGRKSAQPESAALPDAPASTAPTRSKTPAQGLARKLHFSTAPPNPDAPW
TPRVAGFNKRVFCAAVORLAAMHARMAAVQLWDMSRPRTDEDLNELLGITTIRVTVCEGKN
LLQRANELVNPDVVQDVDAATATRGRSAASRPTERPRAPARSASRPRRPVEQLVFSKLENGGF

PYEMTSRRSVKSGPREVPRDEYEDLYYTPSSGMASPDSPPDTSRRGALQTRSRQRGEVRFVQY
DESDYALYGGSSSEDDEHPEVPRTRRPVSGAVLSGPGPARAPPPPAGSGGAGRTPTTAPRAPRT
co co QRVATKAPAAPAAETTRGRKSAQPESAALPDAPASTAPTRSKTPAQGLARKLHFSTAPPNPDA
c7, co PWTPRVAGFNKRVFCAAVORLAAMHARMAAVQLWDMSRPRTDEDLNELLGITTIRVTVCEG
KNLLQRANELVNPDVVQDVDAATATRGRSAASRPTERPRAPARSASRPRRPVE

membrane-translocating QLVFSKLENGGFPYEAAVLLPVLLAAP, sequence (MST) from h region AAVLLPVLLAAPQLVFSKLENGGFPYEAAVLLPVLLAAP

c7, of the signal sequence of Kaposi fibroblast growth factor oe C35 Epitope Exemplary Cell-Penetrating Exemplary Polypeptide Containing C35 Epitope and CPP
Peptide Sequence (CPP) G81 - L89 of SEQ ID NO:2 HSV-1 tegument protein VP22 GGFPYEKDLMTSRRSVKSGPREVPRDEYEDLYYTPS S GMASPDSPPDTSRRGALQTRSRQRGE
GGFPYEKDL VRFVQYDESDYALYGGS S
SEDDEHPEVPRTRRPVSGAVLS GP GPARAPPPPAGSGGAGRTPTT
tµ.) APRAPRTQRVATKAPAAPAAE'TTRGRKSAQPESAALPDAPASTAPTRSKTPAQGLARKLHF ST
oe APPNPDAPWTPRVAGFNKRVFCAAVGRLAAMHARMAAVQLWDMSRPRTDEDLNELLGITTI
RVTVCEGKNLLQRANELVNPDVVQDVDAATATRGRSAASRPTERPRAPARSASRPRRPVE, MTSRRSVKS GPREVPRDEYEDLYYTPSSGMASPD SPPDTSRRGALQTRSRQRGEVRFVQYDES
DYALYGGSSSEDDEHPEVPRTRRPVSGAVLSGPGPARAPPPPAGSGGAGRTPTTAPRAPRTQR
VATKAPAAPAAETTRGRKSAQPESAALPDAPASTAPTRSKTPAQGLARKLHFSTAPPNPDAPW
TPRVAGFNKRVFCAAVGRLAAMHARMAAVQLWDMSRPRTDEDLNELLGITTIRVTVCEGKN
LLQRANELVNPDVVQDVDAATATRGRSAASRPTERPRAPARSASRPRRPVEGGFPYEICDLMTS
RRSVKSGPREVPRDEYEDLYYTPSSGMASPDSPPDTSRRGALQTRSRQRGEVRFVQYDESDYA

LYGGSSSEDDEHPEVPRTRRPVSGAVLSGP GPARAPPPPAGSGGAGRTPITAPRAPRTQRVATK
co co APAAPAAETTRGRKSAQPESAALPDAPASTAPTRSKTPAQGLARKLHFSTAPPNPDAPWTPRV
co AGFNKRVFCAAVGRLAAMHARMAAVQLWDMSRPRTDEDLNELLGITTIRVTVCEGICNLLQR
ANELVNPDVVQDVDAATATRGRSAASRPTERPRAPARSASRPRRPVE

membrane-translocating GGFPYEKDLAAVLLPVLLAAP, sequence (MST) from h region AAVLLPVLLAAPGGFPYEKDLAAVLLPVLLAAP

of the signal sequence of Kaposi c7, fibroblast growth factor oe JI

C35 Epitope Exemplary Cell-Penetrating Exemplary Polypeptide Containing C35 Epitope and CPP
Peptide Sequence (CPP) K104 - C112 of SEQ ID HSV-1 tegument protein V222 KITNSRPPCMTSRRSVKSGPREVPRDEYEDLYYTPSSGMASPDSPPDTSRRGALQTRSRQRGEV
NO:2 RFVQYDESDYALYGGS S
SEDDEHPEVPRTRRPVS GAVLS GP GPARAPPPPAGS GGAGRTPTTA
tµ.) oe KITNSRPPC
PRAPRTQRVATKAPAAPAAETTRGRKSAQPESAALPDAPA STAPTRSKTPAQGLARKLHF STA
PPNPDAPWTPRVAGFNKRVFCAAVGRLAAMHARMAAVQLWDMSRPRTDEDLNELLGITTIR
VTVCEGKNLLQRANELVNPDVVQDVDAATATRGRSAASRP TERPRAPARSASRPRRPVE, MTSRRSVKS GPREVPRDEYEDLYYTP SS GMASPDSPPDTSRRGALQ'TRSRQRGEVRFVQYDES
DYALYGGSSSEDDEHPEVPRTRRPVS GAVLS GPGPARAPPPPAGS GGAGRTPTTAPRAPRTQR
VATKAPAAPAAETTRGRKSAQPESAALPDAPASTAPTRSKTPAQGLARKLHFSTAPPNPDAPW
TPRVAGFNKRVFCAAVGRLAAMHARMAAVQLWDMSRPRTDEDLNELLGITTIRVTVCEGKN
LLQRANELVNPDVVQDVDAATATRGRSAASRPTERPRAPARSASRPRRPVEKITNSRPPCMTS

RRSVKSGPREVPRDEYEDLYYTPSSGMASPDSPPDTSRRGALQTRSRQRGEVRFVQYDESDYA
LYGGS SSEDDEHPEVPRTRRPVS GAVLSGP GPARAPPPPAGS GGAGRTPTTAPRAPRTQRVATK =
co co APAAPAAETTRGRKSAQPESAALPDAPASTAPTRSKTPAQGLARKLHFSTAPPNPDAPWTPRV
c7, co AGFNKRVFCAAVGRLAAMHARMAAVQLWDMSRPRTDEDLNELLGITTIRVTVCEGKNLLQR 4c ANELVNPDVVQDVDAATATRGRSAASRPTERPRAPARSASRPRRPVE

membrane-translocating KITNSRPPCAAVLLPVLLAAP, sequence (MST) from h region AAVLLPVLLAAPKITNSRPPCAAVLLPVLLAAP

c7, of the signal sequence of Kaposi fibroblast growth factor oe C35 Epitope Exemplary Cell-Penetrating Exemplary Polypeptide Containing C35 Epitope and CPP
Peptide Sequence (CPP) K104 - V113 of SEQ ID HSV-1 tegument protein VP22 KITNSRPPCVMTSRRSVKSGPREVPRDEYEDLYYTPSSGMASPDSPPDTSRRGALQTRSRQRGE
NO:2 VRFVQYDESDYALYGGSSSEDDEHPEVPRTRRPVSGAVLSGPGPARAPPPPAGSGGAGRTPTT
KITNSRPPCV
APRAPRTQRVATKAPAAPAAET1RGRKSAQPESAALPDAPA STAPTRSKTPAQ GLARKLHF ST oe APPNPDAPWTPRVAGFNKRVFCAAVGRLAAMHARMAAVQLWDMSRPRTDEDLNELLGITTI
RVTVCEGKNLLQRANELVNPDVVQDVDAATATRGRSAASRPTERPRAPARSASRPRRPVE, MTSRRSVKSGPREVPRDEYEDLYYTPSSGMASPD SPPDTSRRGALQTRSRQRGEVRFVQYDES
DYALYGGSSSEDDEHPEVPRTRRPVSGAVLSGPGPARAPPPPAGSGGAGRTPTTAPRAPRTQR
VATICAPAAPAAETTRGRKSAQPESAALPDAPASTAPTRSKTPAQ GLARKLHFSTAPPNPDAPW
TPRVAGFNKRVFCAAVGRLAAMHARMAAVQLWDMSRPRTDEDLNELLGITTIRVTVCEGKN
LLQRANELVNPDVVQDVDAATATRGRSAASRPTERPRAPARSASRPRRPVEKITNSRPPCVMT

SRRSVKSGPREVPRDEYEDLYYTPSSGMASPDSPPDTSRRGALQTRSRQRGEVR_FVQYDESDY
ALYGGSSSEDDEHPE VPRTRRPVS GAVLSGP GPARAPPPPAGSGGAGRTPTTAPRAPRTQRVAT
co co KAPAAPAAETTRGRKSAQPESAALPDAPASTAPTRSKTPAQGLARKLHF STAPPNPDAPWTPR
c7, co VAGFNKRVFCAAVGRLAAMHARMAAVQLWDMSRPRTDEDLNELLGITTIRVTVCEGKNLLQ a, "
RANELVNPDVVQDVDAATATRQRSAASRPTERPRAPARSASRPRRPVE

membrane-translocating KITNSRPPCVAAVLLPVLLAAP, sequence (MST) from h region AAVLLPVLLAAPKITNSRPPCVAAVLLPVLLAAP
c7, of the signal sequence of Kaposi fibroblast growth factor oe C35 Epitope Exemplary Cell-Penetrating Exemplary Polypeptide Containing C35 Epitope and CPP
Peptide Sequence (CPP) 1105- V113 of SEQ ID NO:2 HSV-1 tegument protein VP22 ITNSRPPCVMTSRR.SVKSGPREVPRDEYEDLYYTPSSGMASPDSPPDTSRRGALQTRSRQRGEV
ITNSRPPCV
RFVQYDESDYALYGGSSSEDDEHPEVPRTRRPVSGAVLSGP GPARAPPPPAGSGGAGRTPTTA
tµ.) oe PRAPRTQRVATKAPAAPAAE l'IRGRKSAQPESAALPDAPASTAPTRSKTPAQGLARKLHFS TA
PPNPDAPWTPRVAGFNKRVFCAAVGRLAAMHARMAAVQLWDMSRPRTDEDLNELLGITTIR
VTVCEGKNLLQRANELVNPDVVQDVDAATAIRGRSAASRPTERPRAPARSASRPRRPVE, MTSRRSVKSGPREVPRDEYEDLYYTP S SGMASPD SPPDTSRRGALQTRSRQRGEVRFVQYDES
DYALYGGSSSEDDEHPEVPRTRRPVSGAVLSGPGPARAPPPPAGSGGAGRTPTTAPRAPRTQR
VATKAPAAPAAETTRGRKSAQPESAALPDAPASTAPTRSKTPAQGLARICLHFSTAPPNPDAPW
TPRVAGFNKRVFCAAVGRLAAMHARMAAVQLWDMSRPRTDEDLNELLG=RVTVCEGKN
LLQRANELVNPDVVQDVDAATATRGRSAASRPTERPRAPARSASRPRRPVEITNSRPPCVMTS

RRSVKSGPREVPRDEYEDLYYTP SSGMASPDSPPDTSRRGALQTRSRQRGEVRFVQYDESDYA
LYGGSSSEDDEHPEVPRTRRPVSGAVLSGPGPARAPPPPAGSGGAGRTPTTAPRAPRTQRVATK
co co APAAPAAETTRGRKSAQPESAALPDAPASTAPTRSKTPAQGLARKLHF STAPPNPDAPWTPRV
co AGFNKRVFCAAVGRLAAMHARMAAVQLWDMSRPRTDEDLNELLGITTIRVTVCEGKNLLQR
ANELVNPDVVQDVDAATATRGRSAASRPTERPRAPARSASRPRRPVE

membrane-translocating ITNSRPPCVAAVLLPVLLAAP, sequence (MST) from h region AAVLLPVLLAAPITNSRPPCVAAVLLPVLLAAP

c7, of the signal sequence of Kaposi fibroblast growth factor oe C35 Epitope Exemplary Cell-Penetrating Exemplary Polypeptide Containing C35 Epitope and CPP
Peptide Sequence (CPP) T101-V113 of SEQ ID NO:2 HSV-1 tegument protein VP22 TLEKITNSRPPCVMTSRRSVKSGPREVPRDEYEDLYYTPSS GMASPDSPPDTSRRGALQTRSRQ
TLEKITNSRPPCVIL RGEVRFVQYDESDYALYGGS S SEDDEHPEVPRTRRPVS GAVLS
GPGPARAPPPPAGS GGAGRT
tµ.) oe FSTAPPNPDAPWTPRVAGFNKRVFCAAVGRLAAMHARMAAVQLWDMSRPRTDEDLNELLGI
TTIRVTVCEGICNLLQRANELVNPDVVQDVDAATA fRGRSAASRPTERPRAPARSASRPRRPVE, MTSRRSVKS GPREVPRDEYEDLYYTP SS GMASPD SPPDTSRRGALQTRSRQRGEVRFVQYDES
DYALYGGSSSEDDEHPEVPRTRRPVS GAVLS GPGPARAPPPPAGS GGAGRTPTTAPRAPRTQR
VATKAPAAPAAETTRGRKSAQPESAALPDAPASTAPTRSKTPAQGLARKLHFSTAPPNPDAPW
TPRVAGFNKRVFCAAVGRLAAMHARMAAVQLWDMSRPRTDEDLNELLGITTIRVTVCEGKN
LLQRANELVNPDVVQDVDAATATRGRSAASRPTERPRAPARSASRPRRPVETLEKITNSRPPCV

MTSRRSVKSGPREVPRDEYEDLYYTPSS GMASPDSPPDTSRRGALQTRSRQRGEVRFVQYDES
DYALYGGSSSEDDEHPEVPRTRRPVSGAVLSGPGPARAPPPPAGS GGAGRTPTTAPRAPRTQR
co co VATKAPAAPAAETTRGRKSAQPESAALPDAPASTAPTRSKTPAQ GLARKLHFSTAPPNPDAPW
co TPRVAGFNKRVF CAAVGRLAAMHARMAAVQLWDMSRPRTDEDLNELLGITTIRVTVCEGKN F.) LLQRANELVNPDVVQDVDAATATRGRSAASRPTERPRAPARSASRPRRPVE

membrane-translocating TLEKITNSRPPCVAAVLLPVLLAAP, sequence (MST) from h region AAVLLPVLLAAPTLEKITNSRPPCVAAVLLPVLLAAP

c7, of the signal sequence of Kaposi fibroblast growth factor oe JI
= w C35 Epitope Exemplary Cell-Penetrating Exemplary Polypeptide Containing C35 Epitope and CPP
Peptide Sequence (CPP) 193 - V113 of SEQ ID NO:2 HSV-1 tegument protein VP22 IRRASNGETLEKITNSRPP LQTRSRQRGEVRFVQYDESDYALYGGS S S
ED D EHPEVPRTRRPVS GAVLS GP GPARAPPPPAG tµ.) oe CV
SGGAGRTPTTAPRAPRTQRVATKAPAAPAAETTRGRKSAQPESAALPDAPASTAPTRSKTPAQ
GLARKLHFSTAPPNPDAPWTPRVAGFNKRVFCAAVGRLAAMHARMAAVQLWDMSRPRTDE
DLNELLGITTIRVTVCEGKNLLQRANELVNPDVVQDVDAATATRGRSAASRPTERPRAPARSA
SRPRRPVE, MTSRRS VKS GPREVPRDEYEDLYYTP SS GMA SPD SPPDTSRRGALQTRSRQRGEVRFVQYDES
DYALYGGS S SEDDEHPEVPRTRRP VS GAVLS GP GPARAPPPPA GS GGA GRTP TTAP RAPRTQR
VATKA PAAPAAETTRGRKSAQPESAALPDAPASTAPTRSKTPAQGLARKLHF STAPPNPDAPW
TPRVAGFNKRVFCAAVGRLAAMHARMAAVQLWDMSRPRTDEDLNELLGITTIRVTVCEGKN

