INDIVIDUALIZED THERAPEUTIC ANTICANCER VACCINE

Abstract

The present invention relates to an individualized therapeutic anticancer vaccine, methods of treatment of cancer wherein such an anticancer vaccine is used as well as methods for producing the vaccine.

Claims

1. An individualized therapeutic anticancer vaccine comprising an immunologically effective amount of: (i) a polynucleotide comprising a nucleotide sequence encoding a targeting unit, a dimerization unit and an antigenic unit, wherein said antigenic unit comprises at least one patient-present shared antigen sequence or one or more parts thereof and wherein said antigenic unit further comprises one or more patient-specific antigen sequences or one or more parts thereof; or (ii) a polypeptide encoded by the polynucleotide as defined in (i); or (iii) a dimeric protein consisting of two polypeptides encoded by the polynucleotide as defined in (i); and a pharmaceutically acceptable carrier.

2. (canceled)

3. The vaccine according to claim 1, wherein said at least one patient-present shared antigen sequence is a sequence of a shared antigen selected from the group consisting of overexpressed cellular proteins, aberrantly expressed cellular proteins, cancer testis antigens, viral antigens, differentiation antigens, mutated oncogenes, mutated tumor suppressor genes, oncofetal antigens, shared fusion antigens, shared intron retention antigens, dark matter antigens, shared antigens caused by spliceosome mutations and shared antigens caused by frameshift mutations-, and/or wherein said at least one patient-present shared antigen sequence is a sequence of a shared antigen that is a human cellular protein, and/or wherein said at least one patient-present shared antigen sequence or one or more parts thereof are known to be immunogenic or are predicted to bind to the patient’s HLA class I and/or HLA class II alleles.

4-6. (canceled)

7. The vaccine according to claim 1, wherein the antigenic unit comprises sequences of several patient-present shared antigens or one or more parts thereof.

8. (canceled)

9. The vaccine according to claim 7, wherein the antigenic unit comprises several epitopes of several patient-present shared antigens, which epitopes are known to be immunogenic or are predicted to bind to the patient’s HLA class I and/or HLA class II alleles.

10. The vaccine according to claim 1, wherein the antigenic unit comprises one or more patient-present shared antigen sequences in full length, and/or wherein the antigenic unit comprises 1 to 30 parts of at least one patient-present shared antigen sequence, wherein the parts include multiple epitopes that are known to be immunogenic or are predicted to bind to the patient’s HLA class I and/or HLA class II alleles.

11-12. (canceled)

13. The vaccine according to claim 1, wherein the antigenic unit comprises 1 to 50 patient-present shared antigen sequences in the form of epitopes, wherein the epitopes are known to be immunogenic or predicted to bind to the patient’s HLA class I and/or HLA class II alleles.

14. (canceled)

15. The vaccine according to claim 13, wherein the epitopes have a length of from 7 to 30 amino acids.

16. The vaccine according to claim 1, wherein the antigenic unit comprises several patient-specific antigen sequences or one or more parts thereof, and/or wherein the antigenic unit comprises one or more patient-specific epitopes.

17-18. (canceled)

19. The vaccine according to claim 16, wherein antigenic unit comprises at least 5 patient-specific epitopes.

20-21. (canceled)

22. The vaccine according to claim 1, wherein the dimerization unit comprises a hinge region, and further comprises another domain that facilitates dimerization.

23-25. (canceled)

26. The vaccine according to claim 1, wherein the dimerization unit comprises hinge exons h1 and h4 connected through a linker to a CH3 domain of human IgG3.

27-28. (canceled)

29. The vaccine according to claim 1, wherein the targeting unit comprises or consists of soluble CD40 ligand, RANTES, MIP-1α, XCL1, XCL2, flagellin, anti-HLA-DP, anti-HLA-DR, anti-pan HLA class II or an antibody variable domain with specificity for anti-CD40, anti-TLR-2, anti-TLR-4 or anti-TLR-5 .

30. The vaccine according to claim 1, wherein said vaccine comprises a polynucleotide which is an RNA or DNA and further comprises a nucleotide sequence encoding a signal peptide.

31. The vaccine according to claim 30, wherein the polynucleotide is a DNA.

32. (canceled)

33. The vaccine according to claim 1, wherein the pharmaceutically acceptable carrier is selected from the group consisting of saline, buffered saline, PBS, dextrose, water, glycerol, ethanol, sterile isotonic aqueous buffers, and combinations thereof.

34. A method for preparing the individualized therapeutic anticancer vaccine of claim 1, said method comprises the steps of: a) identifying at least one patient-present shared antigen in the tumor tissue or body fluid of a patient b) determining the patient’s HLA class I and/or class II alleles c) predicting the immunogenicity of the identified at least one antigen or one or more parts thereof by their predicted binding to the patient’s HLA class I and/or II alleles d) selecting at least one antigen or one or more parts thereof based on their immunogenicity predicted in step c); e) preparing a polynucleotide sequence comprising an antigenic unit comprising a nucleotide sequence encoding the at least one antigen or one or more parts thereof selected in step d); f) cloning the polynucleotide sequence into an expression vector comprising nucleotide sequences encoding a targeting unit and a dimerization unit; and g) mixing the expression vector obtained in step f with a pharmaceutically acceptable carrier, and wherein said method further comprises: in step a) identifying one or more patient-specific antigens in the tumor tissue of the patient in step c) predicting the immunogenicity of the identified one or more patient-specific antigens or one or more parts thereof by their predicted binding to the patient’s HLA class I and/or II alleles in step d) selecting one or more patient-specific antigens or one or more parts thereof based on their immunogenicity predicted in step c); and wherein the polynucleotide sequence of step e) further comprises nucleotide sequences encoding the one or more patient-specific antigens or one or more parts thereof selected in step d).

35. (canceled)

36. A polynucleotide as defined in claim 1.

37. The polynucleotide according to claim 36 wherein the polynucleotide is comprised in a vector.

38-42. (canceled)

43. A method of treating a patient having cancer, the method comprising administering to the patient the vaccine according to claim 1, which has been prepared specifically for the patient.

44. (canceled)

45. The vaccine according to claim 1, wherein the targeting unit comprises or consists of MIP-1α.

Description

DESCRIPTION OF THE DRAWINGS

[0274] FIG. 1 shows the amino acid sequence of HPV16 E6 and all epitopes predicted to bind to the HLA class I alleles of patient 1. The two underlined sequences constitute HLA class I optimized sequences for inclusion into the antigenic unit of a therapeutic anticancer vaccine individualized for patient 1.

[0275] FIG. 2 shows the amino acid sequence of HPV16 E6 and all epitopes predicted to bind to the HLA class I and HLA class II alleles of patient 1. The sequence in the box constitutes an HLA class I/HLA class II optimized sequence for inclusion into the antigenic unit of a therapeutic anticancer vaccine individualized for patient 1.

[0276] FIG. 3 shows the amino acid sequence of HPV16 E7 and all epitopes predicted to bind to the HLA class I alleles of patient 1. The underlined sequence constitutes an HLA class I optimized sequence for inclusion into the antigenic unit of a therapeutic anticancer vaccine individualized for patient 1.

[0277] FIG. 4 shows the amino acid sequence of HPV16 E7 and all epitopes predicted to bind to the HLA class I and HLA class II alleles of patient 1. The sequences in the two boxes constitute HLA class I/HLA class II optimized sequences for inclusion into the antigenic unit of a therapeutic anticancer vaccine individualized for patient 1.

[0278] FIG. 5 shows the amino acid sequence of HPV16 E6 and all epitopes predicted to bind to the HLA class I alleles of patient 2. The underlined sequence constitutes an HLA class I optimized sequence for inclusion into the antigenic unit of a therapeutic anticancer vaccine individualized for patient 2.

[0279] FIG. 6 shows the amino acid sequence of HPV16 E6 and all epitopes predicted to bind to the HLA class I and HLA class II alleles of patient 2. The sequence in the box constitutes an HLA class I/HLA class II optimized sequence for inclusion into the antigenic unit of a therapeutic anticancer vaccine individualized for patient 2.

[0280] FIG. 7 shows the amino acid sequence of HPV16 E7 and all epitopes predicted to bind to the HLA class I alleles of patient 2. The underlined sequence constitutes an HLA class I optimized sequence for inclusion into the antigenic unit of a therapeutic anticancer vaccine individualized for patient 2.

[0281] FIG. 8 shows the amino acid sequence of HPV16 E7 and all epitopes predicted to bind to the HLA class I and HLA class II alleles of patient 2. The sequences in the two boxes constitute HLA class I/HLA class II optimized sequences for inclusion into the antigenic unit of a therapeutic anticancer vaccine individualized for patient 2.

[0282] FIG. 9 shows the immunogenicity of DNA vaccines (constructs) VB4097, VB4100 and VB4105 in mice vaccinated with these constructs by way of measuring the IFN-y immune responses from T cells (total T cell response), compared to the negative control VB1026.

[0283] FIG. 10 shows the immunogenicity of DNA vaccines (constructs) VB4100, VB4101 and VB4102 in mice vaccinated with these constructs by way of measuring the IFN-y immune responses from CD8+ T cells, compared to the negative control VB1026.

[0284] FIG. 11 shows the immunogenicity of DNA vaccines (constructs) VB4100 and VB4102 in mice vaccinated with these constructs by way of measuring the IFN-y immune responses from A) T cells (total T cell response), B) CD8+ T cells and C) CD4+ T cells, compared to the negative control VB1026.

[0285] FIG. 12 shows the immunogenicity of DNA vaccines (constructs) VB4118, VB4119, VB4121, VB4127, VB4128 and VB4130 in mice vaccinated with these constructs by way of measuring the IFN-y immune responses from T cells, compared to the negative control VB1026.

EXAMPLES

Example 1: Design of the Antigenic Unit Comprised in The Polynucleotide/Polypeptide/Dimeric Protein and Vaccine of the Invention

[0286] The antigenic unit may be designed with the following variations in the patient-present shared antigen sequence: [0287] A. Full-length sequence [0288] B. HLA-optimized sequences. The selected sequence for inclusion into the antigenic unit is optimized to cover the most immunogenic epitopes, i.e. those having a high binding affinity to the patient’s HLA I and/or HLA II molecules [0289] C. Epitopes with predicted binding to the patient’s HLA I and/or HLA II molecules. [0290] D. Combination of A+C [0291] E. Combination of B+C [0292] F. Combination of A+B

[0293] The patient-present shared antigen sequence above may for instance be HPV16. In cases D and F, which include a full-length sequence, case D may for instance be the combination of HPV16 E7 as a full-length sequence (A) and epitopes from HPV E6 (C) while case F may for instance be the combination of HPV16 E7 as a full-length sequence (A) and HLA-optimized sequences (B). The aforementioned example includes one single patient-present shared antigen but two different regions thereof. In another embodiment, cases D and F relate to the combination of the full-length sequence (A) of a first patient-present shared antigen, e.g. HPV16 E7 and epitopes (C) or HLA-optimized sequences (B) of a second patient-shared antigen, e.g. KRAS.

[0294] Thus, the antigenic unit may comprise A-F from one patient-present shared antigen or comprise A-F from several patient-present shared antigens.

[0295] Sequences A-F are arranged in the antigenic unit according to the methods provided herein. Different antigenic unit designs may be evaluated in animal models, e.g. as described in Example 3 to determine the optimal antigenic unit design. Breadth, strength and kinetics for the antigen-specific immunogenicity can be determined by IFN-gamma ELISPOT analysis. Anti-tumor efficacy can be tested in a tumor challenge experiment.

Example 2: Construction of a Polynucleotide According to the Invention

[0296] A polynucleotide according to the invention is designed and comprises the following units and components:

TABLE-US-00001 1: Native leader sequence for human LD78b Signal peptide 2: Full-length LD78b sequence Targeting unit 3: Human hinge-region 1 from IgG3 Dimerization unit 4: Human hinge region 4 from IgG3 5: Glycine-Serine linker 6: Human CH3 domain from IgG3 7: Glycine-Leucine linker Unit linker 8: Patient-present shared antigen sequence(s) Antigenic unit 9: Optionally: patient-specific antigen sequence(s)

EXAMPLE 3: Selecting Sequences of Patient-Present Shared Antigens for Inclusion Into the Antigenic Unit Comprised in the Polynucleotides/Polypeptides/Dimeric Peptides and Vaccines According to the Invention

[0297] Blood samples and tumor tissue samples of two patients — patient 1 and 2 — presenting with squamous cell carcinoma of the head and neck were obtained. The blood sample was analyzed for exome sequencing to characterize the exons in the healthy cells. The tumor tissue sample was analyzed for exome sequencing to characterize the exons in the tumor tissue and RNA-seq to evaluate RNA expression level of each gene. The presence of HPV16 shared antigen was identified using anti-HPV16 antibodies in an ELISA.

[0298] To find the most immunogenic sequences, each patients HLA class I and II alleles were determined by sequencing normal tissue (blood cells). The following HLA class I and II alleles were found:

TABLE-US-00002 Patient 1 - HLA class I alleles Patient 1 - HLA class II alleles HLA-A01:01 DRB1_0701 HLA-A24:02 DRB1_1301 HLA-B08:01 DRB3_0202 HLA-B40:01 DRB4_0101 HLA-C03:04 HLA-DQA10103-DQB10202 HLA-C07:01 HLA-DQA10103-DQB10603 HLA-DQA10201-DQB10202 HLA-DQA10201-DQB10603 HLA-A01:01 DRB1_0101 HLA-A24:02 DRB1_1301 HLA-B40:01 DRB3_0202 HLA-C03:04 HLA-DPA10103-DPB10401 HLA-DQA10101-DQB10501 HLA-DQA10101-DQB10603 HLA-DQA10103-DQB10501 HLA-DQA10103-DQB10603

[0299] HPVs have circular, double-stranded DNA genomes that are approximately 8 kb in size and encode eight genes, of which E6 and E7 have transforming properties. The viral E6 and E7 proteins are known to be involved in conversion of healthy cells into malignant cells. The abilities of HPV16 E6 and E7 proteins to associate with the tumor suppressors p53 and pRB, respectively, have been suggested as a mechanism by which these viral proteins induce tumors. Thus, the E6 and E7 sequences of HPV16 are known shared tumor antigens and were selected for finding sequences therein for inclusion into the antigenic unit. Prediction of binding to the patients’ HLA class I and HLA class II alleles was carried out using the NetMHCpan 4.0 software.

Patient 1

Hpv 16 E6

[0300] A total of 16 epitopes, each 9 amino acids long, were predicted to bind to HLA class I alleles of patient 1 and a total of 16 epitopes, each 9 amino acids long, were predicted to bind to HLA class II alleles of patient 1 (FIG. 2 and table 3)

[0301] Thus, an antigenic unit for inclusion into an individualized therapeutic anticancer vaccine for patient 1 is designed, which may comprise: [0302] A. The full-length HPV16 E6 sequence (151 amino acids) [0303] B. An HLA class I optimized sequence comprising some of the 16 epitopes, e.g. the 12 underlined epitopes shown on the left side of FIG. 1. This sequence contains 65 amino acids, i.e. 43% of the full-length sequence. [0304] C. Two HLA class I optimized sequences, wherein the first sequence comprises 12 of the 16 epitopes and the second sequence comprises 3 of the epitopes (FIG. 1). Thus, the two sequences combined include 15 of the 16 epitopes and contain 92 amino acids, i.e. 61% of the full-length sequence. [0305] D. An HLA class I/HLA class II optimized sequence, comprising 12 of the 16 HLA class I epitopes and 9 of the 16 HLA class II epitopes, thus including 21 of the 32 HLA class I/class II epitopes (FIG. 2, box). This sequence contains 65 amino acids, i.e. 43% of the full-length sequence.

[0306] Inclusion of C or D into the antigenic unit is preferred.