LLQRANELVNPDVVQDVDAATATRGRSAASRPTERPRAPARSA SRPRRP VEIRRA SNGETLEKI
co TNSRPP CVMTSRRSVKSGPREVPRDEYEDLYYTP SSGMASPDSPPDTSRRGALQTRSRQRGEV
co c7, RFVQYDESDYALYGGS S SEDDEHPEVPRTRRPVS GAVLS GP GPARAPPPPA GS GGA GRTPTTA
PRAPRTQRVATKAPAAPAAETTRGRKSAQPESAALPDAPASTAPTHSKTPAQGLARKLHFSTA F.c;

PPNPDAPWTPRVAGFNKRVFCAAVGRLAAMHARMAAVQLWDMSRPRTDEDLNELLGITTIR

VTVCEGKNLLQRANELVNPDVVQDVDAATATRGRSAASRPTERPRAPARSASRPRRPVE
membrane-trap slo eating lRRASNGETLEKITNSRPPCVAAVLLPVLLAAP, c7, sequence (MST) from h region AAVLLPVLLAAPIRRASNGETLE1CITNSRPPCVAAVLLPVLLAAP
of the signal sequence of Kaposi fibroblast growth factor oe JI

C35 Epitope Exemplary Cell-Penetrating Exemplary Polypeptide Containing C35 Epitope and CPP
Peptide Sequence (CPP) D88 - V113 of SEQ ID NO:2 HSV-1 tegument protein VP22 DLIEAIRRASNGETLEKITNSRPPCVMTSRRSVKSGPREVPRDEYEDLYYTPSSGMASPDSPPDT
DLIEAIRRASNGETLEKITN SRRGALQTRSRQRGEVRFV QYDESDYALYGGS
S SEDDEHPEVPRTRRPVS GAVLSGP GPARAP tµ.) oe SRPPCV PPPA GS GGA
GRTPTTAPRAPRTQRVATKAPAAPAAETTRGRKS AQPESAALPDAPA S TAPTRS
KTPAQ GLARKLHF STAPPNPDAPWTPRVAGFNKRVF CAAVGRLAAMHARMAAVQLWDMSR
PRTDEDLNELLGITTIRVTVCEGKNLLQRANELVNPDVVQDVDAATATRGRSAASRPTERPRA
PARSASRPRRPVE, MTSRRSVKSGPREVPRDEYEDLYYTPSSGMASPDSPPDTSRRGALQTRSRQRGEVRFVQYDES
D YALYGGS S SEDDEHPEVPRTRRPVS GAVLS GP GPARAPPPPA GS GGA GRTP TTAPRAPRTQR
VATKAPAAPAAETTRGRKSAQPESAALPDAPASTAPTRSKTPAQGLARKLHFSTAPPNPDAPW
TPRVAGFNKRVFCAAVGRLAAMHARMAAVQLWDMSRPRTDEDLNELLGITTIRVTVCEGKN

LLQRANELVNPDVVQDVDAATATRGRSAASRPTERPRAPARSASRPRRPVEDLIEAIRRASNGE
TLEKITNSRPP CVMTSRRSVKSGPREVPRDEYEDLYYTP SSGMASPD SPPDTSRRGALQTRSRQ
co co c7, RGEVRFVQYD E SD YALY G GS SSEDDEHPEVPRTRRPVS GAVLS GP GPARAPPPPA GS G GAGRT
co P TTAPRAPRTQRVATKAPAAPAAETTRGRKSA QPESAALPDAPAS TAP TRSKTPA Q GLARKLH
FSTAPPNPDAPWTPRVA GFNKRVF CAAVGRLAAMHARMAAVQLWDMSRPRTDEDLNELLGI

TTERVTVCEGKNLLQRANELVNPDVVQDVDAATATRGRSAASRPTERPRAPARSASRPRRPVE
membrane-translocating DLIEAIRRASNGETLEKITNSRPPCVAAVLLPVLLAAP, c7, sequence (MST) from h region AAVLLPVLLAAPDLIEAIRRASNGETLEKITNSRPPCVAAVLLPVLLAAP
of the signal sequence of Kaposi fibroblast growth factor oe C35 Epitope Exemplary Cell-Penetrating Exemplary Polypeptide Containing C35 Epitope and CPP
Peptide Sequence (CPP) P84 - V113 of SEQ ID NO:2 HSV-1 tegument protein VP22 PYEKDLIEAIRRASNGETLEKITNSRPPCVMTSRRSVKS GPREVPRDEYEDLYYTPSSGMASPDS
PYEKDLIEAIRRASNGETL
PPDTSRRGALQTRSRQRGEVRFVQYDESDYALYGGS S S EDD EHPEVPRTRRPVS GAVLS GP GP
tµ.) oe EKITNSRPPCV ARAPPPPAGSGGAGRTP
TTAPRAPRTQRVATKAPAAPAAETTRGRKSA QPESAALPDAPA S TA
PTRSKTPAQGLARKLHFSTAPPNPDAPWTPRVAGFNKRVFCAAVGRLAAMHARMAAVQLWD
MSRPRTDEDLNELLGITTIRVTVCEGKNLLQRANELVNPDVVQDVDAATATRGRSAASRPTER
PRAPARSASRPRRPVE, MTSRRSVKSGPREVPRDEYEDLYYTPSSGMASPDSPPDTSRRGALQTRSRQRGEVRFVQYDES
DYALYGGS S SEDDEHPEVPRTRRP VS GAVLS GP GPARAPPPPAGS GGAGRTPTTAPRAPRTQR
VATKAPAAPAAETTRGRKSAQPESAALPDAPASTAPTRSKTPAQGLARKLHFSTAPPNPDAPW
TPRVAGFNKRVF CAAVGRLAAMHARMAAVQLWDMSRPRTDEDLNELLGITTIRVTVCEGKN

LLQRANELVNPD VVQDVDAATATRGRSAASRPTERPRAPARSASRPRRPVEPYEKDLIEAIRRA
co SNGETLEKITNSRPPCVMTSRRSVKSGPREVPRDEYEDLYYTP SSGMASPD SPPDTSRRGALQT
co c7, RSRQRGEVRFVQYDESDYALYGGSSSEDDEHPEVPRTRRPVSGAVLSGPGPARAPPPPAGSGG
co AGRTP TTAPRAPRTQRVATKAPAAPAAEITRGRKSAQP ESAALPDAPA STAP TRSKTPA Q GLA

RICLHFSTAPPNPDAPWTPRVAGFNKRVECAAVGRLAAMHARMAAVQLWDMSRPRTDEDLN

ELLGITTIRVTVCEGKNLLQRANELVNPDVVQDVDAATATRGRSAASRPTERPRAPARSASRP
RRPVE

c7, membrane-translocating PYEKDLIEAIRRASNGETLEKITNSRPPCVAAVLLPVLLAAP, sequence (MST) from h region AAVLLPVLLAAPPYEKDLIEAIRRASNGETLEKITNSRPPCVAAVLLPVLLAAP
of the signal sequence of Kaposi fibroblast growth factor oe C35 Epitope Exemplary Cell-Penetrating Exemplary Polypeptide Containing C35 Epitope and CPP
Peptide Sequence (CPP) K77 -V113 of SEQ ID NO:2 HSV-1 tegument protein VP22 KLENGGFPYEKDLIEAERRASNGETLEKITNSRPPCVMTSRRSVKSGPREVPRDEYEDLYYTPSS
KLENGGFPYEKDLIEALRR.
GMASPDSPPDTSRRGALQTRSRQRGEVRFVQYDESDYALYGGSSSEDDEHPEVPRTRRPVSGA
ASNGETLEKITNSRPPCV VLS GPGPARAPPPPAGS
GGAGRTPTTAPRAPRTQRVATKAPAAPAAETTRGRKSAQPESAALP oe DAPASTAPTRSKTPAQGLARKLHFSTAPPNPDAPWTPRVAGFNKRVECAAVGRLAAMHARM
AAVQLWDMSRPRTDEDLNELLGITTIRVTVCEGKNLLQRANELVNPDVVQDVDAATATRGRS
AASRPTERPRAPARSASRPRRPVE, MTSRRSVKSGPREVPRDEYEDLYYTPS SGMASPDSPPDTSRRGALQTRSRQRGEVRFVQYDES
DYALYGGSSSEDDEHPEVPRTRRPVSGAVLSGPGPARAPPPPAGSGGAGRTPTTAPRAPRTQR
VATKAPAAPAAETTRGRKSAQPESAALPDAPASTAPTRSKTPAQGLARKLHFSTAPPNPDAPW
TPRVAGFNKRVFCAAVGRLAAMHARMAAVQLWDMSRPRTDEDLNELLGITTIRVTVCEGICN
LLQRANELVNPDVVQDVDAATATRGRSAASRPTERPRAPARSASRPRRPVEKLENGGFPYEKD

LIEAIRRASNGETLEKITNSRPPCVMTSRRSVKSGPREVPRDEYEDLYYTPSSGMASPDSPPDTS
RRGALQTRSRQRGEVRFVQYDESDYALYGGSSSEDDEHPEVPRTRRPVSGAVLSGPGPARAPP
PPAGSGGAGRTPTTAPRAPRTQRVATKAPAAPAAE FIRGRKSAQPESAALPDAPASTAPTRSK
TPAQGLARKLHFSTAPPNPDAPWTPRVAGFNKRVFCAAVGRLAAMHARMAAVQLWDMSRP

RTDEDLNELLGITTIRVTVCEGKNLLQRANELVNPDVVQDVDAATATRGRSAASRPTERPRAP
ARSASRPRRPVE

membrane-translocating KLENGGFPYEKDLIEAIRRASNGETLEKITNSRPPCVAAVELPVLLAAP, c7, sequence (MST) from h region AAVLLPVLLAAPKLENGGFPYEKDLIEAIRRASNGETLEKITNSRPPCV
AAVLLPVLLAAP
of the signal sequence of Kaposi fibroblast growth factor oe JI

C35 Epitope Exemplary Cell-Penetrating Exemplary Polypeptide Containing C35 Epitope and CPP
Peptide Sequence (CPP) Q72 - V113 of SEQ ID NO:2 HSV-1 tegument protein VP22 QLVFSKLENGGFPYEKDLIEAIRRASNGETLEKITNSRPPCVMTSRRSVKSGPREVPRDEYEDLY
QLVFSKLENGGFPYEKDLI
YTPSSGMASPDSPPDTSRRGALQTRSRQRGEVRFVQYDESDYALYGGSSSEDDEHPEVPRTRR
tµ.) oe EAIRRASNGETLEKITNSR
PVSGAVLSGPGPARAPPPPAGSGGAGRTPTTAPRAPRTQRVATKAPAAPAAETTRGRKSAQPE
PPCV
SAALPDAPASTAPTRSKTPAQGLARKLHFSTAPPNPDAPWTPRVAGFNKRVFCAAVGRLAAM
HARMAAVQLWDMSRPRTDEDLNELLGITTIRVTVCEGKNLLQRANEL VNPDVVQDVDAATA
TRGRSAASRPTERPRAPARSASRPRRPVE, MTSRRSVKSGPREVPRDEYEDLYYTPSSGMASPDSPPDTSRRGALQTRSRQRGEVRFVQYDES
DYALYGGSSSEDDEHPEVPRTRRPVSGAVLSGPGPARAPPPPAGS GGAGRTPTTAPRAPRTQR
VATKAPAAPAAETTRGRKSAQPESAALPDAPASTAP IRSKTPAQGLARICLHFSTAPPNPDAPW
TPRVAGFNKRVFCAAVGRLAAMHARMAAVQLWDMSRPRTDEDLNELLGITTIRVTVCEGKN

LLQRANELVNPDVVQDVDAATATRGRSAASRPTERPRAPARSASRPRRPVEQLVFSKLENGGF
PYEKDLIEAIRRASNGETLEICITNSRPPCVMTSRRSVKSGPREVPRDEYEDLYYTPSSGMASPDS
co co c7, PPDTSRRGALQTRSRQRGEVRFVQYDESDYALYGGSSSEDDEHPEVPRTRRPVSGAVLSGPGP
co ARAPPPPAGSGGAGRTPTTAPRAPRTQRVATKAPAAPAAETTRGRKSAQPESAALPDAPASTA

MSRPRTDEDLNELLGITTIRVTVCEGKNLLQRANELVNPDVVQDVDAATATRGRSAASRPTER
PRAPARSASRPRRPVE

c7, membrane-translocating OLVFSICLENGGFPYEKDLIEAIRRASNGETLEKITNSRPPCVAAVLLPVLLAAP, sequence (MST) from h region AAVLLPVLLAAPQLVFSKLENGGFPYEKDLIEAIRRASNGETLEKITNSRPPCVAAVLLPVLLAAP
of the signal sequence of Kaposi fibroblast growth factor oe C35 Epitope Exemplary Cell-Penetrating Exemplary Polypeptide Containing C35 Epitope and CPP

Peptide Sequence (CPP) o 1¨, F65 - V113 of SEQ ID NO:2 HSV-1 tegument protein VP22 FEIEINGOLVFSKLENGGFPYEKDLIEAIRRASNGETLEKITNSRPPCVMTSRRSVKSGPREVPRD
.6.
FEIEINGQLVFSKLENGGF EYEDLYYTPS SGMASPD
SPPDTSRRGALQTRSRQRGEVRFVQYDESDYALYGGSSSEDDEHPE .6.
tµ.) oe PYEKDLIEAIRRASNGETL
VPRTRRPVSGAVLSGPGPARAPPPPAGSGGAGRTPTTAPRAPRTQRVATKAPAAPAAETTRGR
EKITNSRPPCV
KSAQPESAALPDAPASTAPTRSKTPAQGLARKLHFSTAPPNPDAPWTPRVAGFNKRVFCAAVG
RLAAMHARMAAVQLWDMSRPRTDEDLNELLGITTIRVTVCEGKNLLQRANELVNPDVVQDV
DAATATRGRSAASRPTERPRAPARSASRPRRPVE, MTSRRSVKSGPREVPRDEYEDLYYTPSSGMASPDSPPDTSRRGALQTRSRQRGEVRFVQYDES
DYALYGGSSSEDDEHPEVPRTRRPVSGAVLSGPGPARAPPPPAGSGGAGRTPTTAPRAPRTQR
VATKAPAAPAAETTRGRKSAQPESAALPDAPASTAPTRSKTPAQGLARKLHFSTAPPNPDAPW
n TPRVAGFNKRVFCAAVGRLAAMHARMAAVQLWDMSRPRTDEDLNELLGITTIRVTVCEGKN