TABLE-US-00003 HPV E6 MFQDPQERPRKLPQLCTELQTTIHDIILECVYCKQQLLRREVYDFARRDLCIVYRDGNPYAV RDKCLKFYSKISEYRHYCYSLYGTTLEQQYNKPLCDLLIRCINRQKPLCPEEKQRHLDKKQR FHNIRGRWTGRCMSCCRSSRTRRETQL Epitope corresponding to amino acids in full length sequence Sequence of epitope 6-15 QERPRKLPQ 8-17 RPRKLPQLC 23-32 IHDIILECV 24-33 HDIILECVY 26-35 IILECVYCK 35-44 QQLLRREVY 42-51 VYDFARRDL 43-52 YDFARRDLC 44-53 DFARRDLCI 45-54 FARRDLCIV 60-69 YAVRDKCLK 62-71 VRDKCLKFY 81-90 YSLYGTTLE 125-134 FHNIRGRWT 131-140 RWTGRCMSC 143-152 SRTRRETQL 16-31 CTELQTTIHDIILEC 17-32 TELQTTIHDIILECV 18-33 ELQTTIHDIILECVY 19-34 LQTTIHDIILECVYC 20-35 QTTIHDIILECVYCK 34-49 KQQLLRREVYDFARR 49-64 DLCIVYRDGNPYAVR 50-65 LCIVYRDGNPYAVRD 51-66 CIVYRDGNPYAVRDK 78-93 HYCYSLYGTTLEQQY 95-110 PLCDLLIRCINRQKP 96-111 LCDLLIRCINRQKPL 97-112 CDLLIRCINRQKPLC 98-113 DLLIRCINRQKPLCP 99-114 LLIRCINRQKPLCPE 121-136 KKQRFHNIRGRWTGR

Hpv16 E7

[0307] A total of 9 epitopes, each 9 amino acids long, were predicted to bind to HLA class I alleles of patient 1 and a total of 12 epitopes, each 9 amino acids long, were predicted to bind to HLA class II alleles of patient 1 (FIG. 4 and table 4)

[0308] Thus, an antigenic unit for inclusion into an individualized therapeutic anticancer vaccine for patient 1 is designed, which may comprise: [0309] E. The full length HPV16 E7 sequence (98 amino acids) [0310] F. An HLA class I optimized sequence comprising some of the 9 epitopes, e.g. the underlined epitopes shown in FIG. 3. This sequence contains 56 amino acids, i.e. 57% of the full-length sequence [0311] G. Two HLA class I/HLA class II optimized sequences, wherein the first sequence comprises 2 of the 9 HLA class I epitopes and 7 of the 16 HLA class II epitopes and the second sequence comprises 4 of the 9 HLA class I epitopes and 5 of the 16 HLA class II epitopes (FIG. 4, boxes). Thus, the two sequences combined include 6 of the 9 HLA class I epitopes and all of the HLA class II epitopes. The two sequences combined contain 54 amino acids, i.e. 55% of the full-length sequence.

Inclusion of for G Into the Antigenic Unit is Preferred

[0312] Based on the above, an antigenic unit for inclusion into an individualized therapeutic anticancer vaccine for patient 1 is designed, which comprises at least one of C, D, F and G or all of C, D, F and G or any combination thereof between these two described extrema.

TABLE-US-00004 HPV E7 MHGDTPTLHEYMLDLQPETTDLYGYGQLNDSSEEEDEIDGPAGQAEPDRAHYNIVTFCCKC DSTLRLCVQSTHVDI RTLEDLLMGTLG IVCPICSQKP Epitope corresponding to amino acids in full length sequence Sequence of epitope 7-16 TLHEYMLDL 22-31 LYGYGQLND 38-47 IDGPAGQAE 48-57 DRAHYNIVT 55-64 VTFCCKCDS 66-75 RLCVQSTHV 72-81 THVDIRTLE 73-82 HVDIRTLED 85-94 GTLGIVCPI 4-19 DTPTLHEYMLDLQPE 5-20 TPTLHEYMLDLQPET 6-21 PTLHEYMLDLQPETT 7-22 TLHEYMLDLQPETTD 8-23 LHEYMLDLQPETTDL 9-24 HEYMLDLQPETTDLY 10-25 EYMLDLQPETTDLYG 70-85 QSTHVDIRTLEDLLM 71-86 STHVDIRTLEDLLMG 72-87 THVDIRTLEDLLMGT 73-88 HVDIRTLEDLLMGTL 74-89 VDIRTLEDLLMGTLG

Patient 2

Hpv16 E6

[0313] A total of 14 epitopes, each 9 amino acids long, were predicted to bind to HLA class I alleles of patient 2 and a total of 14 epitopes, each 9 amino acids long, were predicted to bind to HLA class II alleles of patient 2 (FIG. 6 and table 5)

[0314] Thus, an antigenic unit for inclusion into an individualized therapeutic anticancer vaccine for patient 2 is designed, which may comprise; [0315] H. The full-length HPV16 E6 sequence (151 amino acids) [0316] I. An HLA class I optimized sequence comprising some of the 14 epitopes, e.g. the 11 underlined epitopes shown in FIG. 5. This sequence contains 57 amino acids, i.e. 38% of the full-length sequence. [0317] J. An HLA class I/HLA class II optimized sequence, comprising 11 of the 14 HLA class I epitopes and 8 of the 14 HLA class II epitopes, thus including 19 of the 28 HLA class I/class II epitopes (FIG. 6, box). This sequence contains 59 amino acids, i.e. 39% of the full-length sequence.

[0318] Inclusion of J into the antigenic unit is preferred.

TABLE-US-00005 HPV E6 MFQDPQERPRKLPQLCTELQTTIHDIILECVYCKQQLLRREVYDFARRDLCIVYRDGNPYAV RDKCLKFYSKISEYRHYCYSLYGTTLEQQYNKPLCDLLIRCINRQKPLCPEEKQRHLDKKQR FHNIRGRWTGRCMSCCRSSRTRRETQL Epitope corresponding to amino acids in full length sequence Sequence of epitope 16-25 CTELQTTIH 42-51 VYDFARRDL 45-54 FARRDLCIV 52-61 IVYRDGNPY 54-63 YRDGNPYAV 68-77 KFYSKISEY 73-82 ISEYRHYCY 75-84 EYRHYCYSL 80-89 CYSLYGTTL 84-93 YGTTLEQQY 88-97 LEQQYNKPL 91-100 QYNKPLCDL 11-20 RHLDKKQRF 12-21 RFHNIRGRW 18-33 ELQTTIHDIILECVY 49-64 DLCIVYRDGNPYAVR 50-65 LCIVYRDGNPYAVRD 51-66 CIVYRDGNPYAVRDK 74-89 SEYRHYCYSLYGTTL 75-90 EYRHYCYSLYGTTLE 76-91 YRHYCYSLYGTTLEQ 77-92 RHYCYSLYGTTLEQQ 78-93 HYCYSLYGTTLEQQY 96-111 LCDLLIRCINRQKPL 97-112 CDLLIRCINRQKPLC 98-113 DLLIRCINRQKPLCP 99-114 LLIRCINRQKPLCPE 121-136 KKQRFHNIRGRWTGR

Hpv16 E7

[0319] A total of 10 epitopes, each 9 amino acids long, were predicted to bind to HLA class I alleles of patient 2 and a total of 11 epitopes, each 9 amino acids long, were predicted to bind to HLA class II alleles of patient 2 (FIG. 8 and table 6)

[0320] Thus, an antigenic unit for inclusion into an individualized therapeutic anticancer vaccine for patient 2 may be designed which comprises: [0321] A. The full length HPV16 E7 sequence (98 amino acids) [0322] B. An HLA class I optimized sequence comprising some of the 10 epitopes, e.g. the 6 underlined epitopes shown in FIG. 7. This sequence contains 26 amino acids, i.e. 27% of the full-length sequence. [0323] C. Two HLA class I/HLA class II optimized sequences, wherein the first sequence comprises 6 of the 10 HLA class I epitopes and 6 of the 11 HLA class II epitopes and the second sequence comprises 2 of the 10 HLA class I epitopes and 5 of the 11 HLA class II epitopes (FIG. 8, boxes). Thus, the two sequences combined include 8 of the 11 HLA class I epitopes and all of the HLA class II epitopes. The two sequences combined contain 45 amino acids, i.e. 45% of the full-length sequence.

[0324] Inclusion of L or M into the antigenic unit is preferred.

[0325] Based on the above, an antigenic unit for inclusion into an individualized therapeutic anticancer vaccine for patient 2 is designed, which comprises at least one of J, L and M or all of J, L and M or any combination thereof between these two described extrema.

TABLE-US-00006 HPV E7 MHGDTPTLHEYMLDLQPETTDLYGYGQLNDSSEEEDEIDGPAGQAEPDRAHYNIVTFCCKC DSTLRLCVQSTHVDIRTLEDLLMGTLGIVCPICSQKP Epitope corresponding to amino acids in full length sequence Sequence of epitope 3-12 GDTPTLHEY 7-16 TLHEYMLDL 9-18 HEYMLDLQP 15-24 LQPETTDLY 19-28 TTDLYGYGQ 20-29 TDLYGYGQL 44-53 QAEPDRAHY 49-58 RAHYNIVTF 71-80 STHVDIRTL 79-88 LEDLLMGTL 3-18 GDTPTLHEYMLDLQP 4-19 DTPTLHEYMLDLQPE 5-20 TPTLHEYMLDLQPET 6-21 PTLHEYMLDLQPETT 7-22 TLHEYMLDLQPETTD 8-23 LHEYMLDLQPETTDL 70-85 QSTHVDIRTLEDLLM 71-86 STHVDIRTLEDLLMG 72-87 THVDIRTLEDLLMGT 73-88 HVDIRTLEDLLMGTL 74-89 VDIRTLEDLLMGTLG

[0326] Comparing patient 1 and 2, i.e. FIGS. 2 and 6 and FIGS. 4 and 8, it is apparent that the optimal sequences for inclusion into a vaccine according to the invention differ considerably between the two patients.

Example 4: Comparing Vaccines Comprising Patient-Specific Antigens and Patient-Present Shared Antigens

[0327] To compare the efficacy of vaccines comprising only patient-specific antigen sequences with vaccines according to the invention comprising patient-present shared antigen sequences and optionally patient-specific antigen sequences, a mouse TC-2 tumor model is used.

[0328] Shared antigens and specific antigens present in the TC-2 tumor cell line are identified, processed and selected as described herein, i.e. shared antigen sequences for inclusion into the antigenic unit are selected based on their binding to MHC molecules while specific antigen sequences are selected based on additional parameters in an in silico predicted immunogenicity method. The shared antigen which is selected for inclusion into the antigenic unit is the viral antigen HPV16 and sequences encoding parts of the E6 and E7 proteins thereof were selected.

[0329] All selected antigen sequences are ordered from a commercial supplier, e.g. from Genscript (New Jersey, US) and cloned into the expression vector pUMVC4a, which comprises sequences encoding the LD78beta targeting unit and the hlgG3 dimerization unit.

[0330] The antigenic unit of vector 1 only comprises shared antigen sequences while the antigenic unit of vector 2 comprises both shared antigen sequences and specific antigen sequences.

[0331] To verify correct vaccibody formation, HEK293 cells are transfected with the vectors and vaccibody proteins in the supernatant are identified by Western blot and/or sandwich ELISA. The empty pUMVC4a vector is included as a negative control. Intact homodimeric protein formation is confirmed as follows: the proteins in the supernatant from transfected cells are detected in a Western blot by an anti-hMIP-1alpha antibody, in either the presence of reducing agents, which result in dimeric proteins being reduced to monomers, or absence of reducing agents.

[0332] A vaccine is prepared by mixing 20 .Math.g of the vector 1 and 2, respectively, with an aqueous buffer. The vaccine is injected intramuscularly in the tibial anterior muscle of the mouse followed by electroporation using TriGrid, Ichor, (US). At day 13, the mice are euthanized, and spleens are harvested.

[0333] The T cell responses are evaluated by IFN-gamma ELISpot. We observe that vaccines according to the present invention induce broader T cell responses that are higher compared to vaccines comprising only specific antigen sequences.

Example 5: Individual Therapeutic Anticancer DNA Vaccine According to the Invention

[0334] An individual therapeutic anticancer DNA vaccine according to the invention may be prepared by GMP manufacturing of the vector comprising the polynucleotide according to the invention according to regulatory authorities’ guidelines, and Fill & Finish of the DNA vaccine. The vector may be formulated by dissolving it in a sterile saline solution, such as PBS, at a concentration of 2-6 mg/ml. The vaccine may be administered either intradermally or intramuscularly with or without following electroporation or may alternatively be administered with a jet injector.

Example 6: Selecting Sequences of Patient-present Shared Antigens (and Patient-Specific Antigens) for Inclusion Into the Antigenic Unit Comprised in the Polynucleotides/Polypeptides/Dimeric Peptides and Vaccines According to the Invention

[0335] Blood samples and tumor tissue samples of three patients (patient 1, 2 and 3) presenting with squamous cell carcinoma of the head and neck were obtained. The blood sample was analyzed for exome sequencing to characterize the exons in the healthy cells. The tumor tissue sample was analyzed for exome sequencing to characterize the exons in the tumor tissue and RNA-seq to evaluate RNA expression level of each gene.

[0336] The presence of HPV16 shared antigen was identified using PCR. The ectopic expression of NY-ESO-1 was determined by anti-NY-ESO-1 antibodies in an ELISA. Patient-specific antigen sequences for each patient were identified as previously described in this application and in WO 2017/118695, which is included herein by reference.

[0337] NY-ESO-1 (also known as cancer/testis antigen 1B) is a protein belonging to the family of cancer-testis antigens (CTAs) that have been found to be re-expressed in a variety of malignant tumors at the mRNA and protein levels, while its normal expression in adult tissue is restricted to germ cells and placental cells. NY-ESO-1 expression has been reported in a wide range of tumor types.

[0338] For HPV, the E6 and E7 sequences of HPV16 are known shared tumor antigens and were selected for finding sequences therein for inclusion into the antigenic unit.

[0339] To find the most immunogenic sequences, each patients HLA class I and II alleles were determined by sequencing normal tissue (blood cells). Prediction of binding of HPV16 E6/E7 sequences and NY-ESO-1 sequences to the patients’ HLA class I and HLA class II alleles was carried out using the NetMHCpan 4.0 software. Included in the analysis were sequences from the IEDB database which are known from the literature to have elicited a positive T cell response (not matched to patients’ HLA alleles).

[0340] The HLA class I and II alleles which were found for patients 1, 2 and 3 are listed in the table below:

TABLE-US-00007 Patient 1 - HLA class I alleles Patient 1 - HLA class II alleles HLA-A02:01 DRB1_0701 HLA-A31:01 DRB1_1301 HLA-B37:01 DRB3_0202 HLA-B51:01 DRB4_0101 HLA-C06:02 HLA-DQA10103-DQB10202 HLA-C15:02 HLA-DQA10103-DQB10603 HLA-DQA10201-DQB10202 HLA-DQA10201-DQB10603 Patient 2 - HLA class I alleles Patient 2 - HLA class II alleles HLA-A01:01 DRB1_0101 HLA-A24:02 DRB1_1301 HLA-B08:01 DRB3_0202 HLA-B40:01 HLA-DPA10103-DPB10401 HLA-C03:04 HLA-DQA10101-DQB10501 HLA-C07:01 HLA-DQA10101-DQB10603 HLA-DQA10103-DQB10501 HLA-DQA10103-DQB10603 HLA-A02:01 DRB1_0112 HLA-B44:02 DRB1_0480 HLA-B57:01 DRB5_0202 HLA-C05:01 HLA-DPA10201-DPB10402 HLA-C06:02 HLA-DQA10102-DQB10301 HLA-DQA10101-DQB10603 HLA-DQA10201-DQB10501 HLA-DQA10103-DQB10603

Patient 1

[0341] TABLE-US-00008 Protein/Alleles/ IEDB epitopes Total number of epitopes Epitopes selected for inclusion into antigenic unit Remarks HPV16 E6 Full length E6 sequence selected, i.e. SEQ ID NO: 11 HLA I 25* 25 HLA II 16* 16 IEDB HLA I 26 26 IEDB HLA II 18 18 HPV 16 E7 Amino acid 69-98 of SEQ ID NO: 12 selected HLA I 16* 9 HLA II 12* 5 IEDB HLA I 20 7 IEDB HLA II 22 7 NY-ESO-1 Amino acid 80-101 of SEQ ID NO: 13 selected HLA I 26* HLA II 43* IEDB HLA I 7 IEDB HLA II 6 * total number of epitopes, each 9 amino acids long, predicted to bind to HLA class I and II alleles of patient 1, respectively.

[0342] An antigenic unit for inclusion into an individualized therapeutic anticancer vaccine for patient 1 was designed, which comprises SEQ ID NO: 14, comprising the sequences set forth in the right hand column of the table above in the following order: E7 |linker |NY-ESO-1 |linker |E6.

[0343] A second antigenic unit for inclusion into an individualized therapeutic anticancer vaccine for patient 1 was designed, which comprises SEQ ID NO: 15, comprising the sequences set forth in the right hand column of the table above and in addition 17 patient-specific antigen sequences. The most hydrophobic sequences were positioned substantially in the middle of the antigenic unit and the most hydrophilic sequence were positioned at the beginning and at end of the antigenic unit. Glycine-serine linkers were inserted between the sequences. The antigenic unit comprises the sequences in the following order, with T1D denoting the patient-specific antigen sequences:

[0344] T1D320 |linker |T1D814 |linker |T1D182 |linker | T1D689 |linker | E7 |linker |T1D339 |linker |T1D428 |linker |NY-ESO-1 |linker |T1D572 |linker |T1D359 |linker |T1D488 |linker | T1D554 |linker |T1D272 |linker |T1D210 |linker |T1D849 |linker | T1D4 |linker |T1D77 | linker |T1D717 |linker | T1D586 |linker | E6.