LLQRANELVNPDVVQDVDAATATRGRSAASRPTERPRAPARSASRPRRPVEFEIEINGQLVFSK
"
a, LENGGFPYEKDLIEAIRRASNGETLEKITNSRPPCVMTSRRSVKSGPREVPRDEYEDLYYTPSSG
co co MASPDSPPDTSRRGALQTRSRQRGEVRFVQYDESDYALYGGSSSEDDEHPEVPRTRRPVSGAV
co "
LSGPGPARAPPPPAGSGGAGRTPTTAPRAPRTQRVATKAPAAPAAETTRGRKSAQPESAALPD
APASTAPTRSKTPAQGLARKLHFSTAPPNPDAPWTPRVAGFNKRVFCAAVGRLAAMHARMA I

AVQLWDMSRPRTDEDLNELLGITTIRVTVCEGKNLLQRANELVNPDVVQDVDAATA'TRGRSA
a, H
ASRPTERPRAPARSASRPRRPVE
I.) c7, membrane-translocating FETEINGOLVFSKLENGGFPYEKDLIEAIRRASNGETLEKITNSRPPCVAAVLLPVLLAAP, sequence (MST) from h region AAVLLPVLLAAPFEIEINGOLVFSKLENGGFPYEKDLIEAIRRASNGETLEKITNSRPPCVAAVLL
of the signal sequence of Kaposi PVLLAAP
fibroblast growth factor Iv n ,-i cp =
oe w u, w C35 Epitope Exemplary Cell-Penetrating Exemplary Polypeptide Containing C35 Epitope and CPP
Peptide Sequence (CPP) L59 -V113 of SEQ ID NO:2 HSV-1 tegument protein VP22 LGGTGAFEIEINGQLVFSK REVPRDEYEDLYYTP

tµ.) oe LENGGFPYEKDLIEAIRRA DDEHPEVPRTRRP VS GAVLS GP
GPARAPPPPA GS GGA GRTP TTAPRAPRTQRVATKAPAAPAA
SNGETLEKITNSRPPCVIL
ETTRGRKSAQPESAALPDAPASTAPTRSKTPAQGLARKLHF STAPPNPDAPWTPRVAGFNKRV
FCAAVGRLAAMHARMAAVQLWDMSRPRTDEDLNELLGITTIRVTVCEGKNLLQRANELVNP
DVVQDVDAATATRGRSAASRPTERPRAPARSASRPRRPVE, MTSRRSVKSGPREVPRDEYEDLYYTPSSGMASPDSPPDTSRRGALQTRSRQRGEVRFVQYDES
DYALYGGS S SEDDEHP EVPRTRRPVS GAVLS GP GPARAPPPPA GS GGAGRTP TTAPRAP RTQR
VATKAPAAPAAETTRGRKSAQPESAALPDAPASTAP IRSKTPAQGLARKLHFSTAPPNPDAPW
TPRVAGFNKRVFCAAVGRLAAMHARMAAVQLWDMSRPRTDEDLNELLGITTIRVTVCEGKN

LLQRANELVNPDVVQDVDAATATRGRSAASRPTERPRAPARSASRPRRPVELGGTGAFEIEING
QLVFSKLENGGFPYEKDLIEAIRRASNGETLEKITNSRPPCVMTSRRSVKSGPREVPRDEYEDLY
co co YTPSSGMASPDSPPDTSRRGALQTRSRQRGEVRFVQYDESDYALYGGSSSEDDEHPEVPRTRR
c7, co PVSGAVLSGPGPARAPPPPAGSGGAGRTPTTAPRAPRTQRVATKAPAAPAAETTRGRKSAQPE

HARMAAVQLWDMSRPRTDEDLNELLGITTIRVTVCEGKNLLQRANELVNPDVVQDVDAATA
TRGRSAASRPTERPRAPARSASRPRRPVE

c7, membrane-translocating LGGTGAFEIEINGQLVFSKLENGGFPYEICDLIEAIRRASNGETLEKITNSRPPCVAAVLLPVLLAAP, sequence (MST) from h region AAVLLPVLLAAPLGGTGAFEIEINGOLVFSKLENGGFPYEKDLIEAIRRASNGETLEKITNSRPPCV
of the signal sequence of Kaposi AAVLLPVLLAAP
fibroblast growth factor oe JI

G99 - V113 of SEQ ID NO:2 HSV-1 tegument protein VP22 GETLEKITNSRPPCV_MTSRRSVKSGPREVPRDEYEDLYYTPSSGMASPDSPPDTSRRGALQTRS
GETLEKITNSRPPCV
RQRGEVRFVQYDESDYALYGGSSSEDDEHPEVPR'TRRPVSGAVLSGPGPARAPPPPAGSGGAG
RTPTTAPRAPRTQRVATKAPAAPAAETTRGRKSAQPESAALPDAPASTAPTRSKTPAQGLARK
= o LHFSTAPPNPDAPWTPRVAGFNKRVFCAAVGRLAAMHARMAAVQLWDMSRPRTDEDLNELL
GILLIRVTVCEGKNLLQRANELVNPDVVQDVDAATATRGRSAASRPTERPRAPARSASRPRRPVE, tµ.) oe MTSRRSVKSGPREVPRDEYEDLYYTPSSGMASPDSPPDTSRRGALQTRSRQRGEVRFVQYDES
DYALYGGSSSEDDEHPEVPRTRRPVSGAVLSGPGPARAPPPPAGSGGAGRTPTTAPRAPRTQR
VATKAPAAPAAETTRGRKSAQPESAALPDAPASTAPTRSKTPAQGLARKLHFSTAPPNPDAPW
TPRVAGFNKRVFCAAVGRLAAMHARMAAVQLWDMSRPRTDEDLNELLGITTIRVTVCEGKN
LLQRANELVNPDVVQDVDAATATRGRSAASRPTERPRAPARSASRPRRPVEGETLEKITNSRPP
CVMTSRRSVKSGPREVPRDEYEDLYYTPSSGMASPD SPPDTSRRGALQTRSRQRGEVRFVQYD
ESDYALYGGSSSEDDEHPEVPRTRRPVSGAVLSGPGPARAPPPPAGSGGAGRTPTTAPRAPRTQ
RVATKAPAAPAAETTRGRKSAQPESAALPDAPASTAPTRSKTPAQGLARKLHFSTAPPNPDAP

WTPRVAGFNKRVFCAAVGRLAAMHARMAAVQLWDMSRPRTDEDLNELLGITTIRVTVCEGK
co NLLQRANELVNPDVVQDVDAATATRGRSAASRPTERPRAPARSASRPRRPVE
co c7, membrane-translocating GETLEKITNSRPPCVAAVLLPVLLAAP, t71 sequence (MST) from h region AAVLLPVLLAAPGETLEKITNSRPPCVAAVLLPVLLAAP 0 of the signal sequence of Kaposi fibroblast growth factor c7, oe E100 - V113 of SEQ ID NO:2 HSV-1 tegument protein VP22 ETLEKITNSRPPCVMTSRRSVKSGPREVPRDEYEDLYYTPSSGMASPD SPPDTSRRGALQTRSR
ETLEKITNSRPPCV
QRGEVRFVQYDESDYALYGGSSSEDDEHPEVPRTRRPVSGAVLSGPGPARAPPPPAGSGGAGR
TPTTAPRAPRTQRVATKAPAAPAAETTRGRKSAQPESAALPDAPASTAPTRSKTPAQGLARKL
HFSTAPPNPDAPWTPRVAGFNKRVFCAAVGRLAAMHARMAAVQLWDMSRPRTDEDLNELL
GITTIRVTVCEGKNLLQRANELVNPDVVQDVDAATATRGRSAASRPTERPRAPARSASRPRRPVE, tµ.) oe MTSRRSVKSGPREVPRDEYEDEYYTPSSGMASPDSPPDTSRRGALQTRSRQRGEVRFVQYDES
DYALYGGSSSEDDEHPEVPRTRRPVSGAVLSGPGPARAPPPPAGSGGAGRTPTTAPRAPRTQR
VATKAPAAPAAETTRGRKSAQPESAALPDAPASTAPTRSKTPAQGLARKLHFSTAPPNPDAPW
TPRVAGFNKRVFCAAVGRLAAMHARMAAVQLWDMSRPRIDEDLNELLGITTIRVTVCEGKN ' LLQRANELVNPDVVQDVDAATATRGRSAASRPTERPRAPARSASRPRRPVEETLEKITNSRPPC
VMTSRRSVKSGPREVPRDEYEDLYYTP SSGMASPDSPPDTSRRGALQTRSRQRGEVRFVQYDE
SDYALYGGSSSEDDEHPEVPRTRRPVSGAVLSGPGPARAPPPPAGSGGAGRTPTTAPRAPRTQR
VATKAPAAPAAETTRGRKSAQPESAALPDAPASTAPTRSKTPAQGLARKLHFSTAPPNPDAPW

TPRVAGFNKRVFCAAVGRLAAMHARMAAVQLWDMSRPRTDEDLNELLGITTIRVTVCEGKN
LLQRANELVNPDVVQDVDAATATRGRSAASRPTERPRAPARSASRPRRPVE
co co c7, co membrane-translocating ETLEKITNSRPPCVAAVLLPVLLAAP, sequence (MST) from h region AAVLLPVLLAAPETLEKITNSRPPCVAAVLLPVLLAAP

of the signal sequence of Kaposi fibroblast growth factor 1:71 JI
oe TABLE E
C35 Epitope Analog Exemplary Cell-Penetrating Exemplary Polypeptide Containing C35 Epitope Analog and CPP
tµ.) oe Peptide Sequence (CPP) Analog of S77 -Y85 of SEQ HSV-1 tegument protein VP22 KLENGGFPVMTSRRSVKSGPREVPRDEYEDLYYTPSSGMASPDSPPDTSRRGALQTRSRQRGE
ID NO:2 having value VRFVQYDESDYALYGGSSSEDDEHPEVPRTRRPVSGAVLSGPGPARAPPPPAGSGGAGRTPTT
substituted for tyrosine at APRAPRTQRVATICAPAAPAAETTRGRKSAQPESAALPDAPASTAPTRSKTPAQGLARKLHFST
ninth amino acid residue APPNPDAPWTPRVAGFNKRVFCAAVGRLAAMHARMAAVQLWDMSRPRTDEDLNELLGITTI
(KLENGGFPV) RVTVCEGKNLLQRANELVNPDVVQDVDAATATRGRSAASRPTERPRAPARSASRPRRPVE, MTSRRSVKSGPREVPRDEYEDLYYTPSSGMASPDSPPDTSRRGALQTRSRQRGEVRFVQYDES
DYALYGGSSSEDDEHPEVPRTRRPVSGAVLSGPGPARAPPPPAGSGGAGRTPTTAPRAPRTQR

VATKAPAAPAAETTRGRKSAQPESAALPDAPASTAPTRSKTPAQGLARKLHFSTAPPNPDAPW
co co TPRVAGFNKRVFCAAVGRLAAMHARMAAVQLWDMSRPRTDEDLNELLGITTIRVTVCEGKN c7, co LLQRANELVNPDVVQDVDAATATRGRSAASRPTERPRAPARSASRPRRPVEKLENGGFPVMTS
RRSVKSGPREVPRDEYEDLYYTPSSGMASPDSPPDTSRRGALQTRSRQRGEVRFVQYDESDYA

LYGGSSSEDDEHPEVPRTRRPVSGAVLSGPGPARAPPPPAGSGGAGRTPTTAPRAPRTQRVATK
APAAPAAETTRGRKSAQPESAALPDAPASTAPTRSKTPAQGLARKLHFSTAPPNPDAPWTPRV
AGFNKRVFCAAVGRLAAMHARMAAVQLWDMSRPRTDEDLNELLGITTIRVTVCEGKNLLQR

ANELVNPDVVQDVDAATATRGRSAASRPTERPRAPARSASRPRRPVE
c7, membrane-translocating KLENGGFPVAAVLLPVLLAAP, sequence (MST) from h region AAVLLPVLLAAPKLENGGFPVAAVLLPVLLAAP
of the signal sequence of Kaposi fibroblast growth factor oe JI

C35 Epitope Analog Exemplary Cell-Penetrating Exemplary Polypeptide Containing C35 Epitope Analog and CPP
Peptide Sequence (CPP) Analog of K104 - C112 of HSV-1 tegument protein VP22 ICITNSRPPLMTSRRSVKSGPREVPRDEYEDLYYTPSSGMASPDSPPDTSRRGALQTRSRQRGEV
SEQ ID NO:2 having leucine RFVQYDESDYALYGGS
SSEDDEHPEVPRTRRPVS GAVLSGPGPARAPPPPAGSGGAGRTPTTA
oe substituted for cysteine at the PRAPRTQRVATKAPAAPAAETERGRICSAQPESAALPDAPAS TAPTRSKTPAQGLARKLHFS TA
ninth amino acid residue PPNPDAPWTPRVAGFNKRVFCAAVGRLAAMHARMAAVQLWDMSRPRTDEDLNELLGITTIR
(KITNSRPPL) VTVCEGKNLLQRANELVNPDVVQDVDAATATRGRSAASRPTERPRAPARSASRPRRPVE, MTSRRSVKSGPREVPRDEYEDLYYTP SSGMA SPD SPPDTSRRGALQTRSRQRGEVRFVQYDES
DYALYGGSSSEDDEHPEVPRTRRPVSGAVLSGPGPARAPPPPAGSGGAGRTPTTAPRAPRTQR
VATKAPAAPAAETTRGRKSAQPESAALPDAPA STAPTRSKTPAQGLARKLHF STAPPNPDAPW
TPRVAGFNKRVFCAAVGRLAAMHARMAAVQLWDMSRPRTDEDLNELLGITTIRVTVCEGKN
LLQRANELVNPDVVQDVDAATATRGRSAASRPTERPRAPARSA SRPRRPVEKITNSRPPLMTS

LYGGSSSEDDEHPEVPRTRRPVS GAVLSGP GPARAPPPPAGSGGAGRTPTTAPRAPRTQRVATK
co co APAAPAAETTRGRKSAQPESAALPDAPASTAPTRSKTPAQGLARICLHFSTAPPNPDAPWTPRV
co ANELVNPDVVQDVDAATATRGRSAASRPTERPRAPARSASRPRRPVE

membrane-translocating ICITNSRPPLAAVLLPVLLAAP, sequence (MST) from h region AAVLLPVLLAAPKITNSRPPLAAVLLPVLLAAP