TABLE-US-00009 T1D320 HNEGDDQQGSRYSLIPQIQKVCEVVDG T1D814 SEQERMKSSLLKEHMLRKQAELESAQC T1D182 ELRENLLTYLPDSVTQLRRLEELDLGN T1D689 QEVFSSYKFNHLVRRLVLQREKHFHYL T1D339 IEKNADLCYLSTVHWSLILDAVSNNYI T1D428 LATAGEPYHDIRFKLMAVVPDRRIKYE T1D572 MLRLPTVFRQIRPVSRVLAPHLTR T1D359 IQIMENPFVQSMLWNPDLMRQLIMANP T1D488 LLRFLFLGLSALALPSRAQLQLHLPAN T1D554 MGKNPVRPPRAFPPVPSIDDIPLSR T1D272 GHVDFTIEVERALTVLDGAVLVLCAVG T1D210 EYKLMYGMLFSIRLFVSKMSPLDMKDG T1D849 SQGAGVESLDFRLYLRYEFLMLGIQPV T1D4 AAKTLVLCVSDIILLSANISETSSNKT T1D77 CKDLQLYLSNLANHIDRETGIGDVPLV T1D717 QSKHTEARELMYSAALLFFSHGQQNSA T1D586 MTLAESYAQYVYNLCNSLSIKVEES

Patient 2

[0345] TABLE-US-00010 Protein/Alleles/ IEDB epitopes Total number of epitopes Epitopes selected for inclusion into antigenic unit Remarks HPV16 E6 Amino acid 62-151 of SEQ ID NO: 11 selected HLA I 25* 15 HLA II 14* 10 IEDB HLA I 26 15 IEDB HLA II 18 13 HPV 16 E7 Amino acid 42-87 of SEQ ID NO: 12 selected HLA I 13* 5 HLA II 11* 5 IEDB HLA I 20 6 IEDB HLA II 22 11 NY-ESO-1 Amino acid 79-149 of SEQ ID NO: 13 selected HLA I 19* 19 HLA II 11* 11 IEDB HLA I 7 4 IEDB HLA II 6 3 * total number of epitopes, each 9 amino acids long, predicted to bind to HLA class I and II alleles of patient 2, respectively.

[0346] An antigenic unit for inclusion into an individualized therapeutic anticancer vaccine for patient 2 was designed, which comprises SEQ ID NO: 16, comprising the sequences set forth in the right hand column of the table above in the following order: E6 |linker |NY-ESO-1 |linker |E7.

[0347] A second antigenic unit for inclusion into an individualized therapeutic anticancer vaccine for patient 2 was designed, which comprises SEQ ID NO: 17, comprising the sequences set forth in the right hand column of the table above and in addition 17 patient-specific antigen sequences. The most hydrophobic sequences were positioned substantially in the middle of the antigenic unit and the most hydrophilic sequence were positioned at the beginning and at end of the antigenic unit. Glycine-serine linkers were inserted between the sequences. The antigenic unit comprises the sequences in the following order, with T1D denoting the patient-specific antigen sequences:

[0348] E6 |linker |T1D323 |linker |T1D506 |linker |T1D12 |linker |T1D315 |linker |T1D302 |linker | T1D700 |linker |NY-ESO-1 |linker |T1D535 |linker |T1D358 |linker |T1D670 |linker | T1D294 |linker |T1D336 |linker |T1D499 |linker |T1D425 |linker T1D491 |linker |T1D314 | linker |T1D430 |linker |E7 |linker |T1D582.

TABLE-US-00011 T11D323 KKVSKTRHTRETVFRRAKRRWAPIPCS T11D506 NLKHENILQFLTAQERKTELGKQYWLI T11D12 VQVFGLYFGEEFHETFDCPIK T11D315 GQAAASQAGGARGYARGAQLWPPGSDP T11D302 SIMQTWFTLFTPTDATSIVATTVMSNS Τ11D700 QENVRFVLVRSEALLPNAGPRSAEARV T11D535 DAFPNLKDFISRFQVMPPSSFLFDAPC T11D358 PSSLQVKPETPASEAVAVAAAAAPTTT T11D670 RFFHLADLFLSSSQLPAYLVAAFAKRL T11D294 LVFLWLHSLRRLFGCLYVSVFSNVMIH T11D336 WENAIAALFRRHIAVSWLIRATLSESE Τ11D499 RYIFVKSAGSRIEEGVLQFLVLLVAGR T11D425 QEIEWLPFRCIKCLKLSFSTAELLCMH T11D491 LVARCPPCLRLLRQARDFQAARYDRHD T11D314 ALGTPEDLDSYIDLSLESLNQMILELD T11D430 SQDMLSIMEKLEFLDFSYDLNLCGLTE T11D582 RREQYIPNEEFLHFDLLEDSKYRKIYS

Patient 3

[0349] TABLE-US-00012 Protein/Alleles/ IEDB epitopes Total number of epitopes Epitopes selected for inclusion into antigenic unit Remarks HPV16 E6 Amino acid 68-138 of SEQ ID NO: 11 selected HLA I 18* 6 HLA II 17* 8 IEDB HLA I 26 11 IEDB HLA II 18 6 HPV 16 E7 Amino acid 42-79 of SEQ ID NO: 12 selected HLA I 11* 3 HLA II 0* 0 IEDB HLA I 20 5 IEDB HLA II 22 8 NY-ESO-1 Amino acid 79–170 of SEQ ID NO: 13 selected HLA I 20* 20 HLA II 58* 42 IEDB HLA I 7 6 IEDB HLA II 6 6 * total number of epitopes, each 9 amino acids long, predicted to bind to HLA class I and II alleles of patient 3, respectively.

[0350] An antigenic unit for inclusion into an individualized therapeutic anticancer vaccine for patient 3 was designed, which comprises SEQ ID NO: 18, comprising the sequences set forth in the right hand column of the table above in the following order: NY-ESO-1 |linker | E7 |linker | E6.

[0351] A second antigenic unit for inclusion into an individualized therapeutic anticancer vaccine for patient 3 was designed, which comprises SEQ ID NO: 19, comprising the sequences set forth in the right hand column of the table above and in addition 17 patient-specific antigen sequences. The most hydrophobic sequences were positioned substantially in the middle of the antigenic unit and the most hydrophilic sequence were positioned at the beginning and at end of the antigenic unit. Glycine-serine linkers were inserted between the sequences. The antigenic unit comprises the sequences in the following order, with T1D denoting the patient-specific antigen sequences:

[0352] T1D223 |linker |T1D164 |linker |T1D56 |linker |T1D36 |linker |T1D129 |linker |T1D274 | linker |T1D62 |linker |T1D5 |linker |T1D144 |linker |T1D441 |linker |T1D368 |linker |NY-ESO-1 |linker |T1D234 |linker |T1D162 |linker |T1D39 |linker |T1D272 |linker |E7 |linker | T1D328 |linker |T1D188 |linker |E6.

TABLE-US-00013 T1D223 GPSYRSNSVSSLDLEGE T1D164 LKEEKENLQGLVTHQTYIIQELEKQLN T1D56 QRQKTAGKIFRAEVSTGQDAPRQAQAR T1D36 ILRPSTVNELESGEISYDEVGERIKDF T1D129 DTCINEDVESLRKMVQDLLAKLQEAKR T1D274 LREEGTKYKSFVQRARLVFREEGYLAF T1D62 HEGARPMRAIFLANGNVFTTGFSRMSE T1D5 IPDAAFIQAARRKCELARAQDDYISLD T1D144 MEIAKMRAGRRLWVHLIEKMFQPKNSK T1D441 MELVSFRDVAIEFSPEEW T1D368 TCVHEIPFHFDLMELLPQCQQLQMFFL T1D234 VRRVFITDDFHDMIPKYLNFVKGVVDS T1D162 NFHAHREKAPSLFCSRILNKAVYLFYG T1D39 DANKEGMFLFRAAHKLRQFLKMNSTGD T1D272 LLSLGWSVDVGRHSGWTGHVSTSWSIN T1D328 NTIFSLRKDLRQGKARRLRCMEEKEMF T1D188 NHFNDFEGDPAMTQFLEEFEKNLEDTK

Example 7: Immunogenicity of DNA Vaccines According to the Invention

Design of DNA Vaccines Comprising Shared Antigen Sequences And/or Neoepitopes

[0353] Five DNA vaccines (constructs) were designed comprising nucleotide sequences encoding the units/parts shown in table 14:

TABLE-US-00014 Unit/Part Function 1: Native leader sequence for human MIP1α (LD78β) Signal peptide 2: Full-length human MIP1α (LD78β) sequence Targeting unit 3: Human hinge-region 1 from IgG3 Dimerization unit 4: Human hinge region 4 from IgG3 5: Glycine-Serine linker 6: Human CH3 domain from IgG3 7: Glycine-Leucine linker Unit linker 8: Patient-present shared antigen sequences: Antigenic unit VB4100, VB4101, VB4102 9: Patient-specific antigen sequences (neoepitopes) VB4097 10: Patient-present shared antigen sequences and patient-specific antigen sequences (neoepitopes): VB4105

DNA Vaccine VB4097 Comprising 10 CT26 Neoepitopes

[0354] This construct was chosen as a model of an individualized DNA vaccine comprising patient-specific antigen sequences, i.e., neoepitopes.

[0355] Previously described exome sequencing and RNA sequencing of the mouse colon cancer cell line CT26 revealed hundreds to thousands of tumor-specific non-synonymous mutations. In silico methods were used to identify potential immunogenic sequences, i.e., neoepitopes, and 10 of them (table 15) were chosen for inclusion into the antigenic unit of VB4097. Each of the identified 10 neoepitopes is a peptide consisting of 27 amino acids. All but the terminal neoepitope were arranged in subunits, each subunit consisting of one neoepitope and one flexible glycine-serine linker (GGGGSGGGGS).

[0356] VB4097 consists of a DNA sequence encoding the polypeptide with amino acid sequence of SEQ ID NO: 20.

TABLE-US-00015 Neoantige n Gene Sequence Reactive T cell subtype C-pepM1 E2f8 VILPQAPSGPSYATYLQPAQAQMLT PP CD8+/CD4+ C-pepM6 UbqIn1 DTLSAMSNPRAMQVLLQIQQGLQT LAT CD4+ C-pepM8 Dhx35 EVIQTSKYYMRDVIAIESAWLLELAP H CD4+ C-pepM29 Anapc1 GSLFGSSRVQYVVNPAVKIVFLNID PS ND C-pepM31 Ptpn13 AEYGDYQPEVHGVPYFRLEHYLPA RVM CD4+ C-pepM43 Mtch1 KSWIHCWKYLSVQSQLFRGSSLLF RRV CD8+/CD4+ C-pepM89 Mmachc TLAFLVLSTPAMFNRALKPFLKSCH FQ CD4+ C-pep149 3110057O12R ik FVSPMAHYVPGIMAIESVVARFQFIV P CD8+ C-pepM171 Gdf11 LWVYLRPVPRPATIYLQILRLKPLTG E CD8+ C-pepM173 Top3a KIYEFDYHLYGQNITMIMTSVSGHLL A CD4+

DNA Vaccines VB4100, VB4101 and VB4102 Comprising NY-ESO-1 Sequences or Parts Thereof

[0357] These constructs were chosen as models of individualized DNA vaccines comprising patient-present shared antigen sequences.

[0358] Human New York esophageal squamous cell carcinoma 1 (NY-ESO-1) has been shown to be a highly immunogenic cancer testis antigen aberrantly expressed in several cancer types. NY-ESO-1 is not endogenously expressed in the CT26 cancer cell line, but using in silico methods, several immunogenic sequences were predicted to bind mouse MHC class I and II in BALB/c mice. The above-mentioned constructs were chosen as a model of an individualized DNA vaccine comprising patient-present shared antigen sequences.

[0359] Three NY-ESO-1 constructs were thus designed: [0360] VB4100 having an antigenic unit that comprises NY-ESO-1 in full length. VB4100 consists of a DNA sequence encoding the polypeptide with amino acid sequence of SEQ ID NO: 21. [0361] VB4101 having an antigenic unit that comprises amino acids 81-88 of the NY-ESO-1 sequence which is predicted to be an MHC class I antigen. VB4101 consists of a DNA sequence encoding the polypeptide with amino acid sequence of SEQ ID NO: 22. [0362] VB4102 having an antigenic unit that comprises amino acids 81-126 of the NY-ESO-1 sequence which contains several antigens that are predicted to be MHC class and class II antigens. VB4102 consists of a DNA sequence encoding the polypeptide with amino acid sequence of SEQ ID NO: 23.

[0363] The various NY-ESO-1 sequences comprised in the constructs are shown in table 16.

TABLE-US-00016 Construct NY-ESO-1 Sequence Length Reactive T cell subtype VB4100 Full length MQAEGRGTGGSTGDADGPGGPG IPDGPGGNAGGPGEAGATGGRG PRGAGAARASGPGGGAPRGPHG GAASGLNGCCRCGARGPESRLLE FYLAMPFATPMEAELARRSLAQD APPLPVPGVLLKEFTVSGNILTIRL TAADHRQLQLSISSCLQQLSLLMW ITQCFLPVFLAQPPSGQRR (SEQ ID NO: 13) 180 amino acids Both CD4+ and CD8+ VB4101 Amino acids 81-88 RGPESRLL 8 amino acids CD8+ VB4102 Amino Acids 81-126 RGPESRLLEFYLAMPFATPMEAEL ARRSLAQDAPPLPVPGVLLKEF 46 amino acids Both CD4+ and CD8+

DNA Vaccine VB4105 Comprising 10 CT26 Neoepitopes and the Full-Length Sequence of NY-ESO-1 Sequence

[0364] This construct was chosen as a model of an individualized DNA vaccine comprising patient-present shared antigen sequences and patient-specific antigen sequences (neoepitopes). It comprises an antigenic unit comprising the full-length sequence of NY-ESO-1 and the 10 CT26 neoepitopes shown in table 1, each of them separated from the following neoepitope or the NY-ESO-1 sequence by a flexible glycine-serine linker (GGGGSGGGGS). VB4105 consists of a DNA sequence encoding the polypeptide with amino acid sequence of SEQ ID NO: 24.

Negative Control VB1026

[0365] This construct is identical to the aforementioned constructs, but comprises neither a unit linker, nor an antigenic unit. It serves as a negative control. VB1026 consists of a DNA sequence encoding the polypeptide with amino acid sequence of SEQ ID NO: 25.

Construction of Expression Vectors Comprising the Constructs and Confirmation of Expression and Secretion of Intact Dimeric Proteins Encoded by the Constructs

[0366] The sequences of the antigenic units of all aforementioned constructs were ordered from Genscript (New Jersey, USA) and cloned into the expression vector pUMVC4a; a master plasmid comprising a nucleotide sequence encoding the signal peptide, targeting unit and dimerization unit described in table 14 above.

[0367] HEK293 cells (ATCC) were transiently transfected with the above-mentioned constructs. Briefly, 2×10.sup.5cells/well were plated in 24-well tissue culture plates with 10% FBS growth medium and transfected with 1 .Math.g of respective DNA plasmid using Lipofectamine® 2000 reagent under the conditions suggested by the manufacturer (Invitrogen, Thermo Fischer Scientific). The transfected cells were then maintained for up to 5 days at 37° C. with 5% CO.sub.2. Later the cell supernatants were harvested for characterization of the expression of the proteins encoded by the constructs by sandwich ELISA of the supernatant using antibodies specific for anti hIgG (CH3 domain), hMIP-1α and the respective antigenic units.

Assessment of Immunogenicity of the Constructs

[0368] Immunogenicity of the constructs was determined by way of measuring the T cell immune response elicited in vivo in mice vaccinated with the constructs.

[0369] Female, 6-week-old BALB/c mice were obtained from Janvier Labs (France). All animals were housed in the animal facility at the Radium Hospital (Oslo, Norway). All animal protocols were approved by the Norwegian Food Safety Authority (Oslo, Norway). 5 mice/group were used for the testing of the constructs comprising an antigenic unit, whereas 3 mice/group were used for the negative control.

[0370] 20 .Math.g of the construct was administered intramuscularly twice, i.e., on day 0 and on day 21, followed by electroporation. Spleens were collected on day 28.

[0371] Spleens were mashed in cell strainer to obtain a single cell suspension. For each construct tested, a portion of the single cell suspension was used to purify CD4+ and CD8+ T cells using Dynabeads® depletion. Total splenocytes, CD4 depleted splenocytes and CD8 depleted splenocytes were then tested for production of INF-γ in ELISpot assays. The 10 neoepitopes shown in table 15 were used to re-stimulate the splenocytes harvested from mice vaccinated with constructs VB4097 and VB4105, while the NY-ESO-1 peptides shown in table 17 below were used to re-stimulate the splenocytes harvested from mice vaccinated with the construct indicated in said table.

TABLE-US-00017 NY-ESO-1 peptides used for stimulation of splenocytes NY-ESO-1 peptide ID Sequence Length (amino acids) Used for constructs NY-ESO_Pep-1 MQAEGRGTGGSTGDA 15 VB4100, VB4105 NY-ESO_Pep-2 TGGSTGDADGPGGPG 15 VB4100, VB4105 NY-ESO_Pep-3 ADGPGGPGIPDGPGG 15 VB4100, VB4105 NY-ESO_Pep-4 GIPDGPGGNAGGPGE 15 VB4100, VB4105 NY-ESO_Pep-5 GNAGGPGEAGATGGR 15 VB4100, VB4105 NY-ESO_Pep-6 EAGATGGRGPRGAGA 15 VB4100, VB4105 NY-ESO_Pep-7 RGPRGAGAARASGPG 15 VB4100, VB4105 NY-ESO_Pep-8 AARASGPGGGAPRGP 15 VB4100, VB4105 NY-ESO_Pep-9 GGGAPRGPHGGAASG 15 VB4100, VB4105 NY-ESO_Pep-10 PHGGAASGLNGCCRC 15 VB4100, VB4105 NY-ESO_Pep-11 GLNGCCRCGARGPES 15 VB4100, VB4105 NY-ESO_Pep-12 CGARGPESRLLEFYL 15 VB4100, VB4102, VB4105 NY-ESO_Pep-13 RGPESRLLEFYLAMP 15 VB4100, VB4102, VB4105 NY-ESO_Pep-14 SRLLEFYLAMPFATP 15 VB4100, VB4102, VB4105 NY-ESO_Pep-15 LAMPFATPMEAELAR 15 VB4100, VB4102, VB4105 NY-ESO_Pep-16 PMEAELARRSLAQDA 15 VB4100, VB4102, VB4105 NY-ESO_Pep-17 RRSLAQDAPPLPVPG 15 VB4100, VB4102, VB4105 NY-ESO_Pep-18 DAPPLPVPGVLLKEF 15 VB4100, VB4102, VB4105 NY-ESO_Pep-19 APPLPVPGVLLKEFT 15 VB4100, VB4102, VB4105 NY-ESO_Pep-20 GVLLKEFTVSGNILT 15 VB4100, VB4105 NY-ESO_Pep-21 TVSGNILTIRLTAAD 15 VB4100, VB4105 NY-ESO_Pep-22 TIRLTAADHRQLQLS 15 VB4100, VB4105 NY-ESO_Pep-23 DHRQLQLSISSCLQQ 15 VB4100, VB4105 NY-ESO_Pep-24 SISSCLQQLSLLMWI 15 VB4100, VB4105 NY-ESO_Pep-25 QLSLLMWITQCFLPV 15 VB4100, VB4105 NY-ESO_Pep-26 ITQCFLPVFLAQPPS 15 VB4100, VB4105 NY-ESO_Pep-27 VFLAQPPSGQRR 12 VB4100, VB4105 NY-ESO_Pep-28 RGPESRLL 8 VB4100, VB4101, VB4102, VB4105

Comparison of Immunogenicity of Constructs VB4097, VB4100 and VB4105

[0372] Constructs VB4097 (10 neoepitopes), VB4100 (NY-ESO-1 full length), and VB4105 (10 neoepitopes and NY-ESO-1 full length) were compared for their ability to elicit T cell immune response against the peptides in tables 15 (VB4097 and VB4105) and 17 (VB4100 and VB4105).

[0373] As shown in FIG. 9, mice vaccinated with the negative control VB1026 showed low basal immunogenicity against both the neoepitopes and the NY-ESO-peptide sequences.

[0374] VB4097 and VB4105, both comprising the same 10 CT26 neoepitopes, induce a similar a total T cell response (INF-γ response) against the 10 neoepitopes (FIG. 9, grey bars), independent of whether the antigenic unit only comprises the 10 neoepitopes (VB4097) or comprises in addition the full-length sequence of NY-ESO-1 (VB4105).

[0375] Also, VB4100 and VB4105, both comprising the full-length sequence of NY-ESO-1 induce a similar a total T cell response (INF-γ response) against the peptides used for re-stimulation as shown in table 17 (FIG. 9, black bars), independent of whether the antigenic unit only comprises the full-length sequence of NY-ESO-1 (VB4100) or comprises in addition the 10 neoepitopes (VB4105).

[0376] For VB4105, the addition of the NY-ESO-1 full length sequence to the antigenic unit of VB4097 comprising the 10 neoepitopes did elicit a higher total T cell response compared to that elicited by vaccination with VB4097, due to the added immunogenicity against NY-ESO-1.

[0377] These results indicate that a vaccine according to the invention comprising a patient-present shared antigen is able to elicit an immune response which is similar to that of a vaccine comprising patient-specific antigens (neoepitopes). Moreover, these results indicate that there is a benefit in including both a patient-present shared antigen and patients-specific antigens (neoepitopes) into the antigenic unit.

Comparison of Immunogenicity of Constructs VB4100, VB4101 and VB4102

[0378] Using in silico methods, the epitope consisting of amino acids 81-88 of NY-ESO-1 was predicted to strongly bind to MHC class I to active CD8+ T cells, whereas the peptide consisting of amino acids 81 to 126 of NY-ESO-1 was predicted to contain several MHC class I and class II antigens. Therefore, constructs VB4100 (NY-ESO-1 full length), VB4101 (NY-ESO-1 amino acids 81-88) and VB4102 (NY-ESO-1 amino acids 81-126) were constructed and compared for their ability to elicit T cells immune response.

[0379] First, the three constructs were compared for their ability to elicit CD8+ T cell immune response against the predicted amino acid 81-88 region. The CD8+ T cells isolated from splenocytes of mice vaccinated with the constructs were re-stimulated with the amino acid 81-88 minimal epitope shown in table 16. As shown in FIG. 10, the experiments confirmed that the epitope consisting of amino acids 81-88 of NY-ESO-1 is indeed a strong CD8+ T cell epitope. Moreover, the immunogenicity for this region is independent of whether the epitope is the only sequence in the antigenic unit (VB4101) or the antigenic unit comprising a longer NY-ESO-1 sequence or the full NY-ESO-1 sequence.

[0380] Next, VB4102 was compared to VB4100 to assess whether the several predicted MHC class I and class II antigens in the amino acid sequence 81-126 of NY-ESO-1 elicit a similar response as the full-length NY-ESO-1 sequence. The CD4+ T cells and CD8+ T cells isolated from splenocytes of mice vaccinated with the constructs were re-stimulated with the peptides shown in table 17. As shown in FIG. 11, the amino acid sequence 81-126 of NY-ESO-1 elicits a stronger response than the NY-ESO-1 full length sequence, both from isolated CD8+ (FIG. 11B) and CD4+ T cells (FIG. 11C) and in terms of total T cell response (FIG. 11A).

[0381] These results suggest that by using in silico methods, short(er) sequences/epitopes of patient-present shared antigens can be identified that are predicted to elicit strong immune responses. By including such sequences/epitopes into the antigenic unit of the vaccine of the invention instead of longer sequences or the full-length sequence of a patient-present shared antigen, there is space left in the antigenic unit for including sequences of other patient-present shared antigens and/or patient-specific antigens/neoepitopes. This will enhance the chance that the patient receiving such an individual anticancer vaccine will show a strong immune response to the vaccine.

Example 8: Immunogenicity of DNA Vaccines According to the Invention Design of DNA Vaccines Comprising Shared Antigen Sequences and/or Neoepitopes

[0382] Six DNA vaccines (constructs) were designed comprising nucleotide sequences encoding the units/parts shown in table 18:

TABLE-US-00018 Unit/Part Function 1: Native leader sequence for human MIP1α (LD78β) Signal peptide 2: Full-length human MIP1α (LD78β) sequence Targeting unit 3: Human hinge-region 1 from IgG3 Dimerization unit 4: Human hinge region 4 from IgG3 5: Glycine-Serine linker 6: Human CH3 domain from IgG3 7: Glycine-Leucine linker Unit linker 8: Patient-present shared antigen sequences: Antigenic unit VB4119, VB4127 9: Patient-specific antigen sequences (neoepitopes) VB4118 10: Patient-present shared antigen sequences and patient-specific antigen sequences (neoepitopes): VB4121, VB4128, VB4130

DNA Vaccine VB4118 Comprising 10 B16 Neoepitopes

[0383] This construct was chosen as a model of an individualized DNA vaccine comprising patient-specific antigen sequences, i.e. neoepitopes.

[0384] Previously described exome sequencing and RNA sequencing of the mouse melanoma cell line B16.F10 revealed hundreds to thousands of tumor-specific non-synonymous mutations. In silico methods were used to identify potential immunogenic sequences, i.e. neoepitopes, and 10 of them (table 19) were chosen for inclusion into the antigenic unit of VB4118. Each of the identified 10 neoepitopes is a peptide consisting of 27 amino acids. All but the terminal neoepitope were arranged in subunits, each subunit consisting of one neoepitope and one flexible glycine-serine linker (GGGGSGGGGS).

[0385] VB4118 consists of a DNA sequence encoding the polypeptide with amino acid sequence of SEQ ID NO: 26.

TABLE-US-00019 Neoantige n Gene Sequence Reactive T cell subtype B-pepM2 ObsI1 REGVELCPGNKYEMRRHGTTHSLVIHD CD8+ B-pepM7 Atp11a SSPDEVALVEGVQSLGFTYLRLKDNYM CD8+ B-pepM36 Pcmtd 1 KNILAVSFAPLVQLSKNDNGTPDSVGL CD4+ B-pepM78 KIhI26 PAADRWEPRAPMRAPRVLHAMLGAAG R CD4+ B-pepM79 B3galt 6 VLSADLVHYLRLSLEYLRAWHSEDVSL CD4+ B-pepM82 Mta1 LEAVLRYLETHPRLPKPDPVKSSSSVL CD4+ B-pepM83 Ddit4I LKDFFFSRGRFSSALKRTLILSSGFRL CD4+ B-pepM84 Wdr3 AGKDHKIKQWDADTFEHIQTLEGHHQE CD8+ B-pepM85 Rfx3 PATIEMAIETLQKFDGLSTHRSSLLNS ND B-pepM86 Map1s YLPGGGAGHLDQNVFLRVRALCYVISG CD8+

DNA Vaccines VB4119 Comprising a TRP-2 Sequence and VB4127 Comprising Frameshift Antigens

[0386] These constructs were chosen as models of individualized DNA vaccine comprising patient-present shared antigen sequences.

[0387] Tyrosinase related protein 2 (TRP-2) is a melanocyte lineage normal differentiation protein. This shared antigen (differentiation antigen) is known to induce tumor rejection of B16 melanoma cells in C57BL/6 mice in vivo. In the literature, the 9 amino acid long MHC class I epitope shown in table 20 (amino acids 180-188 of TRP-2) has been identified as the immunogenic sequence responsible for the anti-tumor effect of TRP-2.

[0388] VB4119 consists of a DNA sequence encoding the polypeptide with amino acid sequence of SEQ ID NO: 27

TABLE-US-00020 Construct TRP-2 epitope Sequence Length Reactive T cell subtype VB4119 Amino acids 180-188 SVYDFFVWL 9 amino acids CD8+

[0389] Frameshift mutations are DNA mutations that arise when indels (insertions or deletions) of nucleotides lead to a shift in the DNA reading frame. Consequently, the entire DNA sequence following the indel will be read incorrectly and the resulting protein will be altered. Frameshift mutations arising in tumor cells generate novel peptide sequences that could be highly immunogenic, and, moreover, identical frameshift antigens could arise across different patients, and thus represent promising targets for shared antigen cancer vaccines (see for instance Ballhausen et al., Nat. Commun. 11, 2020, 1-13). The three frameshift antigens shown in table 21 were identified as immunogenic by in silico methods and are encoded in the VB4127 construct. In the antigenic unit, they are separated from each other by a flexible glycine-serine linker (GGGGSGGGGS).

[0390] VB4127 consists of a DNA sequence encoding the polypeptide with amino acid sequence of SEQ ID NO: 28.

TABLE-US-00021 Frameshift antigen Gene Sequence Reactive T cell subtype B-pepM108 Maz YISDHMKVHSPSPCL CD4+ B-pepM115-M122 Dync1h1 EGWQTCWGRSRKHWGSTWN GSARLSPGSTLWVMRICLRSL GIARTWLSCRSTSRKCSPAFP ASS CD4+/CD8+ B-pepM141-M142 Prtg LFRLLPSGPKVNDGHRSRRW HSLDLYPHLCSHLDLPKQSQE VIRL CD8+

DNA Vaccines Comprising 10 B16 Neoepitopes and the TRP-2 Epitope (VB4121) or 10 B16 Neoepitopes and the 3 Frameshift Antigens (VB4128) or 10 B16 Neoepitopes, the TRP-2 Epitope and the 3 Frameshift Antigens (VB4130)

[0391] These constructs were chosen as models of individualized DNA vaccines comprising patient-present shared antigen sequences and patient-specific antigen sequences (neoepitopes). Each construct comprises an antigenic unit comprising the 10 B16 neoepitopes shown in table 19, each of them separated from the following neoepitope or the shared antigen sequence by a flexible glycine-serine linker (GGGGSGGGGS).The same linker was used to separate the shared antigens sequences in constructs comprising several of such sequences. VB4121 consists of a DNA sequence encoding the polypeptide with amino acid sequence of SEQ ID NO: 29; VB4128 consists of a DNA sequence encoding the polypeptide with amino acid sequence of SEQ ID NO: 30 and VB4130 consists of a DNA sequence encoding the polypeptide with amino acid sequence of SEQ ID NO: 31.

[0392] The construct VB1026, described in Example 7, was used as a negative control.

Construction of Expression Vectors Comprising the Constructs and Confirmation of Expression and Secretion of Intact Dimeric Proteins Encoded by the Constructs

[0393] The expression vectors comprising the aforementioned constructs were constructed as described in Example 7. Expression and secretion of intact dimeric proteins encoded by the constructs was confirmed as described in Example 7.

Assessment of Immunogenicity of the Constructs

[0394] Immunogenicity of the constructs was determined by way of measuring the T cell immune response elicited in vivo in mice vaccinated with the constructs.

[0395] Female, 6-week-old C57BL/6 mice were obtained from Janvier Labs (France). All animals were housed in the animal facility at the Radium Hospital (Oslo, Norway). All animal protocols were approved by the Norwegian Food Safety Authority (Oslo, Norway). 5 mice/group were used for the testing of the constructs comprising an antigenic unit, whereas 3 mice/group were used for the negative control.

[0396] 20 .Math.g of the construct was administered intramuscularly on day 0 followed by electroporation. The spleens from the vaccinated mice were collected on day 14. The collected spleens were processed and ELISpot assays were carried out as described in Example 7.

[0397] The 10 neoepitopes shown in table 19 were used to re-stimulate the splenocytes harvested from mice vaccinated with the constructs VB4118, VB4121, VB4128, and VB4130. The TRP-2 peptide in table 20 was used to re-stimulate the splenocytes harvested from mice vaccinated with the constructs VB4119, VB4121, and VB4130. The frameshift peptides shown in table 22 below were used to re-stimulate the splenocytes harvested from mice vaccinated with the constructs VB4127, VB4128, and VB4130.