(5) of the signal sequence of Kaposi fibroblast growth factor oe JI

C35 Epitope Analog Exemplary Cell-Penetrating Exemplary Polypeptide Containing C35 Epitope Analog and CPP
Peptide Sequence (CPP) Analog of 1105 - V113 of HSV-1 tegument protein VP22 ILNSRPPAVMTSRRSVKSGPREVPRDEYEDLYYTPSSGMASPDSPPDTSRRGALQTRSRQRGEV
SEQ ID NO:2 having leucine RFVQYDESDYALYGGSSSEDDEHPEVPRTRRPVSGAVLSGPGPARAPPPPAGSGGAGRTPTTA
tµ.) oe substituted for threonine at the PRAPRTQRVATKAPAAPAAETTRGRKSAQPESAALPDAPASTAPTRSKTPAQGLARKLHF STA
second amino acid residue PPNPDAPWTPRVAGFNKRVFCAAVGRLAAMHARMAAVQLWDMSRPRTDEDLNELLGITTIR
and alanine substituted for VTVCEGKNLLQRANELVNPDVVQDVDAATATRGRSAASRPTERPRAPARSASRPRRPVE, cysteine at the eighth amino acid residue (ILNSRPPCV) MTSRRSVKS GPREVPRDEYEDLYYTP S S
GMASPDSPPDTSRRGALQTRSRQRGEVRFVQYDES
DYALYGGSSSEDDEHPEVPRTRRPVSGAVLSGPGPARAPPPPAGSGGAGRTPTTAPRAPRTQR
VATKAPAAPAAETTRGRKSAQPESAALPDAPASTAPTRSKTPAQGLARKLHFSTAPPNPDAPW
TPRVAGFNKRVFCAAVGRLAAMHARMAAVQLWDMSRPRTDEDLNELLGITTIRVTVCEGKN
LLQRANELVNPDVVQDVDAATATRGRSAASRPTERPRAPARSASRPRRPVEILNSRPPAVMTS

RRSVKSGPREVPRDEYEDLYYTP SSGMASPDSPPDTSRRGALQTRSRQRGEVRFVQYDESDYA
LYGGSSSEDDEHPEVPRTRRPVSGAVLSGPGPARAPPPPAGSGGAGRTPTTAPRAPRTQRVATK
co co c7, APAAPAAELURGRKSAQPESAALPDAPASTAPTRSKTPAQGLARKLHFSTAPPNPDAPWTPRV
AGFNKRVFCAAVGRLAAMHARMAAVQLWDMSRPRTDEDLNELLGITTIRVTVCEGKNLLQR
ANELVNPDVVQDVDAATATRGRSAASRP fERPRAPARSASRPRRPVE

membrane-translocating ILNSRPPAVAAVLLPVLLAAP, sequence (MST) from h region AAVLLPVLLAAPILNSRPPAVAAVLLPVLLAAP

c7, of the signal sequence of Kaposi fibroblast growth factor oe JI

C35 Epitope Analog Exemplary Cell-Penetrating Exemplary Polypeptide Containing C35 Epitope Analog and CPP
Peptide Sequence (CPP) Analog of 1105- V113 of HSV-1 tegument protein VP22 IMNSRPPAVMTSRRSVKSGPREVPRDEYEDLYYTPSSGMASPDSPPDTSRRGALQTRSRQRGE
SEQ ID NO:2 having VRFVQYDESDYALYGGSSSEDDEHPEVPRTRRPVSGAVLSGPGPARAPPPPAGSGGAGRTPTT
tµ.) methionine substituted for APRAPRTQRVATKAPAAPAAE
ll'RGRKSAQPESAALPDAPASTAPTRSKTPAQGLARKLHFST oe threonine at the second amino APPNPDAPWTPRVAGFNKRVFCAAVGRLAAMHARMAAVQLWDMSRPRTDEDLNELLGITTI
acid residue and alp -nine RVTVCEGKNLLQRANELVNPDVVQDVDAATATRGRSAASRPTERPRAPARSASRPRRPVE, substituted for cysteine at the eighth amino acid residue MTSRRSVKSGPREVPRDEYEDLYYTPSSGMASPD SPPDTSRRGALQTRSRQRGEVRFVQYDES
(IMNSRPPCV) DYALYGGSSSEDDEHPEVPRTRRPVSGAVLSGPGPARAPPPPAGSGGAGRTPTTAPRAPRTQR
VATICAPAAPAAETTRGRKSAQPESAALPDAPASTAPTRSKTPAQGLARKLHFSTAPPNPDAPW
TPRVAGFNICRVFCAAVGRLAAMHARMAAVQLWDMSRPRTDEDLNELLGITTIRVTVCEGKN
LLQRANELVNPDVVQDVDAATATRGRSAASRPTERPRAPARSASRPRRPVEIMNSRPPAVMTS
RRSVKSGPREVPRDEYEDLYYTPSSGMASPDSPPDTSRRGALQTRSRQRGEVRFVQYDESDYA

LYGGSSSEDDEHPEVPRTRRPVSGAVLSGPGPARAPPPPAGSGGAGRTPTTAPRAPRTQRVATK
co c7, co ANELVNPDVVQDVDAATATRGRSAASRPTERPRAPARSASRPRRPVE

membrane-translocating IMNSRPPAVAAVLLPVLLAAP, sequence (MST) from h region AAVLLPVLLAAPIMNSRPPAVAAVLLPVLLAAP

of the signal sequence of Kaposi c7, fibroblast growth factor = oe JI

Analog of 1105 - V113 of HSV-1 tegument protein VP22 ITNSRPPSVMTSRRSVKSGPREVPRDEYEDLYYTPSSGMASPDSPPDTSRRGALQTRSRQRGEV
SEQ ID NO:2 having serine RFVQYDESDYALYGGS S
SEDDEHPEVPRTRRPVSGAVLSGPGPARAPPPPAGSGGAGRTPTTA
substituted for cysteine at the PRAPRTQRVATKAPAAPAAETTRGRKSAQPESAALPDAPASTAPTRSKTPAQGLARKLHFSTA
eighth amino acid residue PPNPDAPWTPRVAGFNICRVFCAAVGRLAAMHARMAAVQLWDMSRPRTDEDLNELLGITTIR
(ITNSRPPSV) VTVCEGKNLLQRANELVNPDVVQDVDAATATRGRSAASRPTERPRAPARSASRPRRPVE, tµ.) oe MTSRRSVKSGPREVPRDEYEDLYYTPSSGMASPDSPPDTSRRGALQTRSRQRGEVRFVQYDES
DYALYGGSSSEDDEHPEVPRTRRPVSGAVLSGP GPARAPPPPAGSGGAGRTPTTAPRAPRTQR
VATKAPAAPAAETTRGRKSAQPESAALPDAPASTAPTRSKTPAQGLARKLHFSTAPPNPDAPW
TPRVAGENKRVFCAAVGRLAAMHARMAAVQLWDMSRPRTDEDLNELLGITTIRVTVCEGKN
LLQRANELVNPDVVQDVDAATATRGRSAASRPTERPRAPARSASRPRRPVEITNSRPPSVMTSR
RSVKSGPREVPRDEYEDLYYTPSSGMASPDSPPDTSRRGALQTRSRQRGEVRFVQYDESDYAL
YGGSSSEDDEHPEVPRTRRPVSGAVLSGP GPARAPPPPAGSGGAGRTPTTAPRAPRTQRVATK
APAAPAAETTRGRKSAQPESAALPDAPASTAPTRSKTPAQGLARKLHFSTAPPNPDAPWTPRV

AGFNKRVFCAAVGRLAAMHARMAAVQLWDMSRPRTDEDLNELLGITTIRVTVCEGKNLLQR
ANELVNPDVVQDVDAATATRGRSAASRPTERPRAPARSASRPRRPVE
co co c7, co membrane-translocating ITNSRPPSVAAVLLPVLLAAP, sequence (MST) from h region AAVLLPVLLAAPITNSRPPSVAAVLLPVLLAAP

of the signal sequence of Kaposi fibroblast growth factor c7, JI
oe Analog of K104 - V113 of HSV-1 tegument protein VP22 K.ITNSRPPSVMTSRRSVKSGPREVPRDEYEDLYYTPSSGMASPDSPPDTSRRGALQTRSRQRGE
SEQ ID NO:2 having serine VRFVQYDESDYALYGGS S SEDD
EHPEVPRTRRPV S GAVLS GP GPARAPPPPAGS GGA GRTPTT
substituted for cysteine at the APRAPRTQRVATKAPAAPAAETTRGRKSAQPESAALPDAPASTAPTRSKTPAQGLARKLHFST
ninth amino acid residue APPNPDAPWTPRVAGFNKRVFCAAVGRLAAMHARMAAVQLWDMSRPRIDEDLNELLGITTI
(KITNSRPPSV) RVTVCEGKNLLQRANELVNPDVVQDVDAATATRGRSAASRPTERPRAPARSASRPRRPVE, tµ.) MTSRRSVKSGPREVPRDEYEDLYYTP SS GMA SPD SPPD TSRRGALQTRSRQRGEVRFVQYDES
DYALYGGS S SEDDEHPEVPRTRRPV S GAVLS GP GPARAPPPPA GS GGA GRTPTTAPRAPRTQR
VATKAPAAPAAETTRGRKSAQPESAALPDAPASTAPTRSKTPAQGLARKLHFSTAPPNPDAPW
TPRVAGFNKRVFCAAVGRLAAMHARMAAVQLWDMSRPRTDEDLNELLGITTIRVTVCEGKN
LLQRANEL VNPDVVQD VDAATATRGRSAA SRPTERPRAPARSA SRPRRPVEKITNSRPP SVMT
SRRSVKSGPREVPRDEYEDLYYTPSSGMASPDSPPDTSRRGALQTRSRQRGEVRFVQYDESDY
ALYGGS SSEDDEHPEVPRTRRPVS GAVLS GP GPARAPPPPAGS GGAGRTPTTAPRAPRTQRVAT
KAPAAPAAETTRGRKSAQPESAALPDAPASTAPTRSKTPAQGLARKLHFSTAPPNPDAPWTPR

VAGFNKRVFCAAVGRLAAMHARMAAVQLWDMSRPRTDEDLNELLGITTM.VTVCEGKNLLQ
co RANELVNPDVVQDVDAATATRGRSAASRPTERPRAPARSASRPRRPVE
co c7, co membrane-translocating KITNSRPP SVAAVLLPVLLAAP, Isis) sequence (MST) from h region AAVLLPVLLAAPKITNSRPPSVAAVLLPVLLAAP

of the signal sequence of Kaposi fibroblast growth factor JI
c7, oe Analog of G22 to C30 of SEQ HSV-1 tegument protein VP22 GVRIVVEYAMTSRRSVKSGPREVPRDEYEDLYYTPSSGMASPDSPPDTSRRGALQTRSRQRGE
ID NO:2 having alanine VRFVQYDESDYALYGGSSSEDDEHPEVPRTRRPVSGAVLSGPGPARAPPPPAGSGGAGRTPTT
substituted for cysteine at APRAPRTQRVATKAPAAPAAETTRGRKSAQPESAALPDAPASTAPTRSKTPAQGLARKLHFST
ninth amino acid residue APPNPDAPWTPRVAGFNKRVFCAAVGRLAAMHARMAAVQLWDMSRPRTDEDLNELLGITTI
(GVRIVVEYA) RVTVCEGKNLLQRANELVNPDVVQDVDAATATRGRSAASRPTERPRAPARSASRPRRPVE, tµ.) MTSRRSVKSGPREVPRDEYEDLYYTPSSGMASPDSPPDTSRRGALQTRSRQRGEVRFVQYDES
DYALYGGSSSEDDEHPEVPRTRRPVSGAVLSGP GPARAPPPPAGSGGAGRTPTTAPRAPRTQR
VATKAPAAPAAETTRGRKSAQPESAALPDAPASTAPTRSKTPAQGLARKLHFSTAPPNPDAPW
TPRVAGFNKRVFCAAVGRLAAMHARMAAVQLWDMSRPRTDEDLNELLGITTIRVTVCEGKN
LLQRANELVNPDVVQDVDAATATRGRSAASRPTERPRAPARSASRPRRPVEGVRIVVEYAMTS
RRSVKSGPREVPRDEYEDLYYTPSSGMASPDSPPDTSRRGALQTRSRQRGEVRFVQYDESDYA
LYGGSSSEDDEHPEVPRTRRPVSGAVLSGPGPARAPPPPAGSGGAGRTPTTAPRAPRTQRVATK
APAAPAAETTRGRKSAQPESAALPDAPAS TAPTRSKTPAQGLARKLHFSTAPPNPDAPWTPRV

AGFNKRVFCAAVGRLAAMHARMAAVQLWDMSRPRTDEDLNELLGITTIRVTVCEGKNLLQR
co ANELVNPDVVQDVDAATATRGRSAASRPTERPRAPARSASRPRRPVE
co c7, t!..) co membrane-translocating GVRTVVEYAAAVLLPVLLAAP, t=Q
sequence (MST) from h region AAVLLPVLLAAPGVRIVVEYAAAVLLPVLLAAP

of the signal sequence of Kaposi fibroblast growth factor JI
c7, oe Analog of 125 to C33 of SEQ HSV-1 tegument protein VP22 IVVEYAEPAMTSRRSVKSGPREVPRDEYEDLYYTPSSGMASPD SPPDTSRRGALQTRSRQRGE
ID NO:2 having alanine VRFVQYDESDYALYGGSSSEDDEHPEVPRTRRPVSGAVLSGPGPARAPPPPAGSGGAGRTPTT
substituted for cysteine at the APRAPRTQRVATICAPAAPAAETTRGRKSAQPESAALPDAPASTAPTRSKTPAQGLARKLHFST
sixth and ninth amino acid APPNPDAPWTPRVAGFNKRVFCAAVGRLAAMHARMAAVQLWDMSRPRTDEDLNELLGITTI
residues (IVVEYAEPA) RVTVCEGKNLLQRANELVNPDVVQDVDAATATRGRSAASRPTERPRAPARSASRPRRPVE, tµ.) oe MTSRRSVKSGPREVPRDEYEDLYYTPSSGMASPDSPPDTSRRGALQTRSRQRGEVRFVQYDES
DYALYGGSSSEDDEHPEVPRTRRPVSGAVLSGPGPARAPPPPAGSGGAGRTPTTAPRAPRTQR
VATKAPAAPAAETTRGRKSAQPESAALPDAPASTAPTRSKTPAQGLARKLHFSTAPPNPDAPW
TPRVAGFNKRVFCAAVGRLAAMHARMAAVQLWDMSRPRTDEDLNELLGITTIRVTVCEGKN
LLQRANELVNPDVVQDVDAATATRGRSAASRPTERPRAPARSASRPRRPVEIVVEYAEPAMTS
RRSVKSGPREVPRDEYEDLYYTPSSGMASPDSPPDTSRRGALQTRSRQRGEVRFVQYDESDYA
LYGGSSSEDDEHPEVPRTRRPVSGAVLSGPGPARAPPPPAGSGGAGRTPTTAPRAPRTQRVATK
APAAPAAETTRGRKSAQPESAALPDAPASTAPTRSKTPAQGLARKLHFSTAPPNPDAPWTPRV

AGFNKRVFCAAVGRLAAMHARMAAVQLWDMSRPRTDEDLNELLGITTIRVTVCEGKNLLQR
co ANELVNPDVVQDVDAATATRGRSAASRPTERPRAPARSASRPRRPVE
co c7, te) co membrane-translocating IVVEYAEPAAAVLLPVLLAAP, sequence (MST) from h region AAVLLPVLLAAPIVVEYAEPAAAVLLPVLLAAP

of the signal sequence of Kaposi fibroblast growth factor JI
c7, oe Analog of K104 -C112 of HSV-1 tegument protein VP22 KITNSRPPAMTSRRSVKSGPREVPRDEYEDLYYTPSSGMASPDSPPDTSRRGALQTRSRQRGEV
SEQ ID NO:2 having alanine RFVQYDESDYALYGGS S
SEDDEHPEVPRTRRPVSGAVLSGP GPARAPPPPA GS GGA GRTPTTA
substituted for cysteine at the PRAPRTQRVATKAPAAPAAETTRGRKSAQPESAALPDAPASTAPTRSKTPAQGLARKLHFSTA
ninth amino acid residue PPNPDAPWTPRVAGFNKRVFCAAVGRLAAMHARMAAVQLWDMSRPRTDEDLNELLGITTIR
(K1TNSRPPA) VTVCEGKNLLQRANELVNPDVVQDVDAATA
l'KGRSAASRPTERPRAPARSASRPRRPVE, tµ.) oe MTSRRSVKSGPREVPRDEYEDLYYTPSSGMASPDSPPDTSRRGALQTRSRQRGEVRFVQYDES
DYALYGGSSSEDDEHPEVPRTRRPVSGAVLSGPGPARAPPPPAGSGGAGRTPTTAPRAPRTQR
VATKAPAAPAAETTRGRKSAQPESAALPDAPASTAPTRSKTPAQGLARKLHFSTAPPNPDAPW
TPRVAGFNKRVFCAAVGRLAAMHARMAAVQLWDMSRPRTDEDLNELLGITTIRVTVCEGKN
LLQRANELVNPDVVQDVDAATATRGRSAASRPTERPRAPARSASRPRRPVEKITNSRPPAMTS
RRSVKSGPREVPRDEYEDLYYTPSSGMASPDSPPDTSRRGALQTRSRQRGEVRFVQYDESDYA
LYGGSSSEDDEHPEVPRTRRPVSGAVLSGPGPARAPPPPAGSGGAGRTPTTAPRAPRTQRVATK
APAAPAAETTRGRICSAQPESAALPDAPASTAPTRSKTPAQGLARKLHFSTAPPNPDAPWTPRV

AGFNICRVFCAAVGRLAAMHARMAAVQLWDMSRPRTDEDLNELLGITTIRVTVCEGKNLLQR
co ANELVNPDVVQDVDAATATRGRSAASRPTERPRAPARSASRPRRPVE
co c7, ts!..) co membrane-translocating KITNSRPPAAAVLLPVLLAAP, sequence (MST) from h region AAVLLPVLLAAP KITNSRPPAAAVLLPVLLAAP

of the signal sequence of Kaposi fibroblast growth factor JI
c7, oe Analog of K104 -V113 of HSV-1 tegument protein VP22 KITNSRPPAVMTSRRSVKSGPREVPRDEYEDLYYTPSSGMASPDSPPDTSRRGALQTRSRQRGE
SEQ ID NO:2 having alanine VRFVQYDESDYALYGGSSSEDDEHPEVPRTRRPVSGAVLS GP GPARAPPPPAGS GGAGRTPTT
substituted for cysteine at the APRAPRTQRVATKAPAAPAAETTRGRKSAQPESAALPDAPASTAPTRSKTPAQGLARKLHF ST
ninth amino acid residue APPNPDAPWTPRVAGFNKRVF
CAAVGRLAAMHARMAAVQLWDMSRPRTDEDLNELLGITTI
(KITNSRPPAV) RVTVCEGKNLLQRANELVNPDVVQDVDAATATRGRSAASRPTERPRAPARSASRPRRPVE, tµ.) oe MTSRRSVKSGPREVPRDEYEDLYYTPSSGMASPDSPPDTSRRGALQTRSRQRGEVRFVQYDES
DYALYGGSSSEDDEHPE VPRTRRPVS GAVLSGP GPARAPPPPAGSGGAGRTPTTAPRAPRTQR
VATKAPAAPAAETTRGRKSAQPESAALPDAPASTAPTRSKTPAQGLARKLHFSTAPPNPDAPW
TPRVAGFNKRVFCAAVGRLAAMHARMAAVQLWDMSRPR'TDEDLNELLGITTIRVTVCEGKN
LLQRANELVNPDVVQDVDAATATRGRSAASRPTERPRAPARSASRPRRPVEKITNSRPPAVMT
SRRSVKSGPREVPRDEYEDLYYTP SSGMASPDSPPDTSRRGALQTRSRQRGEVRFVQYDESDY
ALYGGSSSEDDEHPEVPRTRRPVSGAVLSGPGPARAPPPPAGSGGAGRTPTTAPRAPRTQRVAT
KAPAAPAAETTRGRKSAQPESAALPDAPASTAPTRSKTPAQGLARKLHFSTAPPNPDAPWTPR

VAGFNKRVFCAAVGRLAAMHARMAAVQLWDMSRPRTDEDLNELLGITTIRVTVCEGKNLLQ
RANELVNPDVVQDVDAATATRGRSAASRPTERPRAPARSASRPRRPVE
t:) membrane-translocating KITNSRPPAVAAVLLPVLLAAP, N.) sequence (MST) from h region AAVLLPVLLAAP KITNSRPPAVAAVLLPVLLAAP

of the signal sequence of Kaposi fibroblast growth factor JI
oe [0266] The peptides in accordance with the invention can be a variety of lengths, and either in their neutral (uncharged) forms or in forms which are salts. The peptides in accordance with the invention can contain modifications such as glycosylation, side chain oxidation, or phosphorylation, generally subject to the condition that modifications do not destroy the biological activity of the peptides.
[0267] The peptides of the invention can be prepared in a wide variety of ways.
For the preferred relatively short size, the peptides can be synthesized in solution or on a solid support in accordance with conventional techniques. Various automatic synthesizers are commercially available and can be used in accordance with known protocols. (See, for example, Stewart & Young, SOLID PHASE
PEPTIDE SYNTHESIS, 2D. ED., Pierce Chemical Co., 1984). Further, individual C35 peptide epitopes and C35 peptide epitope analogs can be joined using chemical ligation to produce larger homopolymer or heteropolymer polypeptides that are still within the bounds of the invention.
[0268] Alternatively, recombinant DNA technology can be employed wherein a nucleotide sequence which encodes an immunogenic peptide of interest is inserted into an expression vector, transformed or transfected into an appropriate host cell and cultivated under conditions suitable for expression. These procedures are generally known in the art, as described generally in Sambrook et al., MOLECULAR CLONING, A LABORATORY MANUAL, Cold Spring Harbor Press, Cold Spring Harbor, New York (1989). Thus, recombinant polypepfides, which comprise one or more peptide epitope sequences of the invention, can be used to present the appropriate T cell epitope.
[0269] The nucleotide coding sequence for C35 peptide epitopes or C35 peptide epitope analogs of the preferred lengths contemplated herein can be synthesized by chemical techniques, for example, the phosphotriester method of Matteucci, etal., J. Am. Chem. Soc. 103:3185 (1981). Peptide analogs can be made simply by substituting the appropriate and desired nucleic acid base(s) for those that encode the native peptide sequence; exemplary nucleic acid substitutions are those that encode an amino acid defmed by the motifs/supermotifs herein. The coding sequence can then be provided with appropriate linkers and ligated into expression vectors commonly available in the art, and the vectors used to transform suitable hosts to produce the desired fusion protein. A number of such vectors and suitable host systems are now available. For expression of the fusion proteins, the coding sequence will be provided with operably linked start and stop codons, promoter and terminator regions and usually a replication system to provide an expression vector for expression in the desired cellular host. For example, promoter sequences compatible with bacterial hosts are provided in plasmids containing convenient restriction sites for insertion of the desired coding sequence. The resulting expression vectors are transformed into suitable bacterial hosts. Of course, yeast, insect or mammalian cell hosts may also be used, employing suitable vectors and control sequences.
[02701 It is generally preferable that the peptide epitope be as small as possible while still maintaining substantially all of the immunologic activity of the native protein. When possible, it may be desirable to optimize HLA class I binding peptide epitopes of the invention to a length of about 8 to about 13 amino acid residues, preferably 9 to 10. It is to be appreciated that a longer polyp eptide, e.g., a C35 polypeptide fragment or a synthetic polypeptide, can comprise one or more C35 peptide epitopes or C35 peptide epitope analogs in this size range (see the Definition Section for the term "epitope" for further discussion ofpeptide length).
HLA class II binding epitopes are preferably optimized to a length of about 6 to about 30 amino acids in length, preferably to between about 13 and about 20 residues. Preferably, the epitopes are commensurate in size with endogenously processed pathogen-derived peptides or tumor cell peptides that are bound to the relevant HLA molecules. The identification and preparation ofpeptides ofvarious lengths can be carried out using the techniques described herein.
[02711 An alternative preferred embodiment of the invention comprises administration of peptides of the invention linked as a polyepitopic polypeptide, e.g., homopolymers or heteropolymers, or as a minigene that encodes a polyepitopic polypeptide.

[02721 Another preferred embodiment is obtained by identifying native C35 polypeptide regions that contain a high concentration of class I and/or class peptide epitopes. Such a sequence is generally selected on the basis that it contains the greatest number of C35 epitopes per amino acid length. It is to be appreciated that epitopes can be present in a frame-shifted manner, e.g. a 10 amino acid long peptide could contain two 9 amino acid long epitopes and one amino acid long epitope; upon intracellular processing, each epitope can be exposed and bound by an HLA molecule upon administration of such a peptide.
Thus a larger, preferably multi-epitopic, polypeptide can be generated synthetically, recombinantly, or via cleavage from the native source.
Assays to Detect T-Cell Responses [0273] Once HLA. binding peptides are identified, they can be tested for the ability to elicit a T-cell response. The preparation and evaluation ofmotif-bearing peptides are described, e.g., in PCT publications WO 94/20127 and WO
94/03205.
Briefly, peptides comprising epitopes from a particular antigen are synthesized and tested for their ability to bind to relevant HLA proteins. These assays may involve evaluation of peptide binding to purified HLA class I molecules in relation to the binding of a radioiodinated reference peptide. Alternatively, cells expressing empty class I molecules e. cell surface HLA molecules that lack any bound peptide) may be evaluated for peptide binding by immundluorescent staining and flow microfluorimetry. Other assays that may be used to evaluate peptide binding include peptide-dependent class I assembly assays and/or the inhibition of CTL recognition by peptide competition. Those peptides that bind to an HLA class I molecule, typically with an affinity of 500 nM or less, are further evaluated for their ability to serve as targets for CTLs derived from infected or immilni zed individuals, as well as for their capacity to induce primary in vitro or in vivo CTL responses that can give rise to CTL populations capable of reacting with selected target cells associated with pathology.
[0274] Analogous assays are used for evaluation of HLA class II binding peptides. HLA class II motif-bearing peptides that are shown to bind, typically at an affinity of 1000 nM or less, are further evaluated for the ability to stimulate HTL responses.
[0275] Conventional assays utilized to detect T cell responses include proliferation assays, lymphokine secretion assays, direct cytotoxicity assays, and limiting dilution assays. For example, antigen-presenting cells that have been incubated with a peptide can be assayed for the ability to induce CTL
responses in responder cell populations. Antigen-presenting cells can be nounal cells such as peripheral blood mononuclear cells or dendritic cells. Alternatively, mutant, non-human mammalian cell lines that have been transfected with a human class I

MHC gene, and that are deficient in their ability to load class I molecules with internally processed peptides, are used to evaluate the capacity of the peptide to induce in vitro primary CTL responses. Peripheral blood mononuclear cells (PBMCs) can be used as the source of CTL precursors. Antigen presenting cells are incubated with peptide, after which the peptide-loaded antigen-presenting cells are then incubated with the responder cell population under optimized culture conditions. Positive CTL activation can be determined by assaying the culture for the presence of CTLs that lyse radio-labeled target cells, either specific peptide-pulsed targets or target cells that express endogenously processed antigen from which the specific peptide was derived. Alternatively, the presence of epitope-specific CTLs can be determined by IENy in situ ELISA.
[0276] Additionally, a method has been devised which allows direct quantification of antigen-specific T cells by staining with fluorescein-labelled HLA tetrameric complexes (Altman, J. D. et al., Proc. Natl. Acad. Sci. USA
90:10330, 1993; Altman, J. D. et al., Science 274:94, 1996). Other options include staining for intracellular lymphokines, and interferon release assays or ELISPOT assays. Tetramer staining, intracellular lympholdne staining and =