TABLE-US-00022 Gene Frameshift peptide ID Sequence Length Maz B-pepM108 YISDHMKVHSPSPCL Dync1h1 B-pepM115 EGWQTCWGRSRKHWG 15 amino acids Dync1h1 B-pepM116 GRSRKHWGSTWNGSA 15 amino acids Dync1h1 B-pepM117 GSTWNGSARLSPGST 15 amino acids Dync1h1 B-pepM118 ARLSPGSTLWVMRIC 15 amino acids Dync1h1 B-pepM119 TLWVMRICLRSLGIA 15 amino acids Dync1h1 B-pepM120 CLRSLGIARTWLSCR 15 amino acids Dync1h1 B-pepM121 ARTWLSCRSTSRKCS 15 amino acids Dync1h1 B-pepM122 RSTSRKCSPAFPASS 15 amino acids Prtg B-pepM141 LFRLLPSGPKVNDGHRSRRWHSLDLYP 27 amino acids Prtg B-pepM142 SRRWHSLDLYPHLCSHLDLPKQSQEVIRL 29 amino acids

Comparison of Immunogenicity of Constructs VB4118, VB4119, VB4121, VB4127, VB4128, and VB4130

[0398] Constructs VB4118 (10 neoepitopes), VB4119 (TRP-2), VB4121 (10 neoepitopes and TRP-2), VB4127 (3 frameshift antigens), VB4128 (10 neoantigens and 3 frameshift antigens), and VB4130 (10 neoepitopes, 3 frameshift antigens and TRP-2) were compared for their ability to elicit T cell immune response against the peptides in tables 19, 20, and 22, as applicable.

[0399] As shown in FIG. 12, mice vaccinated with the negative control VB1026 showed low basal immunogenicity against all the peptides tested.

[0400] Both the model construct of a vaccine comprising only patient-specific antigens (VB4118) and the model constructs of vaccines comprising only patient-present shared antigen(s) (VB4119 and VB4127) elicit an immune response in the vaccinated mice.

[0401] VB4118, VB4121, VB4128, and VB4130, all comprising the same 10 B16 neoepitopes, induce a similar total T cell response (INF-γ response) against the 10 neoepitopes (FIG. 12, black bars), independent of whether the antigenic unit only comprises the 10 neoepitopes (VB4118) or comprises in addition the TRP-2 epitope (VB4121), the 3 frameshift antigens (VB4128) or the TRP-2 epitope plus the 3 frameshift antigens (VB4130).

[0402] Further, FIG. 12 shows that adding the shared antigens to a neoepitope construct leads to a broader and increased total T cell response with the highest total T cell response observed for the construct encoding 10 neoepitopes, 3 frameshift antigens and the TRP-2 epitope (VB4130).

[0403] Also the results of this study indicate that a vaccine according to the invention comprising a patient-present shared antigen can elicit an immune response which is similar to that of a vaccine comprising patient-specific antigens (neoepitopes). Moreover, these results indicate that there is the benefit of an increased broader and total T cell response by including different types of shared antigens and patients-specific antigens (neoepitopes) into the antigenic unit.

SEQUENCES

[0404] SEQ ID NO: 1 C-C motif chemokine 3-like 1 precursor including signal peptide (aa 1-23) and mature peptide (hMIP1α/LD78-beta, aa 24-93):

TABLE-US-00023 MQVSTAALAVLLCTMALCNQVLSAPLAADTPTACCFSYTSRQIPQNFIAD YFETSSQCSKPSVIFLTKRGRQVCADPSEEWVQKYVSDLELSA

TABLE-US-00024 SEQ ID NO: 2 MQVSTAALAVLLCTMALCNQVLS | APLAADTPTACCFSYTSRQIPQNFIAD YFETSSQCSKPSVIFLTKRGRQVCADPSEEWVQKYVSDLELSA | ELKTPLG DTTHT I EPKSCDTPPPCPRCP | GGGSSGGGSG | GQPREPQVYTLPPSREEMTK NQVSLTCLVKGFYPSDIAVEWESSGQPENNYNTTPPMLDSDGSFFLYSKL TVDKSRWQQGNIFSCSVMHEALHNRFTQKSLSLSPGK | GLGGL |

[0405] SEQ ID NO: 3 Linker, amino acid sequence: GLSGL

[0406] SEQ ID NO: 4 Linker, amino acid sequence: GLGGL

[0407] SEQ ID NO: 5 Hinge region 1 (human IgG3 UH hinge), 12 amino acids: ELKTPLGDTTHT

[0408] SEQ ID NO: 6 Hinge region 4 (human IgG3, MH hinge, 15 amino acids): EPKSCDTPPPCPRCP

[0409] SEQ ID NO: 7 Gly-Ser Linker:

TABLE-US-00025 GGGSSGGGSG

[0410] SEQ ID NO: 8 hCH3 IgG3, amino acid sequence:

TABLE-US-00026 GQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESSGQPENNYNTTPPMLDSDG SFFLYSKL TVDKSRWQQG NIFSCSVM H EALH N RFTQKSLSLSPGK

SEQ ID NO: 9 Signal peptide

TABLE-US-00027 MNFGLRLIFLVLTLKGVQC

[0411] SEQ ID NO: 10 Signal peptide

TABLE-US-00028 MDAMKRGLCCVLLLCGAVFVSP

[0412] SEQ ID NO: 11: HPV16 E6

TABLE-US-00029 MFQDPQERPRKLPQLCTELQTTIHDIILECVYCKQQLLRREVYDFARRDL CIVYRDGNPYAVRDKCLKFYSKISEYRHYCYSLYGTTLEQQYNKPLCDLL IRCINRQKPLCPEEKQRHLDKKQRFHNIRGRWTGRCMSCCRSSRTRRETQ L

[0413] SEQ ID NO: 12: HPV16 E7

TABLE-US-00030 MHGDTPTLHEYMLDLQPETTDLYGYGQLNDSSEEEDEIDGPAGQAEPDRA HYNIVTFCCKCDSTLRLCVQSTHVDIRTLEDLLMGTLGIVCPICSQKP

[0414] SEQ ID NO: 13: NY-ESO-1

TABLE-US-00031 MQAEGRGTGGSTGDADGPGGPGIPDGPGGNAGGPGEAGATGGRGPRGAGA ARASGPGGGAPRGPHGGAASGLNGCCRCGARGPESRLLEFYLAMPFATPM EAELARRSLAQDAPPLPVPGVLLKEFTVSGNILTIRLTAADHRQLQLSIS SCLQQLSLLMWITQCFLPVFLAQPPSGQRR

[0415] SEQ ID NO: 14 (302 amino acids) Antigenic unit comprising antigen sequences in the following order: E7 |linker | NY-ESO-1 | linker |E6.

TABLE-US-00032 VQSTHVDIRTLEDLLMGTLGIVCPICSQKPGGGGSGGGGSARGPESRLLE FYLAMPFATPMEAELARRSLAQDAPPLPVPGVLLKEFTVSGNILTIRLTA ADHRQLQLSISSCLQQLSLLMWITQCFLPVFLAQPPSGQRRSGGGGSGGG GMFQDPQERPRKLPQLCTELQTTIHDIILECVYCKQQLLRREVYDFARRD LCIVYRDGNPYAVRDKCLKFYSKISEYRHYCYSLYGTTLEQQYNKPLCDL LIRCINRQKPLCPEEKQRHLDKKQRFHNIRGRWTGRCMSCCRSSRTRRET QL

[0416] SEQ ID NO: 15 (924 amino acids) Antigenic unit comprising antigen sequences in the following order: T1D320 |linker |T1D814 | linker |T1D182 |linker |T1D689 |linker |E7 |linker |T1D339 |linker |T1D428 |linker |NY-ESO-1 |linker |T1D572 |linker |T1D359 |linker |T1D488 |linker |T1D554 |linker |T1D272 | linker |T1D210 |linker |T1D849 |linker |T1D4 |linker |T1D77 |linker |T1D717 |linker | T1D586 |linker |E6.

TABLE-US-00033 HNEGDDQQGSRYSLIPQIQKVCEVVDGGSSGGGSSGGSEQERMKSSLLKE HMLRKQAELESAQCSGGGGSGGGGELRENLLTYLPDSVTQLRRLEELDLG NGGGGSGGGGSQEVFSSYKFNHLVRRLVLQREKHFHYLSGGGGSGGGGVQ STHVDIRTLEDLLMGTLGIVCPICSQKPGGGGSGGGGSIEKNADLCYLST VHWSLILDAVSNNYIGGGGSGGGGSLATAGEPYHDIRFKLMAVVPDRRIK YEGGGGSGGGGSARGPESRLLEFYLAMPFATPMEAELARRSLAQDAPPLP VPGVLLKEFTVSGNILTIRLTAADHRQLQLSISSCLQQLSLLMWITQCFL PVFLAQPPSGQRRSGGGGSGGGGMLRLPTVFRQIRPVSRVLAPHLTRSGG GGSGGGGIQIMENPFVQSMLWNPDLMRQLIMANPGGGGSGGGGSLLRFLF LGLSALALPSRAQLQLHLPANGGGGSGGGGSMGKNPVRPPRAFPPVPSID DIPLSRGSGGGGSGGGGHVDFTIEVERALTVLDGAVLVLCAVGSSGGGSS GGGEYKLMYGMLFSIRLFVSKMSPLDMKDGGSSGGGSSGGSQGAGVESLD FRLYLRYEFLMLGIQPVGGGGSGGGGSAAKTLVLCVSDIILLSANISETS SNKTSGGGGSGGGGCKDLQLYLSNLANHIDRETGIGDVPLVGGGGSGGGG SQSKHTEARELMYSAALLFFSHGQQNSAGGGGSGGGGSMTLAESYAQYVY NLCNSLSIKVEESSGGGGSGGGGMFQDPQERPRKLPQLCTELQTTIHDII LECVYCKQQLLRREVYDFARRDLCIVYRDGNPYAVRDKCLKFYSKISEYR HYCYSLYGTTLEQQYNKPLCDLLIRCINRQKPLCPEEKQRHLDKKQRFHN IRGRWTGRCMSCCRSSRTRRETQL

[0417] SEQ ID NO: 16 (227 amino acids) Antigenic unit comprising antigen sequences in the following order: E6 | linker | NY-ESO-1 | linker | E7

TABLE-US-00034 VRDKCLKFYSKISEYRHYCYSLYGTTLEQQYNKPLCDLLIRCINRQKPLC PEEKQRHLDKKQRFHNIRGRWTGRCMSCCRSSRTRRETQLSGGGGSGGGG GARGPESRLLEFYLAMPFATPMEAELARRSLAQDAPPLPVPGVLLKEFTV SGNILTIRLTAADHRQLQLSIGGGGSGGGGSAGQAEPDRAHYNIVTFCCK CDSTLRLCVQSTHVDIRTLEDLLMGTL

[0418] SEQ ID NO: 17 (850 amino acids) Antigenic unit comprising antigen sequences in the following order: E6 | linker | T1D323 | linker | T1D506 | linker | T1D12 | linker | T1D315 | linker | T1D302 | linker | T1D700 | linker | NY-ESO-1 | linker | T1 D535 | linker | T1D358 | linker | T1D670 | linker | T1 D294 | linker | T1 D336 | linker | T1D499 | linker | T1D425 | linker T1D491 | linker | T1D314 | linker | T1D430 | linker | E7 | linker | T1D582.

TABLE-US-00035 VRDKCLKFYSKISEYRHYCYSLYGTTLEQQYNKPLCDLLIRCINRQKPLC PEEKQRHLDKKQRFHNIRGRWTGRCMSCCRSSRTRRETQLSGGGGSGGGG KKVSKTRHTRETVFRRAKRRWAPIPCSSGGGGSGGGGNLKHENILQFLTA QERKTELGKQYWLIGGGGSGGGGSVQVFGLYFGEEFHETFDCPIKGSGGG GSGGGGQAAASQAGGARGYARGAQLWPPGSDPGGGGSGGGGSSIMQTWFT LFTPTDATSIVATTVMSNSSGGGGSGGGGQENVRFVLVRSEALLPNAGPR SAEARVGGGGSGGGGSGARGPESRLLEFYLAMPFATPMEAELARRSLAQD APPLPVPGVLLKEFTVSGNILTIRLTAADHRQLQLSIGGGGSGGGGSDAF PNLKDFISRFQVMPPSSFLFDAPCSGGGGSGGGGPSSLQVKPETPASEAV AVAAAAAPTTTSGGGGSGGGGRFFHLADLFLSSSQLPAYLVAAFAKRLSG GGGSGGGGLVFLWLHSLRRLFGCLYVSVFSNVMIHSGGGGSGGGGWENAI AALFRRHIAVSWLIRATLSESEGGGGSGGGGSRYIFVKSAGSRIEEGVLQ FLVLLVAGRSGGGGSGGGGQEIEWLPFRCIKCLKLSFSTAELLCMHGGGG SGGGGSLVARCPPCLRLLRQARDFQAARYDRHDGGGGSGGGGSALGTPED LDSYIDLSLESLNQMILELDGGGGSGGGGSSQDMLSIMEKLEFLDFSYDL NLCGLTEGGGGSGGGGSAGQAEPDRAHYNIVTFCCKCDSTLRLCVQSTHV DIRTLEDLLMGTLGGGSGGGGSGRREQYIPNEEFLHFDLLEDSKYRKIYS

[0419] SEQ ID NO: 18 (221 amino acids): Antigenic unit comprising antigen sequences in the following order: NY-ESO-1 | linker | E7 | linker | E6.

TABLE-US-00036 GARGPESRLLEFYLAMPFATPMEAELARRSLAQDAPPLPVPGVLLKEFTV SGNILTIRLTAADHRQLQLSISSCLQQLSLLMWITQCFLPVFGGGGSGGG GSAGQAEPDRAHYNIVTFCCKCDSTLRLCVQSTHVDIRTLSGGGGSGGGG KFYSKISEYRHYCYSLYGTTLEQQYNKPLCDLLIRCINRQKPLCPEEKQR HLDKKQRFHNIRGRWTGRCMS

[0420] SEQ ID NO: 19 (831 amino acids) Antigenic unit comprising antigen sequences in the following order: T1D223 | linker | T1D164 | linker | T1D56 | linker | T1D36 | linker | T1D129 | linker | T1D274 | linker | T1D62 | linker | T1D5 | linker | T1D144 | linker | T1D441 | linker | T1D368 | linker | NY-ESO-1 | linker | T1D234 | linker | T1D162 | linker | T1D39 | linker | T1D272 | linker | E7 | linker | T1D328 | linker | T1D188 | linker | E6.

TABLE-US-00037 GPSYRSNSVSSLDLEGEGGGGSGGGGSLKEEKENLQGLVTHQTYIIQELE KQLNGGGGSGGGGSQRQKTAGKIFRAEVSTGQDAPRQAQARGGGSSGGGS SILRPSTVNELESGEISYDEVGERIKDFGGGGSGGGGSDTCINEDVESLR KMVQDLLAKLQEAKRSGGGGSGGGGLREEGTKYKSFVQRARLVFREEGYL AFGGGGSGGGGSHEGARPMRAIFLANGNVFTTGFSRMSEGGGGSGGGGSI PDAAFIQAARRKCELARAQDDYISLDGGGGSGGGGSMEIAKMRAGRRLWV HLIEKMFQPKNSKGGGGSGGGGSMELVSFRDVAIEFSPEEWGGGGSGGGG STCVHEIPFHFDLMELLPQCQQLQMFFLGGGSGGGGSGGARGPESRLLEF YLAMPFATPMEAELARRSLAQDAPPLPVPGVLLKEFTVSGNILTIRLTAA DHRQLQLSISSCLQQLSLLMWITQCFLPVFGGGGSGGGGSVRRVFITDDF HDMIPKYLNFVKGVVDSSGGGGSGGGGNFHAHREKAPSLFCSRILNKAVY LFYGSSGGGSSGGGDANKEGMFLFRAAHKLRQFLKMNSTGDGGGGSGGGG SLLSLGWSVDVGRHSGWTGHVSTSWSINGGGGSGGGGSAGQAEPDRAHYN IVTFCCKCDSTLRLCVQSTHVDIRTLSGGGGSGGGGNTIFSLRKDLRQGK ARRLRCMEEKEMFGGGGSGGGGSNHFNDFEGDPAMTQFLEEFEKNLEDTK GGGGSGGGGSKFYSKISEYRHYCYSLYGTTLEQQYNKPLCDLLIRCINRQ KPLCPEEKQRHLDKKQRFHNIRGRWTGRCMS

[0421] SEQ ID NO: 20 Amino acid sequence of VB4097

TABLE-US-00038 MQVSTAALAVLLCTMALCNQVLSAPLAADTPTACCFSYTSRQIPQNFIAD YFETSSQCSKPSVIFLTKRGRQVCADPSEEWVQKYVSDLELSAELKTPLG DTTHTEPKSCDTPPPCPRCPGGGSSGGGSGGQPREPQVYTLPPSREEMTK NQVSLTCLVKGFYPSDIAVEWESSGQPENNYNTTPPMLDSDGSFFLYSKL TVDKSRWQQGNIFSCSVMHEALHNRFTQKSLSLSPGKGLGGLKIYEFDYH LYGQNITMIMTSVSGHLLAGGGGSGGGGSAEYGDYQPEVHGVPYFRLEHY LPARVMGGGGSGGGGSGSLFGSSRVQYVVNPAVKIVFLNIDPSGGGGSGG GGSLWVYLRPVPRPATIYLQILRLKPLTGEGGGGSGGGGSTLAFLVLSTP AMFNRALKPFLKSCHFQGGGGSGGGGSFVSPMAHYVPGIMAIESVWARFQ FIVPGGGGSGGGGSVILPQAPSGPSYATYLQPAQAQMLTPPGGGGSGGGG SEVIQTSKYYMRDVIAIESAWLLELAPHGGGGSGGGGSDTLSAMSNPRAM QVLLQIQQGLQTLATGGGGSGGGGSKSWIHCWKYLSVQSQLFRGSSLLFR RV

[0422] SEQ ID NO: 21 Amino acid sequence of VB4100

TABLE-US-00039 MQVSTAALAVLLCTMALCNQVLSAPLAADTPTACCFSYTSRQIPQNFIAD YFETSSQCSKPSVIFLTKRGRQVCADPSEEWVQKYVSDLELSAELKTPLG DTTHTEPKSCDTPPPCPRCPGGGSSGGGSGGQPREPQVYTLPPSREEMTK NQVSLTCLVKGFYPSDIAVEWESSGQPENNYNTTPPMLDSDGSFFLYSKL TVDKSRWQQGNIFSCSVMHEALHNRFTQKSLSLSPGKGLGGLMQAEGRGT GGSTGDADGPGGPGIPDGPGGNAGGPGEAGATGGRGPRGAGAARASGPGG GAPRGPHGGAASGLNGCCRCGARGPESRLLEFYLAMPFATPMEAELARRS LAQDAPPLPVPGVLLKEFTVSGNILTIRLTAADHRQLQLSISSCLQQLSL LMWITQCFLPVFLAQPPSGQRR

[0423] SEQ ID NO: 22 Amino acid sequence of VB4101

TABLE-US-00040 MQVSTAALAVLLCTMALCNQVLSAPLAADTPTACCFSYTSRQIPQNFIAD YFETSSQCSKPSVIFLTKRGRQVCADPSEEWVQKYVSDLELSAELKTPLG DTTHTEPKSCDTPPPCPRCPGGGSSGGGSGGQPREPQVYTLPPSREEMTK NQVSLTCLVKGFYPSDIAVEWESSGQPENNYNTTPPMLDSDGSFFLYSKL TVDKSRWQQGNIFSCSVMHEALHNRFTQKSLSLSPGKGLGGLRGPESRLL

[0424] SEQ ID NO: 23 Amino acid sequence of VB4102

TABLE-US-00041 MQVSTAALAVLLCTMALCNQVLSAPLAADTPTACCFSYTSRQIPQNFIAD YFETSSQCSKPSVIFLTKRGRQVCADPSEEWVQKYVSDLELSAELKTPLG DTTHTEPKSCDTPPPCPRCPGGGSSGGGSGGQPREPQVYTLPPSREEMTK NQVSLTCLVKGFYPSDIAVEWESSGQPENNYNTTPPMLDSDGSFFLYSKL TVDKSRWQQGNIFSCSVMHEALHNRFTQKSLSLSPGKGLGGLRGPESRLL EFYLAMPFATPMEAELARRSLAQDAPPLPVPGVLLKEF

[0425] SEQ ID NO: 24 Amino acid sequence of VB4105

TABLE-US-00042 MQVSTAALAVLLCTMALCNQVLSAPLAADTPTACCFSYTSRQIPQNFIAD YFETSSQCSKPSVIFLTKRGRQVCADPSEEWVQKYVSDLELSAELKTPLG DTTHTEPKSCDTPPPCPRCPGGGSSGGGSGGQPREPQVYTLPPSREEMTK NQVSLTCLVKGFYPSDIAVEWESSGQPENNYNTTPPMLDSDGSFFLYSKL TVDKSRWQQGNIFSCSVMHEALHNRFTQKSLSLSPGKGLGGLKIYEFDYH LYGQNITMIMTSVSGHLLAGGGGSGGGGSAEYGDYQPEVHGVPYFRLEHY LPARVMGGGGSGGGGSGSLFGSSRVQYVVNPAVKIVFLNIDPSGGGGSGG GGSLWVYLRPVPRPATIYLQILRLKPLTGEGGGGSGGGGSTLAFLVLSTP AMFNRALKPFLKSCHFQGGGGSGGGGSFVSPMAHYVPGIMAIESVVARFQ FIVPGGGGSGGGGSVILPQAPSGPSYATYLQPAQAQMLTPPGGGGSGGGG SEVIQTSKYYMRDVIAIESAWLLELAPHGGGGSGGGGSDTLSAMSNPRAM QVLLQIQQGLQTLATGGGGSGGGGSKSWIHCWKYLSVQSQLFRGSSLLFR RVGGGGSGGGGSMQAEGRGTGGSTGDADGPGGPGIPDGPGGNAGGPGEAG ATGGRGPRGAGAARASGPGGGAPRGPHGGAASGLNGCCRCGARGPESRLL EFYLAMPFATPMEAELARRSLAQDAPPLPVPGVLLKEFTVSGNILTIRLT AADHRQLQLSISSCLQQLSLLMWITQCFLPVFLAQPPSGQRR

[0426] SEQ ID NO: 25 Amino acid sequence of VB1026

TABLE-US-00043 MQVSTAALAVLLCTMALCNQVLSAPLAADTPTACCFSYTSRQIPQNFIAD YFETSSQCSKPSVIFLTKRGRQVCADPSEEWVQKYVSDLELSAELKTPLG DTTHTEPKSCDTPPPCPRCPGGGSSGGGSGGQPREPQVYTLPPSREEMTK NQVSLTCLVKGFYPSDIAVEWESSGQPENNYNTTPPMLDSDGSFFLYSKL TVDKSRWQQGNIFSCSVMHEALHNRFTQKSLSLSPGK

[0427] SEQ ID NO: 26 Amino acid sequence of VB4118

TABLE-US-00044 MQVSTAALAVLLCTMALCNQVLSAPLAADTPTACCFSYTSRQIPQNFIAD YFETSSQCSKPSVIFLTKRGRQVCADPSEEWVQKYVSDLELSAELKTPLG DTTHTEPKSCDTPPPCPRCPGGGSSGGGSGGQPREPQVYTLPPSREEMTK NQVSLTCLVKGFYPSDIAVEWESSGQPENNYNTTPPMLDSDGSFFLYSKL TVDKSRWQQGNIFSCSVMHEALHNRFTQKSLSLSPGKGLGGLREGVELCP GNKYEMRRHGTTHSLVIHDGGGGSGGGGSSSPDEVALVEGVQSLGFTYLR LKDNYMGGGGSGGGGSLKDFFFSRGRFSSALKRTLILSSGFRLGGGGSGG GGSKNILAVSFAPLVQLSKNDNGTPDSVGLGGGGSGGGGSLEAVLRYLET HPRLPKPDPVKSSSSVLGGGGSGGGGSPAADRWEPRAPMRAPRVLHAMLG AAGRGGGGSGGGGSVLSADLVHYLRLSLEYLRAWHSEDVSLGGGGSGGGG SYLPGGGAGHLDQNVFLRVRALCYVISGGGGGSGGGGSPATIEMAIETLQ KFDGLSTHRSSLLNSGGGGSGGGGSAGKDHKIKQWDADTFEHIQTLEGHH QE

[0428] SEQ ID NO: 27 Amino acid sequence of VB4119

TABLE-US-00045 MQVSTAALAVLLCTMALCNQVLSAPLAADTPTACCFSYTSRQIPQNFIAD YFETSSQCSKPSVIFLTKRGRQVCADPSEEWVQKYVSDLELSAELKTPLG DTTHTEPKSCDTPPPCPRCPGGGSSGGGSGGQPREPQVYTLPPSREEMTK NQVSLTCLVKGFYPSDIAVEWESSGQPENNYNTTPPMLDSDGSFFLYSKL TVDKSRWQQGNIFSCSVMHEALHNRFTQKSLSLSPGKGLGGLSVYDFFVW L

[0429] SEQ ID NO: 28 Amino acid sequence of VB4127

TABLE-US-00046 MQVSTAALAVLLCTMALCNQVLSAPLAADTPTACCFSYTSRQIPQNFIAD YFETSSQCSKPSVIFLTKRGRQVCADPSEEWVQKYVSDLELSAELKTPLG DTTHTEPKSCDTPPPCPRCPGGGSSGGGSGGQPREPQVYTLPPSREEMTK NQVSLTCLVKGFYPSDIAVEWESSGQPENNYNTTPPMLDSDGSFFLYSKL TVDKSRWQQGNIFSCSVMHEALHNRFTQKSLSLSPGKGLGGLYISDHMKV HSPSPCLGGGGSGGGGSEGWQTCWGRSRKHWGSTWNGSARLSPGSTLWVM RICLRSLGIARTWLSCRSTSRKCSPAFPASSGGGGGSGGGGSLFRLLPSG PKVNDGHRSRRWHSLDLYPHLCSHLDLPKQSQEVIRL

[0430] SEQ ID NO: 29 Amino acid sequence of VB4121

TABLE-US-00047 MQVSTAALAVLLCTMALCNQVLSAPLAADTPTACCFSYTSRQIPQNFIAD YFETSSQCSKPSVIFLTKRGRQVCADPSEEWVQKYVSDLELSAELKTPLG DTTHTEPKSCDTPPPCPRCPGGGSSGGGSGGQPREPQVYTLPPSREEMTK NQVSLTCLVKGFYPSDIAVEWESSGQPENNYNTTPPMLDSDGSFFLYSKL TVDKSRWQQGNIFSCSVMHEALHNRFTQKSLSLSPGKGLGGLREGVELCP GNKYEMRRHGTTHSLVIHDGGGGSGGGGSSSPDEVALVEGVQSLGFTYLR LKDNYMGGGGSGGGGSLKDFFFSRGRFSSALKRTLILSSGFRLGGGGSGG GGSKNILAVSFAPLVQLSKNDNGTPDSVGLGGGGSGGGGSLEAVLRYLET HPRLPKPDPVKSSSSVLGGGGSGGGGSPAADRWEPRAPMRAPRVLHAMLG AAGRGGGGSGGGGSVLSADLVHYLRLSLEYLRAWHSEDVSLGGGGSGGGG SYLPGGGAGHLDQNVFLRVRALCYVISGGGGGSGGGGSPATIEMAIETLQ KFDGLSTHRSSLLNSGGGGSGGGGSAGKDHKIKQWDADTFEHIQTLEGHH QEGGGGSGGGGSSVYDFFVWL

[0431] SEQ ID NO: 30 Amino acid sequence of VB4128

TABLE-US-00048 MQVSTAALAVLLCTMALCNQVLSAPLAADTPTACCFSYTSRQIPQNFIAD YFETSSQCSKPSVIFLTKRGRQVCADPSEEWVQKYVSDLELSAELKTPLG DTTHTEPKSCDTPPPCPRCPGGGSSGGGSGGQPREPQVYTLPPSREEMTK NQVSLTCLVKGFYPSDIAVEWESSGQPENNYNTTPPMLDSDGSFFLYSKL TVDKSRWQQGNIFSCSVMHEALHNRFTQKSLSLSPGKGLGGLYISDHMKV HSPSPCLGGGGSGGGGSEGWQTCWGRSRKHWGSTWNGSARLSPGSTLWVM RICLRSLGIARTWLSCRSTSRKCSPAFPASSGGGGGSGGGGSLFRLLPSG PKVNDGHRSRRWHSLDLYPHLCSHLDLPKQSQEVIRLGGGGSGGGGSREG VELCPGNKYEMRRHGTTHSLVIHDGGGGSGGGGSSSPDEVALVEGVQSLG FTYLRLKDNYMGGGGSGGGGSLKDFFFSRGRFSSALKRTLILSSGFRLGG GGSGGGGSKNILAVSFAPLVQLSKNDNGTPDSVGLGGGGSGGGGSLEAVL RYLETHPRLPKPDPVKSSSSVLGGGGSGGGGSPAADRWEPRAPMRAPRVL HAMLGAAGRGGGGSGGGGSVLSADLVHYLRLSLEYLRAWHSEDVSLGGGG SGGGGSYLPGGGAGHLDQNVFLRVRALCYVISGGGGGSGGGGSPATIEMA IETLQKFDGLSTHRSSLLNSGGGGSGGGGSAGKDHKIKQWDADTFEHIQT LEGHHQE

[0432] SEQ ID NO: 31 Amino acid sequence of VB4130

TABLE-US-00049 MQVSTAALAVLLCTMALCNQVLSAPLAADTPTACCFSYTSRQIPQNFIAD YFETSSQCSKPSVIFLTKRGRQVCADPSEEWVQKYVSDLELSAELKTPLG DTTHTEPKSCDTPPPCPRCPGGGSSGGGSGGQPREPQVYTLPPSREEMTK NQVSLTCLVKGFYPSDIAVEWESSGQPENNYNTTPPMLDSDGSFFLYSKL TVDKSRWQQGNIFSCSVMHEALHNRFTQKSLSLSPGKGLGGLYISDHMKV HSPSPCLGGGGSGGGGSEGWQTCWGRSRKHWGSTWNGSARLSPGSTLWVM RICLRSLGIARTWLSCRSTSRKCSPAFPASSGGGGGSGGGGSLFRLLPSG PKVNDGHRSRRWHSLDLYPHLCSHLDLPKQSQEVIRLGGGGSGGGGSREG VELCPGNKYEMRRHGTTHSLVIHDGGGGSGGGGSSSPDEVALVEGVQSLG FTYLRLKDNYMGGGGSGGGGSLKDFFFSRGRFSSALKRTLILSSGFRLGG GGSGGGGSKNILAVSFAPLVQLSKNDNGTPDSVGLGGGGSGGGGSLEAVL RYLETHPRLPKPDPVKSSSSVLGGGGSGGGGSPAADRWEPRAPMRAPRVL HAMLGAAGRGGGGSGGGGSVLSADLVHYLRLSLEYLRAWHSEDVSLGGGG SGGGGSYLPGGGAGHLDQNVFLRVRALCYVISGGGGGSGGGGSPATIEMA IETLQKFDGLSTHRSSLLNSGGGGSGGGGSAGKDHKIKQWDADTFEHIQT LEGHHQEGGGGSGGGGSSVYDFFVWLGGGGSGGGGS