ELISPOT assays all appear to be at least 10-fold more sensitive than more conventional assays (Lalvani, A. et al., J. Exp. Med. 186:859, 1997; Dunbar, P.
R. et al., Curr. Biol. 8:413, 1998; Murali-Krishna, K. et al., Immunity 8:177, 1998).
[0277] Helper T lymphocyte (HTL) activation may also be assessed using techniques known to those in the art, such as T cell proliferation or lymphokine secretion (see, e.g. Alexander et al., Immunity 1:751-761, 1994).
[0278] Alternatively, immunization of HLA transgenic mice can be used to determine immunogenicity ofpeptide epitopes. Several transgenic mouse strains, e.g., mice with human A2.1, All (which can additionally be used to analyze HLA-A3 epitopes), and B7 alleles have been characterized. Other transgenic mice strains (e.g., transgenic mice for HLA-Al and A24) are being developed.
Moreover, HLA-DR1 and HLA-DR3 mouse models have been developed. In accordance with principles in the art, additional transgenic mouse models with other HLA alleles are generated as necessary.
[0279] Such mice can be immunized with peptides emulsified in Incomplete Freund's Adjuvant; thereafter any resulting T cells can be tested for their capacity to recognize target cells that have been peptide-pulsed or transfected with genes encoding the peptide of interest. CTL responses can be analyzed using cytotoxicity assays described above. Similarly, HTL responses can be analyzed using, e.g., T cell proliferation or lympholdne secretion assays.
Vaccine Compositions [0280] Vaccines that contain an immunologically effective amount of one or more C35 peptide epitopes and/or C35 peptide epitope analogs of the invention are a further embodiment of the invention. The peptides can be delivered by various means or formulations, all collectively referred to as "vaccine"
compositions. Such vaccine compositions, and/or modes of administration, can include, for example, naked cDNA in cationic lipid formulations; lipopeptides (e.g.,Vitiello, A. et al., J. Clin. Invest. 95:341, 1995), naked cDNA or peptides, encapsulated e.g., in poly(DL-lactide-co-glycolide) ("PLG") microspheres (see, e.g., Eldridge, et al., Molec. Immunol. 28:287-294, 1991: Alonso et al., Vaccine 12:299-306, 1994; Jones et al.,Vaccine 13:675-681, 1995); peptide compositions contained in immune stimulating complexes (ISCOMS) (see, e.g., Takahashi et al. ,Nature 344:873-875, 1990; Hu et al., Clin Exp Immunol. 113:235-243,1998);

multiple antigen peptide systems (MAPs) (see e.g., Tam, J. P., Proc. Natl.
Acad.
Sci. U.S.A. 85:5409-5413, 1988; Tam, J.P., J. Immunol. Methods 196:17-32, 1996); viral, bacterial, or, fungal delivery vectors (Perkus, M. E. et al., In:
Concepts in vaccine development, Kaufmann, S. H. E., ed., p. 379, 1996;
Chakrabarti, S. et al., Nature 320:535, 1986; Hu, S. L. et al., Nature 320:537, 1986; Kieny, M.-P. et al., AIDS Bio/Technology 4:790, 1986; Top, F. H. etal., J. Infect. Dis. 124:148, 1971; Chanda, P. K. et al., Virology 175:535, 1990);
particles of viral or synthetic origin (e.g., Kofler, N. etal., J. Immunol.
Methods.
192:25, 1996; Eldridge, J. H. et al., Sem. Hematol. 30:16, 1993; Falo, L. D., Jr.
et al., Nature Med. 7:649, 1995); adjuvants (Warren, H. S., Vogel, F. R., and Chedid, L. A. Annu. Rev. Immunol. 4:369, 1986; Gupta, R. K. et al., Vaccine 11:293, 1993); liposomes (Reddy, R. etal., J. Immunol. 148:1585, 1992; Rock, K. L., Immunol. Today 17:131, 1996); or, particle-absorbed cDNA (Ulmer, J. B.
etal., Science 259:1745, 1993; Robinson, H. L., Hunt, L. A., and Webster, R.
G., Vaccine 11:957, 1993; Shiver, J. W. etal., In: Concepts in vaccine development, Kaufmann, S. H. E., ed., p. 423, 1996; Cease, K. B., and Berzofsky, J. A., Annu.
Rev. Immunol. 12:923, 1994 and Eldridge, J. H. et al., Sem. Hematol. 30:16, = 1993), etc. Toxin-targeted delivery technologies, also known as receptor mediated targeting, such as those of Avant Immunotherapeutics, Inc. (Needham, Massachusetts) or attached to a stress protein, e.g., HSP 96 (Stressgen Biotechnologies Corp., Victoria, BC, Canada) can also be used.
[0281] Vaccines of the invention comprise nucleic acid mediated modalities.
DNA or RNA encoding one or more of the polypeptides of the invention can be administered to a patient. This approach is described, for instance, in Wolff et.

al., Science 247:1465 (1990) as well as U.S. Patent Nos. 5,580,859; 5,589,466;

5,804,566; 5,739,118; 5,736,524; 5,679,647; and, WO 98/04720. Examples of DNA-based delivery technologies include "naked DNA", facilitated (bupivicaine, polymers, peptide-mediated) delivery, cationic lipid complexes, and particle-mediated ("gene gun") or pressure-mediated delivery (see, e.g., U.S. Patent No.
5,922,687). Accordingly, peptide vaccines of the invention can be expressed by viral or bacterial vectors. Examples of expression vectors include attenuated viral hosts, such as vaccinia or fowlpox. For example, vaccinia virus is used as a vector to express nucleotide sequences that encode the peptides of the invention.
Upon introduction into an acutely or chronically infected host or into a non-infected host, the recombinant vaccinia virus expresses the immunogenic peptide, and thereby elicits an immune response. Vaccinia vectors and methods useful in immunization protocols are described in, e.g., U.S. Patent No. 4,722,848.
Another vector is BCG (Bacille Calmette Guerin). BCG vectors are described in Stover et al. ,Nature 351:456-460(1991). A wide variety of other vectors useful for therapeutic administration or immunization of the peptides of the invention, e.g. adeno and adeno-associated virus vectors, alpha virus vectors, retroviral vectors, Salmonella typhi vectors, detoxified anthrax toxin vectors, and the like, are apparent to those skilled in the art from the description herein.
[0282] Furthermore, vaccines in accordance with the invention can comprise one or more C35 peptide epitopes of the invention. Accordingly, a C35 peptide epitope or C35 peptide epitope analog can be present in a vaccine individually or;
alternatively, the peptide epitope or analog can exist as multiple copies of the same peptide epitope or analog (a homopolymer), or as multiple different peptide epitopes or analogs (a heteropolymet). Polymers have the advantage of increased probability for immunological reaction and, where different peptide epitopes or analogs are used to make up the polymer, the ability to induce antibodies and/or T cells that react with different antigenic determinants of the antigen targeted for an immune response. The composition may be a naturally occurring region of an antigen or can be prepared, e.g., recombinantly or by chemical synthesis.

[0283] Carriers that can be used with vaccines of the invention are well known in the art, and include, e.g., thyroglobulin, albumins such as human serum albumin, tetanus toxoid, polyamino acids such as poly L-lysine, poly L-glutamic acid, influenza virus proteins, hepatitis B virus core protein, and the like.
The vaccines can contain a physiologically tolerable diluent such as water, or a saline solution, preferably phosphate buffered saline. Generally, the vaccines also include an adjuvant. Adjuvants such as incomplete Freund's adjuvant, aluminum phosphate, aluminum hydroxide, or alum are examples of materials well known in the art. Additionally, as disclosed herein, CTL responses can be primed by conjugating peptides of the invention to lipids, such as tripalmitoyl-S-glyceryl-cysteinyl-seryl-serine (P3CSS).
[0284] Upon immunization with a peptide composition in accordance with the invention, via injection (e.g., subcutaneous, intradermal, intramuscular, aerosol, oral, transdermal, transmucosal, intrapleural, intrathecal), or other suitable routes, the immune system of the host responds to the vaccine by producing antibodies, CTLs and/or HTLs specific for the desired antigen. Consequently, the host becomes at least partially immune to subsequent exposure to the TAA, or at least partially resistant to further development of tumor associated antigen-bearing cells and thereby derives a prophylactic or therapeutic benefit.
[0285] In certain embodiments, components that induce T cell responses are combined with components that induce antibody responses to the target antigen of interest. A preferred embodiment of such a composition comprises class I
and class II epitopes in accordance with the invention. Alternatively, a composition comprises a class I and/or class II epitope in accordance with the invention, along with a PADRETM molecule (Epimmune, San Diego, CA).
[0286] Vaccine compositions of the invention can comprise antigen presenting cells, such as dendritic cells, as a vehicle to present peptides of the invention. For example, dendritic cells are transfected, e.g., with a minigene construct in accordance with the invention, in order to elicit immune responses. Minigenes are discussed in greater detail in a following section. Vaccine compositions can be created in vitro, following dendritic cell mobilization and harvesting, whereby loading of dendritic cells occurs in vitro.
[0287] The vaccine compositions of the invention may also be used in combination with antiviral drugs such as interferon-a, or immune adjuvants such as IL-12, GM-CSF, etc.
[0288]
Preferably, the following principles are utilized when selecting epitope(s) for inclusion in a vaccine, either peptide-based or nucleic acid-based formulations. Each of the following principles can be balanced in order to make the selection. When multiple epitopes are to be used in a vaccine, the epitopes may be, but need not be, contiguous in sequence in the native antigen from which the epitopes are derived.
[0289] 1) Epitopes are selected which, upon administration, mimic immune responses that have been observed to be correlated with prevention or clearance of TAA-expressing tumors. For HLA Class I, this generally includes 3-4 epitopes derived from at least one TAA.
[0290] 2) Epitopes are selected that have the requisite binding affinity established to be correlated with immunogenicity: for HLA Class I an IC50 of nM or less, or for Class II an IC50 of 1000 n1\4 or less. For HLA Class lit is presently preferred to select a peptide having an IC50 of 200 nM or less, as this is believed to better correlate not only to induction of an immune response, but to in vitro tumor cell killing as well.
[0291] 3) Supermotifbearing-peptides, or a sufficient array of allele-specific motif-bearing peptides, are selected to give broad population coverage. In general, it is preferable to have at least 80% population coverage. A Monte Carlo analysis, a statistical evaluation known in the art, can be employed to assess the breadth of population coverage.
[0292] 4) When selecting epitopes from cancer-related antigens, it can be preferable to include analog peptides in the selection, because the patient may have developed tolerance to the native epitope. When selecting epitopes for infectious disease-related antigens it is presently preferable to select either native or analog epitopes.
[0293] 5) Of particular relevance are "nested epitopes." Nested epitopes (or epitope analogs) occur where at least two epitopes or analogs (or an epitope and an analog) overlap in a given polypeptide sequence. A polypeptide comprising "transcendent nested epitopes" is a polypeptide that has both HLA class I and HLA class II epitopes and/or analogs in it. When providing nested epitopes, it is preferable to provide a sequence that has the greatest number of epitopes or analogs per provided sequence. Preferably, one avoids providing a polyp eptide that is any longer than the combined length of the peptide epitopes or analogs.
When providing a polypeptide comprising nested epitopes, it is important to evaluate the polypeptide in order to insure that it does not have pathological or other deleterious biological properties; this is particularly relevant for vaccines directed to infectious organisms. Thus, in a preferred embodiment, the vaccine compositions of the invention comprise one or more multi-epitope polypeptides selected from the group consisting of: 1105 to V113 of SEQ ID NO:2 and FIG.
1B, T101 to V113 of SEQ NO:2 and FIG. 1B, E100 to V113 of SEQ ID NO:2 and FIG. 1B, G99 to V113 of SEQ ID NO:2 and FIG. 1B, 193 to V113 of SEQ ID
NO:2 and FIG. 1B, D88 to V113 of SEQ ID NO:2 and FIG. 1B, P84 to V113 of SEQ ID NO:2 and FIG. 1B, K77 to V113 of SEQ ID NO:2 and FIG. 1B, Q72 to V113 of SEQ ID NO:2 and FIG. 1B, F65 to V113 of SEQ ID NO:2 and FIG. 1B, and L59 to V113 of SEQ ID NO:2 and FIG. 1B.
[0294] 6) If a polypeptide comprising more than one C35 peptide epitope or C35 peptide epitope analog is created, or when creating a minigene, an objective is to generate the smallest polypeptide that encompasses the epitopes/analogs of interest. This principle is similar, if not the same as that employed when selecting a polypeptide comprising nested epitopes. However, with an artificial polyepitopic polypeptide, the size minimization objective is balanced against the need to integrate any spacer sequences between epitopes in the polyepitopic polypeptide. Spacer or linker amino acid residues can be introduced to avoid junctional epitopes (an epitope recognized by the immune system, not present in the target antigen, and only created by the man-made juxtaposition of epitopes), or to facilitate cleavage between epitopes and thereby enhance epitope presentation. Junctional epitopes are generally to be avoided because the recipient may generate an immune response to that non-native epitope. Of particular concern is a junctional epitope that is a "dominant epitope." A
dominant epitope may lead to such a zealous response that immune responses to other epitopes are diminished or suppressed.
Minigene Vaccines [0295] A number of different approaches are available which allow simultaneous delivery of multiple epitopes. Nucleic acids encoding multiple C35 peptide epitopes or analogs are a useful embodiment of the invention; discrete epitopes/analogs or polyepitopic polypeptides can be encoded. The epitopes or analogs to be included in a minigene are preferably selected according to the guidelines set forth in the previous section. Examples of amino acid sequences that can be included in a minigene include: HLA class I epitopes or analogs, HLA
class If epitopes or analogs, a ubiquitination signal sequence, and/or a targeting sequence such as an endoplasmic reticulum (ER) signal sequence to facilitate movement of the resulting peptide into the endoplasmic reticulum.
[0296] The use of multi-epitope minigenes is also described in, e.g., Ishioka et aL, J. ImmunoL 162:3915-3925, 1999; An, L. and Whitton, J. L., J. ViroL
71:2292, 1997; Thomson, S. A. etal., J. ImmunoL 157:822, 1996; Whitton, J. L.
et al., J. ViroL 67:348, 1993; Hanke, R. etal., Vaccine 16:426, 1998. A
similar approach can be used to develop rninigenes encoding TAA epitopes.
[0297] For example, to create a DNA sequence encoding the selected epitopes (minigene) for expression in human cells, the amino acid sequences of the epitopes may be reverse translated. A human codon usage table can be used to guide the codon choice for each amino acid. These epitope-encoding DNA

sequences may be directly adjoined, so that when translated, a continuous polypeptide sequence is created. However, to optimize expression and/or immunogenicity, additional elements can be incorporated into the minigene design such as one or more spacer or linker amino acid residues between epitopes. HLA presentation of CTL and HTL epitopes may be improved by including synthetic (e.g. poly-alanine) or naturally-occurring flanking sequences adjacent to the CTL or HTL epitopes; these larger polypeptides comprising the epitope(s)/analog(s) are within the scope of the invention.
[0298] The minigene sequence may be converted to DNA by assembling oligonucleotides that encode the plus and minus strands of the minigene.
Overlapping oligonucleotides (30-100 bases long) may be synthesized, phosphorylated, purified and annealed under appropriate conditions using well known techniques. The ends of the oligonucleotides can be joined, for example, using T4 DNA ligase. This synthetic minigene, encoding the epitopepolypeptide, can then be cloned into a desired expression vector.
[0299] Standard regulatory sequences well known to those of skill in the art are preferably included in the vector to ensure expression in the target cells.
Several vector elements are desirable: a promoter with a downstream cloning site for minigene insertion; a polyadenylation signal for efficient transcription termination; an E. coli origin of replication; and an E. coli selectable marker (e.g.
ampicillin or kanamycin resistance). Numerous promoters can be used for this purpose, e.g., the human cytomegalovirus (hCMV) promoter. See, e.g., U.S.
Patent Nos. 5,580,859 and 5,589,466 for other suitable promoter sequences.
[0300] Optimized peptide expression and imrnunogenicity can be achieved by certain modifications to a minigene construct. For example, in some cases introns facilitate efficient gene expression, thus one or more synthetic or naturally-occurring introns can be incorporated into the transcribed region of the minigene. The inclusion of mRNA stabilization sequences and sequences for replication in mammalian cells may also be considered for increasing minigene expression.