[0433] SEQ ID NO: 32 Amino acids 6-15 of HPV E6

TABLE-US-00050 QERPRKLPQ

[0434] SEQ ID NO: 33 Amino acids 8-17 of HPV E6

TABLE-US-00051 RPRKLPQLC

[0435] SEQ ID NO: 34 Amino acids 23-32 of HPV E6

TABLE-US-00052 IHDIILECV

[0436] SEQ ID NO: 35 Amino acids 24-33 of HPV E6

TABLE-US-00053 HDIILECVY

[0437] SEQ ID NO: 36 Amino acids 26-35 of HPV E6

TABLE-US-00054 IILECVYCK

[0438] SEQ ID NO: 37 Amino acids 35-44 of HPV E6

TABLE-US-00055 QQLLRREVY

[0439] SEQ ID NO: 38 Amino acids 42-51 of HPV E6

TABLE-US-00056 VYDFARRDL

[0440] SEQ ID NO: 39 Amino acids 43-52 of HPV E6

TABLE-US-00057 YDFARRDLC

[0441] SEQ ID NO: 40 Amino acids 44-53 of HPV E6

TABLE-US-00058 DFARRDLCI

[0442] SEQ ID NO: 41 Amino acids 45-54 of HPV E6

TABLE-US-00059 FARRDLCIV

[0443] SEQ ID NO: 42 Amino acids 60-69 of HPV E6

TABLE-US-00060 YAVRDKCLK

[0444] SEQ ID NO: 43 Amino acids 62-71 of HPV E6

TABLE-US-00061 VRDKCLKFY

[0445] SEQ ID NO: 44 Amino acids 81-90 of HPV E6

TABLE-US-00062 YSLYGTTLE

[0446] SEQ ID NO: 45 Amino acids 125-134 of HPV E6

TABLE-US-00063 FHNIRGRWT

[0447] SEQ ID NO: 46 Amino acids 131-140 of HPV E6

TABLE-US-00064 RWTGRCMSC

[0448] SEQ ID NO: 47 Amino acids 143-152 of HPV E6

TABLE-US-00065 SRTRRETQL

[0449] SEQ ID NO: 48 Amino acids 16-31 of HPV E6

TABLE-US-00066 CTELQTTIHDIILEC

[0450] SEQ ID NO: 49 Amino acids 17-32 of HPV E6

TABLE-US-00067 TELQTTIHDIILECV

[0451] SEQ ID NO: 50 Amino acids 18-33 of HPV E6

TABLE-US-00068 ELQTTIHDIILECVY

[0452] SEQ ID NO: 51 Amino acids 19-34 of HPV E6

TABLE-US-00069 LQTTIHDIILECVYC

[0453] SEQ ID NO: 52 Amino acids 20-35 of HPV E6

TABLE-US-00070 QTTIHDIILECVYCK

[0454] SEQ ID NO: 53 Amino acids 34-49 of HPV E6

TABLE-US-00071 KQQLLRREVYDFARR

[0455] SEQ ID NO: 54 Amino acids 49-64 of HPV E6

TABLE-US-00072 DLCIVYRDGNPYAVR

[0456] SEQ ID NO: 55 Amino acids 50-65 of HPV E6

TABLE-US-00073 LCIVYRDGNPYAVRD

[0457] SEQ ID NO: 56 Amino acids 51-66 of HPV E6

TABLE-US-00074 CIVYRDGNPYAVRDK

[0458] SEQ ID NO: 57 Amino acids 78-93 of HPV E6

TABLE-US-00075 HYCYSLYGTTLEQQY

[0459] SEQ ID NO: 58 Amino acids 95-110 of HPV E6

TABLE-US-00076 PLCDLLIRCINRQKP

[0460] SEQ ID NO: 59 Amino acids 96-111 of HPV E6

TABLE-US-00077 LCDLLIRCINRQKPL

[0461] SEQ ID NO: 60 Amino acids 97-112 of HPV E6

TABLE-US-00078 CDLLIRCINRQKPLC

[0462] SEQ ID NO: 61 Amino acids 98-113 of HPV E6

TABLE-US-00079 DLLIRCINRQKPLCP

[0463] SEQ ID NO: 62 Amino acids 99-114 of HPV E6

TABLE-US-00080 LLIRCINRQKPLCPE

[0464] SEQ ID NO: 63 Amino acids 121-136 of HPV E6

TABLE-US-00081 KKQRFHNIRGRWTGR

[0465] SEQ ID NO: 64 Amino acids 7-16 of HPV E7

TABLE-US-00082 TLHEYMLDL

[0466] SEQ ID NO: 65 Amino acids 22-31 of HPV E7

TABLE-US-00083 LYGYGQLND

[0467] SEQ ID NO: 66 Amino acids 38-47 of HPV E7

TABLE-US-00084 IDGPAGQAE

[0468] SEQ ID NO: 67 Amino acids 48-57 of HPV E7

TABLE-US-00085 DRAHYNIVT

[0469] SEQ ID NO: 68 Amino acids 55-64 of HPV E7

TABLE-US-00086 VTFCCKCDS

[0470] SEQ ID NO: 69 Amino acids 66-75 of HPV E7

TABLE-US-00087 RLCVQSTHV

[0471] SEQ ID NO: 70 Amino acids 72-81 of HPV E7

TABLE-US-00088 THVDIRTLE

[0472] SEQ ID NO: 71 Amino acids 73-82 of HPV E7

TABLE-US-00089 HVDIRTLED

[0473] SEQ ID NO: 72 Amino acids 85-94 of HPV E7

TABLE-US-00090 GTLGIVCPI

[0474] SEQ ID NO: 73 Amino acids 4-19 of HPV E7

TABLE-US-00091 DTPTLHEYMLDLQPE

[0475] SEQ ID NO: 74 Amino acids 5-20 of HPV E7

TABLE-US-00092 TPTLHEYMLDLQPET

[0476] SEQ ID NO: 75 Amino acids 6-21 of HPV E7

TABLE-US-00093 PTLHEYMLDLQPETT

[0477] SEQ ID NO: 76 Amino acids 7-22 of HPV E7

TABLE-US-00094 TLHEYMLDLQPETTD

[0478] SEQ ID NO: 77 Amino acids 8-23 of HPV E7

TABLE-US-00095 LHEYMLDLQPETTDL

[0479] SEQ ID NO: 78 Amino acids 9-24 of HPV E7

TABLE-US-00096 HEYMLDLQPETTDLY

[0480] SEQ ID NO: 79 Amino acids 10-25 of HPV E7

TABLE-US-00097 EYMLDLQPETTDLYG

[0481] SEQ ID NO: 80 Amino acids 70-85 of HPV E7

TABLE-US-00098 QSTHVDIRTLEDLLM

[0482] SEQ ID NO: 81 Amino acids 71-86 of HPV E7

TABLE-US-00099 STHVDIRTLEDLLMG

[0483] SEQ ID NO: 82 Amino acids 72-87 of HPV E7

TABLE-US-00100 THVDIRTLEDLLMGT

[0484] SEQ ID NO: 83 Amino acids 73-88 of HPV E7

TABLE-US-00101 HVDIRTLEDLLMGTL

[0485] SEQ ID NO: 84 Amino acids 74-89 of HPV E7

TABLE-US-00102 VDIRTLEDLLMGTLG

[0486] SEQ ID NO: 85 Amino acids 16-25 of HPV E6

TABLE-US-00103 CTELQTTIH

[0487] SEQ ID NO: 86 Amino acids 42-51 of HPV E6

TABLE-US-00104 VYDFARRDL

[0488] SEQ ID NO: 87 Amino acids 45-54 of HPV E6

TABLE-US-00105 FARRDLCIV

[0489] SEQ ID NO: 88 Amino acids 52-61 of HPV E6

TABLE-US-00106 IVYRDGNPY

[0490] SEQ ID NO: 89 Amino acids 54-63 of HPV E6

TABLE-US-00107 YRDGNPYAV

[0491] SEQ ID NO: 90 Amino acids 68-77 of HPV E6

TABLE-US-00108 KFYSKISEY

[0492] SEQ ID NO: 91 Amino acids 73-82 of HPV E6

TABLE-US-00109 ISEYRHYCY

[0493] SEQ ID NO: 92 Amino acids 75-84 of HPV E6

TABLE-US-00110 EYRHYCYSL

[0494] SEQ ID NO: 93 Amino acids 80-89 of HPV E6

TABLE-US-00111 CYSLYGTTL

[0495] SEQ ID NO: 94 Amino acids 84-93 of HPV E6

TABLE-US-00112 YGTTLEQQY

[0496] SEQ ID NO: 95 Amino acids 88-97 of HPV E6

TABLE-US-00113 LEQQYNKPL

[0497] SEQ ID NO: 96 Amino acids 91-100 of HPV E6

TABLE-US-00114 QYNKPLCDL

[0498] SEQ ID NO: 97 Amino acids 11-20 of HPV E6

TABLE-US-00115 RHLDKKQRF

[0499] SEQ ID NO: 98 Amino acids 12-21 of HPV E6

TABLE-US-00116 RFHNIRGRW

[0500] SEQ ID NO: 99 Amino acids 18-33 of HPV E6

TABLE-US-00117 ELQTTIHDIILECVY

[0501] SEQ ID NO: 100 Amino acids 49-64 of HPV E6

TABLE-US-00118 DLCIVYRDGNPYAVR

[0502] SEQ ID NO: 101 Amino acids 50-65 of HPV E6

TABLE-US-00119 LCIVYRDGNPYAVRD

[0503] SEQ ID NO: 102 Amino acids 51-66 of HPV E6

TABLE-US-00120 CIVYRDGNPYAVRDK

[0504] SEQ ID NO: 103 Amino acids 74-89 of HPV E6

TABLE-US-00121 SEYRHYCYSLYGTTL

[0505] SEQ ID NO: 104 Amino acids 75-90 of HPV E6

TABLE-US-00122 EYRHYCYSLYGTTLE

[0506] SEQ ID NO: 105 Amino acids 76-91 of HPV E6

TABLE-US-00123 YRHYCYSLYGTTLEQ

[0507] SEQ ID NO: 106 Amino acids 77-92 of HPV E6

TABLE-US-00124 RHYCYSLYGTTLEQQ

[0508] SEQ ID NO: 107 Amino acids 78-93 of HPV E6

TABLE-US-00125 HYCYSLYGTTLEQQY

[0509] SEQ ID NO: 108 Amino acids 96-111 of HPV E6

TABLE-US-00126 LCDLLIRCINRQKPL

[0510] SEQ ID NO: 109 Amino acids 97-112 of HPV E6

TABLE-US-00127 CDLLIRCINRQKPLC

[0511] SEQ ID NO: 110 Amino acids 98-113 of HPV E6

TABLE-US-00128 DLLIRCINRQKPLCP

[0512] SEQ ID NO: 111 Amino acids 99-114 of HPV E6

TABLE-US-00129 LLIRCINRQKPLCPE

[0513] SEQ ID NO: 112 Amino acids 121-136 of HPV E6

TABLE-US-00130 KKQRFHNIRGRWTGR

[0514] SEQ ID NO: 113 Amino acids 3-12 of HPV E7

TABLE-US-00131 GDTPTLHEY

[0515] SEQ ID NO: 114 Amino acids 7-16 of HPV E7

TABLE-US-00132 TLHEYMLDL

[0516] SEQ ID NO: 115 Amino acids 9-18 of HPV E6

TABLE-US-00133 HEYMLDLQP

[0517] SEQ ID NO: 116 Amino acids 15-24 of HPV E6

TABLE-US-00134 LQPETTDLY

[0518] SEQ ID NO: 117 Amino acids 19-28 of HPV E6

TABLE-US-00135 TTDLYGYGQ

[0519] SEQ ID NO: 118 Amino acids 20-29 of HPV E6

TABLE-US-00136 TDLYGYGQL

[0520] SEQ ID NO: 119 Amino acids 44-53 of HPV E6

TABLE-US-00137 QAEPDRAHY

[0521] SEQ ID NO: 120 Amino acids 49-58 of HPV E6

TABLE-US-00138 RAHYNIVTF

[0522] SEQ ID NO: 121 Amino acids 71-80 of HPV E6

TABLE-US-00139 STHVDIRTL

[0523] SEQ ID NO: 122 Amino acids 79-88 of HPV E6

TABLE-US-00140 LEDLLMGTL

[0524] SEQ ID NO: 123 Amino acids ‘03-18 of HPV E6

TABLE-US-00141 GDTPTLHEYMLDLQP

[0525] SEQ ID NO: 124 Amino acids 4-19 of HPV E6

TABLE-US-00142 DTPTLHEYMLDLQPE

[0526] SEQ ID NO: 125 Amino acids 5-20 of HPV E6

TABLE-US-00143 TPTLHEYMLDLQPET

[0527] SEQ ID NO: 126 Amino acids 6-21 of HPV E6

TABLE-US-00144 PTLHEYMLDLQPETT

[0528] SEQ ID NO: 127 Amino acids 7-22 of HPV E6

TABLE-US-00145 TLHEYMLDLQPETTD

[0529] SEQ ID NO: 128 Amino acids 8-23 of HPV E6

TABLE-US-00146 LHEYMLDLQPETTDL

[0530] SEQ ID NO: 129 Amino acids 70-85 of HPV E6

TABLE-US-00147 QSTHVDIRTLEDLLM

[0531] SEQ ID NO: 130 Amino acids 71-86 of HPV E6

TABLE-US-00148 STHVDIRTLEDLLMG

[0532] SEQ ID NO: 131 Amino acids 72-87 of HPV E6

TABLE-US-00149 THVDIRTLEDLLMGT

[0533] SEQ ID NO: 132 Amino acids 73-88 of HPV E6

TABLE-US-00150 HVDIRTLEDLLMGTL

[0534] SEQ ID NO: 133 Amino acids 74-89 of HPV E6

TABLE-US-00151 VDIRTLEDLLMGTLG

[0535] SEQ ID NO: 134 T1D320

TABLE-US-00152 HNEGDDQQGSRYSLIPQIQKVCEVVDG

[0536] SEQ ID NO: 135 T1D814

TABLE-US-00153 SEQERMKSSLLKEHMLRKQAELESAQC

[0537] SEQ ID NO: 136 T1D182

TABLE-US-00154 ELRENLLTYLPDSVTQLRRLEELDLGN

[0538] SEQ ID NO: 137 T1D689

TABLE-US-00155 QEVFSSYKFNHLVRRLVLQREKHFHYL

[0539] SEQ ID NO: 138 T1D339

TABLE-US-00156 IEKNADLCYLSTVHWSLILDAVSNNYI

[0540] SEQ ID NO: 139 T1D428

TABLE-US-00157 LATAGEPYHDIRFKLMAVVPDRRIKYE

[0541] SEQ ID NO: 140 T1D572

TABLE-US-00158 MLRLPTVFRQIRPVSRVLAPHLTR

[0542] SEQ ID NO: 141 T1D359

TABLE-US-00159 IQIMENPFVQSMLWNPDLMRQLIMANP

[0543] SEQ ID NO: 142 T1D488

TABLE-US-00160 LLRFLFLGLSALALPSRAQLQLHLPAN

[0544] SEQ ID NO: 143 T1D554

TABLE-US-00161 MGKNPVRPPRAFPPVPSIDDIPLSR

[0545] SEQ ID NO: 144 T1D272

TABLE-US-00162 GHVDFTIEVERALTVLDGAVLVLCAVG

[0546] SEQ ID NO: 145 T1D210

TABLE-US-00163 EYKLMYGMLFSIRLFVSKMSPLDMKDG

[0547] SEQ ID NO: 146 T1D849

TABLE-US-00164 SQGAGVESLDFRLYLRYEFLMLGIQPV

[0548] SEQ ID NO: 147 T1D4

TABLE-US-00165 AAKTLVLCVSDIILLSANISETSSNKT

[0549] SEQ ID NO: 148 T1D77

TABLE-US-00166 CKDLQLYLSNLANHIDRETGIGDVPLV

[0550] SEQ ID NO: 149 T1D717

TABLE-US-00167 QSKHTEARELMYSAALLFFSHGQQNSA

[0551] SEQ ID NO: 150 T1D586

TABLE-US-00168 MTLAESYAQYVYNLCNSLSIKVEES

[0552] SEQ ID NO: 151 T11D323

TABLE-US-00169 KKVSKTRHTRETVFRRAKRRWAPIPCS

[0553] SEQ ID NO: 152 T11D506

TABLE-US-00170 NLKHENILQFLTAQERKTELGKQYWLI

[0554] SEQ ID NO: 153 T11D12

TABLE-US-00171 VQVFGLYFGEEFHETFDCPlK

[0555] SEQ ID NO: 154 T11D315

TABLE-US-00172 GQAAASQAGGARGYARGAQLWPPGSDP

[0556] SEQ ID NO: 155 T11D302

TABLE-US-00173 SIMQTWFTLFTPTDATSIVATTVMSNS

[0557] SEQ ID NO: 156 T11D700

TABLE-US-00174 QENVRFVLVRSEALLPNAGPRSAEARV

[0558] SEQ ID NO: 157 T11D535

TABLE-US-00175 DAFPNLKDFISRFQVMPPSSFLFDAPC

[0559] SEQ ID NO: 158 T11D358

TABLE-US-00176 PSSLQVKPETPASEAVAVAAAAAPTTT

[0560] SEQ ID NO: 159 T11D670

TABLE-US-00177 RFFHLADLFLSSSQLPAYLVAAFAKRL

[0561] SEQ ID NO: 160 T11D294

TABLE-US-00178 LVFLWLHSLRRLFGCLYVSVFSNVMIH

[0562] SEQ ID NO: 161 T11D336

TABLE-US-00179 WENAIAALFRRHIAVSWLIRATLSESE

[0563] SEQ ID NO: 162 T11D499

TABLE-US-00180 RYIFVKSAGSRIEEGVLQFLVLLVAGR

[0564] SEQ ID NO: 163 T11D425

TABLE-US-00181 QEIEWLPFRCIKCLKLSFSTAELLCMH

[0565] SEQ ID NO: 164 T11D491

TABLE-US-00182 LVARCPPCLRLLRQARDFQAARYDRHD

[0566] SEQ ID NO: 165 T11D314

TABLE-US-00183 ALGTPEDLDSYIDLSLESLNQMILELD

[0567] SEQ ID NO: 166 T11D430

TABLE-US-00184 SQDMLSIMEKLEFLDFSYDLNLCGLTE

[0568] SEQ ID NO: 167 T11D582

TABLE-US-00185 RREQYIPNEEFLHFDLLEDSKYRKIYS

[0569] SEQ ID NO: 168 T1D223

TABLE-US-00186 GPSYRSNSVSSLDLEGE

[0570] SEQ ID NO: 169 T1D164

TABLE-US-00187 LKEEKENLQGLVTHQTYIIQELEKQLN

[0571] SEQ ID NO: 170 T1D56

TABLE-US-00188 QRQKTAGKIFRAEVSTGQDAPRQAQAR

[0572] SEQ ID NO: 171 T1D36

TABLE-US-00189 ILRPSTVNELESGEISYDEVGERIKDF

[0573] SEQ ID NO: 172 T1D129

TABLE-US-00190 DTCINEDVESLRKMVQDLLAKLQEAKR

[0574] SEQ ID NO: 173 T1D274

TABLE-US-00191 LREEGTKYKSFVQRARLVFREEGYLAF

[0575] SEQ ID NO: 174 T1D62

TABLE-US-00192 HEGARPMRAIFLANGNVFTTGFSRMSE

[0576] SEQ ID NO: 175 T1D5

TABLE-US-00193 IPDAAFIQAARRKCELARAQDDYISLD

[0577] SEQ ID NO: 176 T1D144

TABLE-US-00194 MEIAKMRAGRRLWVHLIEKMFQPKNSK

[0578] SEQ ID NO: 177 T1D441

TABLE-US-00195 MELVSFRDVAIEFSPEEW

[0579] SEQ ID NO: 178 T1D368

TABLE-US-00196 TCVHEIPFHFDLMELLPQCQQLQMFFL

[0580] SEQ ID NO: 179 T1D234