[03011 Once an expression vector is selected, the minigene is cloned into the polylinker region downstream of the promoter. This plasmid is transformed into an appropriate bacterial strain, and DNA is prepared using standard techniques.
The orientation and DNA sequence of the minigene, as well as all other elements included in the vector, are confirmed using restriction mapping and DNA
sequence analysis. Bacterial cells harboring the correct plasmid can be stored as cell banks.
[0302] In addition, immunostimulatory sequences (IS Ss or CpGs) appear to play a role in the immunogenicity of DNA vaccines. These sequences may be included in the vector, outside the minigene coding sequence to enhance immunogenicity.
[03031 In some embodiments, a bi-cistronic expression vector which allows production of both the minigene-encoded epitopes and a second protein (e.g., one that modulates immunogenicity) can be used. Examples of proteins or polypeptides that, if co-expressed with epitopes, can enhance an immune response include cytokines (e.g., IL-2, IL-12, GM-CSF), cytokine-inducing molecules (e.g., LelF), costimulatory molecules, or pan-DR binding proteins (ADRETM, Epimmune, San Diego, CA). Helper T cell (HTL) epitopes such as PAIDPETM molecules can be joined to intracellular targeting signals and expressed separately from expressed CTL epitopes. This can be done in order to direct HTL epitopes to a cell compartment different than that of the CTL
epitopes, one that provides for more efficient entry of HTL epitopes into the HLA
class 11 pathway, thereby improving HTL induction. In contrast to HTL or CTL
induction, specifically decreasing the immune response by co-expression of immtmosuppressive molecules (e.g. TGF-13) maybe beneficial in certain diseases.
[0304] Therapeutic quantities of plasmid DNA can be produced for example, by fermentation in E. coli, followed by purification. Aliquots from the working cell bank are used to inoculate growth medium, and are grown to saturation in shaker flasks or a bioreactor according to well known techniques. Plasmid DNA is purified using standard bioseparation technologies such as solid phase anion-exchange resins available, e.g., from QIAGEN, Inc. (Valencia, California). If required, supercoiled DNA can be isolated from the open circular and linear forms using gel electrophoresis or other methods.
[0305] Purified plasmid DNA can be prepared for injection using a variety of formulations. The simplest of these is reconstitution of lyophilized DNA in sterile phosphate-buffer saline (PBS). This approach, known as "naked DNA,"
is currently being used for intramuscular (IM) administration in clinical trials. To maximize the immunotherapeutic effects of minigene vaccines, alternative methods of formulating purified plasmid DNA may be used. A variety of such methods have been described, and new techniques may become available.
Cationic lipids, glycolipids, and fusogenic liposomes can also be used in the formulation (see, e.g., WO 93/24640; Mannino & Gould-Fogerite,BioTechniques 6(7): 682 (1988); U.S. Patent No. 5,279,833; WO 91/06309; and Feigner, et al., Proc. NatlAcad. Sci. USA 84:7413(1987). In addition, peptides and compounds referred to collectively as protective, interactive, non-condensing compounds (PINC) can also be complexed to purified plasmid DNA to influence variables such as stability, intramuscular dispersion, or trafficking to specific organs or cell types.
[0306] Target cell sensitization can be used as a functional assay of the expression and HLA class I presentation of minigene-encoded epitopes. For example, the plasmid DNA is introduced into a mammalian cell line that is a suitable target for standard CTL chromium release assays. The transfection method used will be dependent on the final formulation, electroporation can be used for "naked" DNA, whereas cationic lipids allow direct in vitro transfection.
A plasmid expressing green fluorescent protein (GFP) can be co-transfected to allow enrichment of transfected cells using fluorescence activated cell sorting (FACS). The transfected cells are then chromium-51 (51Cr) labeled and used as targets for epitope-specific CTLs. Cytolysis of the target cells, detected by 51Cr release, indicates both the production and HLA presentation of, minigene-encoded CTL epitopes. Expression of HTL epitopes may be evaluated in an analogous manner using assays to assess HTL activity.
[0307] In vivo immunogenicity is a second approach for functional testing of minigene DNA formulations. Transgenic mice expressing appropriate human HLA proteins are immunized with the DNA product. The dose and route of administration are formulation dependent (e.g., IM for DNA in PBS, intraperitoneal (IP) for lipid-complexed DNA). Eleven to twenty-one days after immunization, splenocytes are harvested and restimulated for one week in the presence of peptides encoding each epitope being tested. Thereafter, for CTLs, standard assays are conducted to determine if there is cytolysis ofpeptide-loaded, 'Cr-labeled target cells. Once again, lysis of target cells that were exposed to epitopes corresponding to those in the minigene, demonstrates DNA vaccine function and induction of CTLs. Immunogenicity of HTL epitopes is evaluated in transgenic mice in an analogous manner.
[0308] Alternatively, the nucleic acids can be administered using ballistic delivery as described, for instance, in U.S. Patent No. 5,204,253. Using this technique, particles comprised solely of DNA are administered. In a further alternative embodiment for ballistic delivery, DNA can be adhered to particles, such as gold particles.
Combinations of CTL Peptides with Helper Peptides [0309] Vaccine compositions comprising CTL peptides of the present invention can be modified to provide desired attributes, such as improved serum half-life, broadened population coverage or enhanced immunogenicity.
[0310] For instance, the ability of a peptide to induce CTL activity can be enhanced by linking the CTL peptide to a sequence which contains at least one HTL epitope.
[0311] Although a CTL peptide can be directly linked to a T helper peptide, particularlypreferred CTL epitope/HTL epitope conjugates are linked by a spacer molecule. The spacer is typically comprised of relatively small, neutral molecules, e.g., amino acids or amino acid mimetics, which are substantially uncharged under physiological conditions. The spacers are typically selected from, e.g., Ala, Gly, or other neutral spacers of nonpolar amino acids or neutral polar amino acids. It will be understood that the optional spacer need not be comprised of the same residues and thus may be a hetero- or homo-oligomer.
When present, the spacer will usually be at least one or two residues, commonly three to 13, more frequently three to six residues. The CTL peptide epitope may be linked to the T helper peptide epitope, directly or via a spacer, at either it's amino or carboxyl terminus. The amino terminus of either the CTL peptide or the HTL peptide can be acylated.
[03121 In certain embodiments, the T helper peptide is one that is recognized by T helper cells present in the majority of the population. This can be accomplished by selecting amino acid sequences that bind to many, most, or all of the HLA class II molecules. These are known as "loosely HLA-restricted" or "promiscuous" T helper sequences. Examples of amino acid sequences that are promiscuous include sequences from antigens such as tetanus toxoid at positions 830-843 (QYIKANSKFIGITE), Plasmodium fakzparurn CS protein at positions 378-398 (DIEKKIAKMEKASSVFNVVNS), and Streptococcus 181(.13 protein at positions 116 (GAVDSILGGVATYGAA). Other examples include peptides bearing a DR 1-4-7 supermotif, or either of the DR3 motifs.
[03131 Alternatively, it is possible to prepare synthetic peptides capable of stimulating T helper lymphocytes, in a loosely HLA-restricted fashion, using amino acid sequences that may not be found in nature. Synthetic compounds fall within the family of molecules called Pan-DR-binding epitopes (e.g., PADRE, Epimmune Inc., San Diego, CA). PADRETM peptides are designed to bind multiple HLA-DR (human HLA class II) molecules. For instance, a pan-DR-binding epitope peptide having the formula: alOCVAAZTLKAAa, where "X" is either cyclohexylalanine, phenylalanine, or tyrosine; "Z" is either tryptophan, tyrosine, histidine or asparagine; and "a" is either D-alanine or L-alanine, has been found to bind to numerous allele-specific HLA-DR molecules. Accordingly, these molecules stimulate a T helper lymphocyte response from most individuals, regardless of their HLA type. Certain pan-DR binding epitopes comprise all "L"

natural amino acids; these molecules can be provided as peptides or in the form of nucleic acids that encode the peptide.
[0314] HTL peptide epitopes can be modified to alter their biological properties.
HTL peptide epitopes can be modified in the same manner as CTL peptides. For instance, they may be modified to include D-amino acids or be conjugated to other molecules such as lipids, proteins, sugars and the like. Peptides comprising D-amino acids generally have increased resistance to proteases, and thus have an extended serum half-life.
[0315] In addition, polypeptides comprising one or more peptide epitopes of the invention can be conjugated to other molecules such as lipids, proteins or sugars, or any other synthetic compounds, to increase their biological activity. For example, a T helper peptide can be conjugated to one or more palmitic acid chains at either the amino or the carboxyl termini.
Combinations of CTL Peptides with T Cell Priming Materials [0316] In some embodiments it may be desirable to include in the pharmaceutical compositions of the invention at least one component which primes cytotoxic T
lymphocytes. Lipids have been identified as agents capable of facilitating the priming in vitro CTL response against viral antigens. For example, palmitic acid residues can be attached to the e- and a-amino groups of a lysine residue and then linked to an immunogenic peptide. One or more linking moieties can be used such as Gly, Gly-Gly-, Ser, Ser-Ser, or the like. The lipidated peptide can then be administered directly in a micelle or particle, incorporated into a liposome, or emulsified in an adjuvant, e.g., incomplete Freund's adjuvant. A preferred immunogenic composition comprises palmitic acid attached to e- and a-amino groups of Lys via a linking moiety, e.g., Ser-Ser, added to the amino terminus of an immunogenic peptide.
[0317] In another embodiment of lipid-facilitated priming of CTL
responses, E.
coil lipoproteins, such as tripahnitoyl-S-glyceryl-cysteinyl-seryl-serine (P3CSS) can be used to prime CTL when covalently attached to an appropriate peptide.
(See, e.g., Deres, et al., Nature 342:561, 1989). Thus, peptides of the invention can be coupled to P3CSS, and the lipopeptide administered to an individual to specifically prime a CTL response to the target antigen. Moreover, because the induction of neutralizing antibodies can also be primed with P3CSS-conjugated epitopes, two such compositions can be combined to elicit both humoral and cell-mediated responses.
Vaccine Compositions Comprising Dendritic Cells Pulsed with CTL and/or HTL
Peptides [03181 An embodiment of a vaccine composition in accordance with the invention comprises ex vivo administration of a 'cocktail of epitope-bearing peptides to PBMC, or isolated DC therefrom, from the patient's blood. A
pharmaceutical to facilitate harvesting of DC can be used, such as ProgenipoietinTM (Monsanto, St. Louis, MO) or GM-CSF/IL-4. After pulsing the DC with peptides and prior to reinfusion into patients, the DC are washed to remove unbound peptides. In this embodiment, a vaccine comprises peptide-pulsed DCs which present the pulsed peptide epitopes in HLA molecules on their surfaces.
[03191 The DC can be pulsed ex vivo with a cocktail of peptides, some of which stimulate CTL responses to one or more antigens of interest, e.g., tumor associated antigens (TAA) such as HER2/neu, p53, MAGE 2, MAGE3, and/or carcinoembryonic antigen (CEA). Collectively, these TAA are associated with breast, colon and lung cancers. Optionally, a helper T cell (HTL) peptide such as PADRETM, can be included to facilitate the CTL response. Thus, a vaccine in accordance with the invention comprising epitopes from HER2/neu, p53, MAGE2, MAGE3, and carcinoembryonic antigen (CEA) is used to treat minimal or residual disease in patients with malignancies such as breast, colon or lung cancer; any malignancies that bear any of these TAAs can also be treated with the vaccine. A TAA vaccine can be used following debulking procedures such as surgery, radiation therapy or chemotherapy, whereupon the vaccine provides the benefit of increasing disease free survival and overall survival in the recipients.
[0320] Thus, in preferred embodiments, a vaccine of the invention is a product that treats a majority of patients across a number of different tumor types. A

vaccine comprising a plurality of epitopes, preferably supermotif-bearing epitopes, offers such an advantage.
Administration of Vaccines for Therapeutic or Prophylactic Purposes [0321] The polypeptides comprising one or more peptide epitopes of the present invention, including pharmaceutical and vaccine compositions thereof, are useful for administration to mammals, particularly humans, to treat and/or prevent disease. In one embodiment, vaccine compositions (peptide or nucleic acid) of the invention are administered to a patient who has a malignancy associated with expression of one or more TAAs, or to an individual susceptible to, or otherwise at risk for developing TAA-related disease. Upon administration an immune response is elicited against the TAAs, thereby enhancing the patient's own immune response capabilities. In therapeutic applications, peptide and/or nucleic acid compositions are administered to a patient in an amount sufficient to elicit an effective immune response to the TAA-expressing cells and to thereby cure, arrest or slow symptoms and/or complications. An amount adequate to accomplish this is defined as "therapeutically effective dose." Amounts effective for this use will depend on, e.g., the particular composition administered, the manner of administration, the stage and severity of the disease being treated, the weight and general state of health of the patient, and the judgment of the prescribing physician.
[0322] The vaccine compositions of the invention can be used purely as prophylactic agents. Generally the dosage for an initial prophylactic immunization generally occurs in a unit dosage range where the lower value is about 1, 5, 50, 500, or 1000 jig of peptide and the higher value is about 10,000;
20,000; 30,000; or 50,000 jig of peptide. Dosage values for a human typically range from about 500 jig to about 50,000 jig of peptide per 70 kilogram patient.
This is followed by boosting dosages of between about 1.0 ps to about 50,000 jig ofpeptide, administered at defined intervals from about four weeks to six months after the initial administration of vaccine. The immunogenicity of the vaccine may be assessed by measuring the specific activity of CTL and HTL obtained from a sample of the patient's blood.
[0323] As noted above, polypeptides comprising CTL and/or HTL epitopes of the invention induce immune responses when presented by HLA molecules and contacted with a CTL or HTL specific for an epitope comprised by the peptide.
The manner in which the peptide is contacted with the CTL or HTL is not critical to the invention. For instance, the peptide can be contacted with the CTL or HTL
either in vitro or in vivo. If the contacting occurs in vivo, peptide can be administered directly, or in other forms/vehicles, e.g., DNA vectors encoding one or more peptides, viral vectors encoding the peptide(s), liposomes, antigen presenting cells such as dendritic cells, and the like, as described herein.
[0324] Accordingly, for pharmaceutical compositions ofthe invention in the form of peptides or polypeptides, the peptides or polypeptides can be administered directly. Alternatively, the peptide/polypeptides can be administered indirectly presented on APCs, or as DNA encoding them. Furthermore, the polypeptides, peptide epitopes or DNA encoding them can be administered individually or as fusions of one or more peptide sequences.
[0325] For therapeutic use, administration should generally begin at the first diagnosis of TAA-related disease. This is followed by boosting doses at least until symptoms are substantially abated and for a period thereafter. In chronic disease states, loading doses followed by boosting doses may be required.
[0326] The dosage for an initial therapeutic immunization generally occurs in a unit dosage range where the lower value is about 1, 5, 50, 500, or 1,000 jig of peptide and the higher value is about 10,000; 20,000; 30,000; or 50,000 ps of peptide. Dosage values for a human typically range from about 500 jig to about 50,000 jig ofpeptide per 70 kilogram patient. Boosting dosages of between about 1.0 jig to about 50,000 lag of peptide, administered pursuant to a boosting regimen over weeks to months, can be administered depending upon the patient's response and condition. Patient response can be determined by measuring the specific activity of CTL and HTL obtained from the patient's blood.
[0327] In certain embodiments, polypeptides, peptides and compositions of the present invention are used in serious disease states. In such cases, as a result of the minimal amounts of extraneous substances and the relative nontoxic nature of the peptides, it is possible and may be desirable to administer substantial excesses of these peptide compositions relative to these stated dosage amounts.
[0328] For treatment of chronic disease, a representative dose is in the range disclosed above, namely where the lower value is about 1, 5, 50, 500, or 1,000 jig of peptide and the higher value is about 10,000; 20,000; 30,000; or 50,000 jig of peptide, preferably from about 500 lug to about 50,000 jig of peptide per 70 kilogram patient. Initial doses followed by boosting doses at established intervals, e.g., from four weeks to six months, may be required, possibly for a prolonged period of time to effectively immunize an individual. In the case of chronic disease, administration should continue until at least clinical symptoms or laboratory tests indicate that the disease has been eliminated or substantially abated, and for a follow-up period thereafter. The dosages, routes of administration, and dose schedules are adjusted in accordance with methodologies known in the art.
[0329] The pharmaceutical compositions for therapeutic treatment are intended for parenteral, topical, oral, intrathecal, or local administration.
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Claims (82)

THE EMBODIMENTS OF THE INVENTION FOR WHICH AN EXCLUSIVE
PROPERTY OR PRIVILEGE IS CLAIMED ARE DEFINED AS FOLLOWS:

1. An isolated polynucleotide comprising a sequence encoding (a) three complementarity determining regions (CDR) of the murine 1F2 monoclonal antibody variable light region (VL) encoded by SEQ ID NO: 148;
(b) three CDRs of the murine 1F2 monoclonal antibody variable heavy region (VH) encoded by SEQ ID NO: 149; or (c) (a) and (b).

2. The polynucleotide according to claim 1, wherein said polynucleotide comprises:
(a) positions 136 to 165 of SEQ ID NO:148;
(b) positions 211 to 231 of SEQ ID NO:148; and (c) positions 328 to 354 of SEQ ID NO:148.

3. The polynucleotide according to claim 1, wherein said polynucleotide comprises:
(a) positions 145 to 162 of SEQ ID NO:149;
(b) positions 205 to 252 of SEQ ID NO:149; and (c) positions 349 to 369 of SEQ ID NO:149.

4. An isolated polynucleotide comprising a sequence encoding (a) three complementarity determining regions (CDR) of the murine 1B3 monoclonal antibody variable light region (VL) encoded by SEQ ID NO: 150;
(b) three complementarity determining regions (CDR) of the murine 1B3 monoclonal antibody variable heavy region (VH) encoded by SEQ ID NO: 151; or (c) (a) and (b).

5. The polynucleotide according to claim 4, wherein said polynucleotide comprises:
(a) positions 130 to 162 of SEQ ID NO:150;
(b) positions 208 to 228 of SEQ ID NO:150; and (c) positions 325 to 351 of SEQ ID NO:150.

6. The polynucleotide according to claim 4, wherein said polynucleotide comprises:

(a) positions 145 to 159 of SEQ ID NO:151;
(b) positions 202 to 249 of SEQ ID NO:151; and (c) positions 346 to 387 of SEQ ID NO:151.

7. The polynucleotide according to claim 1, 2, 4 or 5, wherein said polynucleotide encodes a VL region that specifically binds C35.

8. The polynucleotide according to claim 7, further comprising a polynucleotide sequence encoding a human immunoglobulin light chain constant region or functional fragment thereof.

9. The polynucleotide according to claim 8, wherein said light chain constant region or functional fragment thereof is kappa.

10. The polynucleotide according to any one of claims 1, 3, 4 or 6, wherein said polynucleotide encodes a VH region that specifically binds C35.

11. The polynucleotide according to claim 10, further comprising a polynucleotide sequence encoding a human immunoglobulin heavy chain constant region or functional fragment thereof.

12. The polynucleotide according to claim 11, wherein said heavy chain constant region or functional fragment thereof is IgG.

13. The polynucleotide according to any one of claims 1 to 12, wherein said polynucleotide encodes an antibody or antigen binding fragment thereof that specifically binds C35, wherein said antibody or antigen binding fragment is humanized.

14. An isolated polynucleotide comprising a sequence having at least 80%
identity to SEQ ID NO:148 and encoding the same polypeptide as encoded by SEQ ID NO:148.

15. The polynucleotide according to claim 14 comprising a sequence having at least 85%
identity to SEQ ID NO:148.

16. The polynucleotide according to claim 15 comprising a sequence having at least 90%
identity to SEQ ID NO:148.

17. The polynucleotide according to claim 16 comprising a sequence having at least 95%
identity to SEQ ID NO:148.

18. The polynucleotide according to claim 17 comprising a sequence having at least 99%
identity to SEQ ID NO:148.

19. The polynucleotide according to claim 18 comprising the sequence of SEQ
ID
NO:148.

20. An isolated polynucleotide comprising a sequence having at least 80%
identity to SEQ ID NO:149 and encoding the same polypeptide as encoded by SEQ ID NO:149.

21. The polynucleotide according to claim 20 comprising a sequence having at least 85%
identity to SEQ ID NO:149.

22. The polynucleotide according to claim 21 comprising a sequence having at least 90%
identity to SEQ ID NO:149.

23. The polynucleotide according to claim 22 comprising a sequence having at least 95%
identity to SEQ ID NO:149.

24. The polynucleotide according to claim 23 comprising a sequence having at least 99%
identity to SEQ ID NO:149.

25. The polynucleotide according to claim 24 comprising the sequence of SEQ
ID
NO:149.

26. An isolated polynucleotide comprising a sequence having at least 80%
identity to SEQ ID NO:150 and encoding the same polypeptide as encoded by SEQ ID NO:150.

27. The polynucleotide according to claim 26 comprising a sequence having at least 85%
identity to SEQ ID NO:150.

28. The polynucleotide according to claim 27 comprising a sequence having at least 90%
identity to SEQ ID NO:150.

29. The polynucleotide according to claim 28 comprising a sequence having at least 95%
identity to SEQ ID NO:150.

30. The polynucleotide according to claim 29 comprising a sequence having at least 99%
identity to SEQ ID NO:150.

31. The polynucleotide according to claim 30 comprising the sequence of SEQ
ID
NO:150.

32. An isolated polynucleotide comprising a sequence having at least 80%
identity to SEQ ID NO:151 and encoding the same polypeptide as encoded by SEQ ID NO: 151.

33. The polynucleotide according to claim 32 comprising a sequence having at least 85%
identity to SEQ ID NO:151.

34. The polynucleotide according to claim 33 comprising a sequence having at least 90%
identity to SEQ ID NO:151.

35. The polynucleotide according to claim 34 comprising a sequence having at least 95%
identity to SEQ ID NO:151.

36. The polynucleotide according to claim 35 comprising a sequence having at least 99%
identity to SEQ ID NO:151.

37. The polynucleotide according to claim 36 comprising the sequence of SEQ
ID
NO:151.

38. An isolated polynucleotide comprising the sequence of SEQ ID NO:148 and the sequence of SEQ ID NO:149.

39. An isolated polynucleotide comprising the sequence of SEQ ID NO:150 and the sequence of SEQ ID NO:151.

40. The polynucleotide according to any one of claims 1 to 39, wherein said polynucleotide encodes an antibody or antigen binding fragment thereof that specifically binds C35.

41. An expression vector comprising the polynucleotide according to any one of claims 1 to 40.

42. A host cell comprising the expression vector according to claim 41.

43. A method of producing an isolated antibody or antigen binding fragment thereof that specifically binds C35 comprising culturing the host cell of claim 42, and recovering said antibody or antigen binding fragment thereof.

44. An antibody or antigen binding fragment thereof that specifically binds C35 produced by the method of claim 43.

45. An isolated polypeptide encoded by the polynucleotide according to any one of claims 1 to 40.

46. An isolated antibody or antigen binding fragment thereof which comprises at least an antigen binding region of a C35 antibody, wherein said antibody or antigen binding fragment thereof competes with said C35 antibody for binding to the same antigenic determinant and wherein said C35 antibody is the murine 1F2 monoclonal antibody comprising the variable light region (VL) encoded by SEQ ID NO: 148 and the variable heavy region (VH) encoded by SEQ ID NO: 149.

47. The antibody or antigen binding fragment thereof according to claim 46, wherein said antigen binding region comprises (a) a light variable region comprising the three light chain CDRs of the 1F2 murine monoclonal antibody and (b) a heavy variable region comprising the three heavy chain CDRs of the 1F2 murine monoclonal antibody.

48. The antibody or antigen binding fragment thereof according to claim 47, wherein said light variable region is encoded by SEQ ID NO: 148.

49. The antibody or antigen binding fragment thereof according to claim 47, wherein said heavy variable region is encoded by SEQ ID NO: 149.

50. The antibody or antigen binding fragment thereof according to any one of claims 46 to 49, wherein said antibody is humanized.

51. The antibody or antigen binding fragment thereof according to claim 46, wherein said antigen binding region comprises six complementarity determining regions (CDR) of said C35 antibody that areis grafted onto a framework region of a human antibody.

52. The antibody or antigen binding fragment thereof according to claim 51, wherein said six grafted CDRs have amino acid sequences encoded by positions 136 to 165 of SEQ ID
NO:148, positions 211 to 231 of SEQ ID NO:148, positions 328 to 354 of SEQ ID
NO:148, positions 145 to 162 of SEQ ID NO:149, positions 205 to 252 of SEQ ID NO:149, and positions 349 to 369 of SEQ ID NO:149.

53. The antibody or antigen binding fragment thereof according to any one of claims 46 to 52, wherein said antibody is an antigen recognizing fragment that specifically binds to C35.

54. The antibody according to claim 53, wherein said antigen recognizing fragment is a Fab fragment, a Fab' fragment, a F(ab')2 fragment, or a scFv fragment.

55. An isolated polynucleotide comprising a DNA segment encoding the antibody of any one of claims 46 to 54.

56. A method of preparing the antibody of any one of claims 46 to 54 comprising transfecting the polynucleotide of claim 55 into a host cell, expressing said antibody and recovering said antibody.

57. A composition comprising the antibody according to any one of claims 46 to 54 and a pharmaceutically acceptable carrier.

58. An isolated antibody or an antigen binding fragment thereof which comprises at least an antigen binding region of a C35 antibody, wherein said antibody or antigen binding fragment thereof competes with said C35 antibody for binding to the same antigenic determinant and wherein said C35 antibody is the murine 1B3 monoclonal antibody comprising the variable light region (VL) encoded by SEQ ID NO: 150 and the variable heavy region (VH) encoded by SEQ ID NO: 151.

59. The antibody or antigen binding fragment thereof according to claim 58, wherein said antigen binding region comprises (a) a light variable region comprising the three light chain CDRs of the 1B3 murine monoclonal antibody and (b) a heavy variable region comprising the three heavy chain CDRs of the 1B3 murine monoclonal.

60. The antibody or antigen binding fragment thereof according to claim 59, wherein said light variable region is encoded by SEQ ID NO: 150.

61. The antibody or antigen binding fragment thereof according to claim 59, wherein said heavy variable region is encoded by SEQ ID NO: 151.

62. The antibody or antigen binding fragment thereof according to any one of claims 58 to 61, wherein said antibody is humanized.

63. The antibody or antigen binding fragment thereof according to claim 58, wherein said antigen binding region comprises six complementarity determining regions (CDR) of said C35 antibody that are grafted onto a framework region of a human antibody.

64. The antibody or antigen binding fragment thereof according to claim 63, wherein said six grafted CDRs have amino acid sequences encoded by positions 130 to 162 of SEQ ID

NO:150, positions 208 to 228 of SEQ ID NO:150, positions 325 to 351 of SEQ ID
NO:150, positions 145 to 159 of SEQ ID NO:151, positions 202 to 249 of SEQ ID NO:151, and positions 346 to 387 of SEQ ID NO:151.

65. The antibody or antigen binding fragment thereof according to any one of claims 58 to 64, wherein said antibody is an antigen recognizing fragment that specifically binds to C35.

66. The antibody or antigen binding fragment thereof according to claim 65, wherein said antigen recognizing fragment is a Fab fragment, a Fab' fragment, a F(ab')2 fragment, or a scFv fragment.

67. An isolated polynucleotide comprising a DNA segment encoding the antibody of any one of claims 58 to 66.

68. A method of preparing the antibody or antigen binding fragment thereof of any one of claims 58 to 66 by a recombinant DNA technique, comprising transfecting the polynucleotide of claim 67 into a host cell, expressing said antibody or antigen binding fragment thereof and recovering said antibody or antigen binding fragment thereof.

69. A composition comprising the antibody or antigen binding fragment thereof according to any one of claims 58 to 66 and a pharmaceutically acceptable carrier.

70. An isolated polynucleotide encoding the antibody or antigen binding fragment thereof according to any one of claims 46 to 54 and 58 to 66.

71. A vector comprising the polynucleotide of claim 70.

72. A host cell comprising the vector of claim 71.

73. A composition comprising the antibody or antigen binding fragment thereof according to any one of claims 44, 46 to 54 and 58 to 66 and a pharmaceutically acceptable carrier.

74. The composition according to claim 73, wherein said composition further comprises a therapeutic agent.

75. The composition according to claim 74, wherein said therapeutic agent is an anti-tumor drug, a cytotoxin or a radioactive agent.

76. The composition according to claim 75, wherein said therapeutic agent is a cytotoxin.

77. The composition according to claim 76, wherein said cytotoxin is taxol, ricin, doxorubicin, cytochalasin B, gramicidin D, ethidium bromide, etoposide, tenoposide, colchicin, dihydroxy anthracin dione, 1-dehydrotestosterone or glucocorticoid.

78. The composition according to claim 77, wherein said cytotoxin is taxol.

79. The composition according to claim 74, wherein said therapeutic agent is conjugated to or complexed with said antibody or antigen binding fragment thereof.

80. A method of detecting the presence of C35 in a sample, said method comprising:
(a) contacting said sample with the antibody or antigen binding fragment thereof according to any one of claims 44, 46 to 54 and 58 to 66; and (b) assaying for binding of said antibody or antigen binding fragment thereof to C35.

81. The method according to claim 80, wherein said assaying step is performed by an assay selected from ELISA, radioimmunoassay, immunofluorescence or immunocytochemistry.

82. A diagnostic kit for detecting the presence of C35 in a sample, said kit comprising:
(a) the antibody or antigen binding fragment thereof according to any one of claims 44, 46 to 54 and 58 to 66; and (b) a conjugate comprising a specific binding partner for the C35 antibody and a label capable of producing a detectable signal.