TABLE-US-00197 VRRVFITDDFHDMIPKYLNFVKGVVDS

[0581] SEQ ID NO: 180 T1D162

TABLE-US-00198 NFHAHREKAPSLFCSRILNKAVYLFYG

[0582] SEQ ID NO: 181 T1D39

TABLE-US-00199 DANKEGMFLFRAAHKLRQFLKMNSTGD

[0583] SEQ ID NO: 182 T1D272

TABLE-US-00200 LLSLGWSVDVGRHSGWTGHVSTSWSIN

[0584] SEQ ID NO: 183 T1D328

TABLE-US-00201 NTIFSLRKDLRQGKARRLRCMEEKEMF

[0585] SEQ ID NO: 184 T1D188

TABLE-US-00202 NHFNDFEGDPAMTQFLEEFEKNLEDTK

[0586] SEQ ID NO: 185 C-pepM1

TABLE-US-00203 VILPQAPSGPSYATYLQPAQAQMLTPP

[0587] SEQ ID NO: 186 C-pepM6

TABLE-US-00204 DTLSAMSNPRAMQVLLQIQQGLQTLAT

[0588] SEQ ID NO: 187 C-pepM8

TABLE-US-00205 EVIQTSKYYMRDVIAIESAWLLELAPH

[0589] SEQ ID NO: 188 C-pepM29

TABLE-US-00206 GSLFGSSRVQYVVNPAVKIVFLNIDPS

[0590] SEQ ID NO: 189 C-pepM31

TABLE-US-00207 AEYGDYQPEVHGVPYFRLEHYLPARVM

[0591] SEQ ID NO: 190 C-pepM43

TABLE-US-00208 KSVVIHCWKYLSVQSQLFRGSSLLFRRV

[0592] SEQ ID NO: 191 C-pepM89

TABLE-US-00209 TLAFLVLSTPAMFNRALKPFLKSCHFQ

[0593] SEQ ID NO: 192 C-pep149

TABLE-US-00210 FVSPMAHYVPGIMAIESVVARFQFIVP

[0594] SEQ ID NO: 193 C-pepM171

TABLE-US-00211 LWVYLRPVPRPATIYLQILRLKPL TGE

[0595] SEQ ID NO: 194 C-pepM173

TABLE-US-00212 KlYEFDYHLYGQNITMIMTSVSGHLLA

[0596] SEQ ID NO: 195 VB 4101 Amino acids 81-88

TABLE-US-00213 RGPESRLL

[0597] SEQ ID NO: 196 VB4102 Amino Acids 81-126

TABLE-US-00214 RGPESRLLEFYLAMPFATPMEAELARRSLAQDAPPLPVPGVLLKEF

[0598] SEQ ID NO: 197 NY-ESO_Pep-1

TABLE-US-00215 MQAEGRGTGGSTGDA

[0599] SEQ ID NO: 198 NY-ESO_Pep-2

TABLE-US-00216 TGGSTGDADGPGGPG

[0600] SEQ ID NO: 199 NY-ESO_Pep-3

TABLE-US-00217 ADGPGGPGIPDGPGG

[0601] SEQ ID NO: 200 NY-ESO_Pep-4

TABLE-US-00218 GIPDGPGGNAGGPGE

[0602] SEQ ID NO: 201 NY-ESO_Pep-5

TABLE-US-00219 GNAGGPGEAGATGGR

[0603] SEQ ID NO: 202 NY-ESO_Pep-6

TABLE-US-00220 EAGATGGRGPRGAGA

[0604] SEQ ID NO: 203 NY-ESO_Pep-7

TABLE-US-00221 RGPRGAGAARASGPG

[0605] SEQ ID NO: 204 NY-ESO_Pep-8

TABLE-US-00222 AARASGPGGGAPRGP

[0606] SEQ ID NO: 205 NY-ESO_Pep-9

TABLE-US-00223 GGGAPRGPHGGAASG

[0607] SEQ ID NO: 206 NY-ESO_Pep-10

TABLE-US-00224 PHGGAASGLNGCCRC

[0608] SEQ ID NO: 207 NY-ESO_Pep-11

TABLE-US-00225 GLNGCCRCGARGPES

[0609] SEQ ID NO: 208 NY-ESO_Pep-12

TABLE-US-00226 CGARGPESRLLEFYL

[0610] SEQ ID NO: 209 NY-ESO_Pep-13

TABLE-US-00227 RGPESRLLEFYLAMP

[0611] SEQ ID NO: 210 NY-ESO_Pep-14

TABLE-US-00228 SRLLEFYLAMPFATP

[0612] SEQ ID NO: 211 NY-ESO_Pep-15

TABLE-US-00229 LAMPFATPMEAELAR

[0613] SEQ ID NO: 212 NY-ESO_Pep-16

TABLE-US-00230 PMEAELARRSLAQDA

[0614] SEQ ID NO: 213 NY-ESO_Pep-17

TABLE-US-00231 RRSLAQDAPPLPVPG

[0615] SEQ ID NO: 214 NY-ESO_Pep-18

TABLE-US-00232 DAPPLPVPGVLLKEF

[0616] SEQ ID NO: 215 NY-ESO_Pep-19

TABLE-US-00233 APPLPVPGVLLKEFT

[0617] SEQ ID NO: 216 NY-ESO_Pep-20

TABLE-US-00234 GVLLKEFTVSGNILT

[0618] SEQ ID NO: 217 NY-ESO_Pep-21

TABLE-US-00235 TVSGNILTIRLTAAD

[0619] SEQ ID NO: 218 NY-ESO_Pep-22

TABLE-US-00236 TIRLTAADHRQLQLS

[0620] SEQ ID NO: 219 NY-ESO_Pep-23

TABLE-US-00237 DHRQLQLSISSCLQQ

[0621] SEQ ID NO: 220 NY-ESO_Pep-24

TABLE-US-00238 SISSCLQQLSLLMWI

[0622] SEQ ID NO: 221 NY-ESO_Pep-25

TABLE-US-00239 QLSLLMWITQCFLPV

[0623] SEQ ID NO: 222 NY-ESO_Pep-26

TABLE-US-00240 ITQCFLPVFLAQPPS

[0624] SEQ ID NO: 223 NY-ESO_Pep-27

TABLE-US-00241 VFLAQPPSGQRR

[0625] SEQ ID NO: 224 NY-ESO_Pep-28

TABLE-US-00242 RGPESRLL

[0626] SEQ ID NO: 225 B-pepM2

TABLE-US-00243 REGVELCPGNKYEMRRHGTTHSLVIHD

[0627] SEQ ID NO: 226 B-pepM7

TABLE-US-00244 SSPDEVALVEGVQSLGFTYLRLKDNYM

[0628] SEQ ID NO: 227 B-pepM36

TABLE-US-00245 KNILAVSFAPLVQLSKNDNGTPDSVGL

[0629] SEQ ID NO: 228 B-pepM78

TABLE-US-00246 PAADRWEPRAPMRAPRVLHAMLGAAGR

[0630] SEQ ID NO: 229 B-pepM79

TABLE-US-00247 VLSADLVHYLRLSLEYLRAWHSEDVSL

[0631] SEQ ID NO: 230 B-pepM82

TABLE-US-00248 LEAVLRYLETHPRLPKPDPVKSSSSVL

[0632] SEQ ID NO: 231 B-pepM83

TABLE-US-00249 LKDFFFSRGRFSSALKRTLILSSGFRL

[0633] SEQ ID NO: 232 B-pepM84

TABLE-US-00250 AGKDHKIKQWDADTFEHIQTLEGHHQE

[0634] SEQ ID NO: 233 B-pepM85

TABLE-US-00251 PATIEMAIETLQKFDGLSTHRSSLLNS

[0635] SEQ ID NO: 234 B-pepM86

TABLE-US-00252 YLPGGGAGHLDQNVFLRVRALCYVISG

[0636] SEQ ID NO: 235 VB4119 Amino acids 180-188

TABLE-US-00253 SVYDFFVWL

[0637] SEQ ID NO: 236 B-pepM108

TABLE-US-00254 YISDHMKVHSPSPCL

[0638] SEQ ID NO: 237 B-pepM115-M122

TABLE-US-00255 EGWQTCWGRSRKHWGSTWNGSARLSPGSTLWVMRICLRSLGIARTWLSCR STSRKCSPAFPASS

[0639] SEQ ID NO: 238 B-pepM141-M142

TABLE-US-00256 LFRLLPSGPKVNDGHRSRRWHSLDLYPHLCSHLDLPKQSQEVIRL

[0640] SEQ ID NO: 239 B-pepM108

TABLE-US-00257 YISDHMKVHSPSPCL

[0641] SEQ ID NO: 240 B-pepM115

TABLE-US-00258 EGWQTCWGRSRKHWG

[0642] SEQ ID NO: 241 B-pepM116

TABLE-US-00259 GRSRKHWGSTWNGSA

[0643] SEQ ID NO: 242 B-pepM117

TABLE-US-00260 GSTWNGSARLSPGST

[0644] SEQ ID NO: 243 B-pepM118

TABLE-US-00261 ARLSPGSTLWVMRIC

[0645] SEQ ID NO: 244 B-pepM119

TABLE-US-00262 TLWVMRICLRSLGIA

[0646] SEQ ID NO: 245 B-pepM120

TABLE-US-00263 CLRSLGIARTWLSCR

[0647] SEQ ID NO: 246 B-pepM121

TABLE-US-00264 ARTWLSCRSTSRKCS

[0648] SEQ ID NO: 247 B-pepM122

TABLE-US-00265 RSTSRKCSPAFPASS

[0649] SEQ ID NO: 248 B-pepM141

TABLE-US-00266 LFRLLPSGPKVNDGHRSRRWHSLDLYP

[0650] SEQ ID NO: 249 B-pepM142

TABLE-US-00267 SRRWHSLDLYPHLCSHLDLPKQSQEVIRL

Embodiments A

[0651] 1. An individualized therapeutic anticancer vaccine comprising an immunologically effective amount of: [0652] (i) a polynucleotide comprising a nucleotide sequence encoding a targeting unit, a dimerization unit and an antigenic unit, wherein said antigenic unit comprises at least one patient-present shared antigen sequence or one or more parts thereof, and optionally one or more patient-specific antigen sequences or one or more parts thereof; or [0653] (ii) a polypeptide encoded by the polynucleotide as defined in (i), or [0654] (iii) a dimeric protein consisting of two polypeptides encoded by the polynucleotide as defined in (i); anda pharmaceutically acceptable carrier.

[0655] 2. The vaccine according to embodiment A1, wherein said at least one patient-present shared antigen sequence is a shared antigen selected from the group consisting of overexpressed cellular proteins, aberrantly expressed cellular proteins, cancer testis antigens, viral antigens, differentiation antigens, mutated oncogenes and mutated tumor suppressor genes, oncofetal antigens, shared fusion antigens, shared intron retention antigens, dark matter antigens and shared antigens caused by spliceosome mutations or frameshift mutations

[0656] 3. The vaccine according to any of embodiments A1 or A2, wherein said at least one patient-present shared antigen sequence is a shared antigen that is a human cellular protein, preferably an overexpressed or aberrantly expressed human cellular protein or a differentiation antigen.

[0657] 4. The vaccine according to any of embodiments A1 to A3, wherein said at least one patient-present shared antigen sequence is known to be immunogenic or wherein one or more parts thereof are known to be immunogenic or are predicted to bind to the patient’s HLA class | or HLA class II alleles, preferably to the patient’s HLA class I alleles.

[0658] 5. The vaccine according to any of embodiments A1 to A4, wherein said at least one patient-present shared antigen sequence has a length suitable for presentation by the patient’s HLA alleles, preferably a length of from 7 to 30 amino acids.

[0659] 6. The vaccine according to any of embodiments A1 to A5, comprising more than one patient-present shared antigen sequence or one or more parts thereof.

[0660] 7. The vaccine according to embodiment A6, comprising sequences of several patient-present shared antigens or one or more parts thereof, preferably several parts of sequences of several patient-present shared antigens, more preferably several epitopes of several patient-present shared antigen, which epitopes are known to be immunogenic or are predicted to bind to the particular patient’s HLA class | and HLA class II alleles.

[0661] 8. The vaccine according to any of embodiments A1 to A7, wherein the antigenic unit comprises one or more patient-present shared antigen sequences in full length, preferably 1 to 10 patient-present shared antigen sequences in full length.

[0662] 9. The vaccine according to any of embodiments A1 to A7, wherein the antigenic unit comprises 1 to 30 parts of patient-present shared antigen sequences in the form of long peptide sequences, preferably peptide sequences of from about 28 to 100 amino acids.

[0663] 10. The vaccine according to embodiment A9, wherein the long peptide sequences include multiple epitopes that are predicted to bind to the patient’s HLA class | or HLA class II alleles.

[0664] 11. The vaccine according to any of embodiments A1 to A7, wherein the antigenic unit comprises 1 to 50 parts of patient-present shared antigen sequences in the form of short peptide sequences/epitopes.

[0665] 12. The vaccine according to embodiment A11, wherein the short peptide sequences/epitopes that are predicted to bind to the patient’s HLA class | or HLA class II alleles.

[0666] 13. The vaccine according to any of embodiments A11 to A12, wherein the short peptide sequences/epitopes have a length of from 7 to 30 amino acids, e.g. 7 to 10 or 13 to 30.

[0667] 14. The vaccine according to any of the preceding embodiments A1 to A13, comprising one or more patient-specific antigen sequences or one or more parts thereof.

[0668] 15. The vaccine according to embodiment A14, comprising several patient-specific antigen sequences or one or more parts thereof.

[0669] 16. The vaccine according to embodiment A15, comprising one or more parts of said patient-specific antigen sequences, preferably one or more patient-specific epitope.

[0670] 17. The vaccine according to embodiment A16, comprising one or more patient-specific epitopes having a length of from 7 to 30 amino acids, e.g. 7 to 10 or 13 to 30.

[0671] 18. The vaccine according to any of embodiments A14 to A17, wherein antigenic unit comprises at least 10 patient-specific epitopes, preferably at least 15 patient-specific epitopes, such as at least 20 patient-specific epitopes.

[0672] 19. The vaccine according to any of the preceding embodiments A1 to A18, wherein the antigenic unit comprises from 21 to 2000 amino acids, preferably from about 30 amino acids to about a 1500 amino acids, more preferably from about 50 to about 1000 amino acids, such as from about 100 to about 500 amino acids or from about 100 to about 400 amino acids or from about 100 to about 300 amino acids.

[0673] 20. The vaccine according to any of the preceding embodiments A1 to A19, wherein the antigenic unit comprises one or more linkers, preferably one or more non-immunogenic and/or flexible linkers.

[0674] 21. The vaccine according to embodiment A20, wherein the length of the one or more linkers is from 4 to 20 amino acids.

[0675] 22. The vaccine according to any of embodiments A20 to A21, wherein the one or more linkers separate the antigen sequences from each other.

[0676] 23. The vaccine according to any of the preceding embodiments A1 to A22, wherein the dimerization unit comprises a hinge region and optionally another domain that facilitates dimerization, optionally connected through a linker.

[0677] 24. The vaccine according to embodiment A23, wherein the hinge region is Ig-derived.

[0678] 25. The vaccine according to any one of embodiments A23 and A24, wherein the hinge region has the ability to form one or more covalent bonds, preferably a covalent bond in the form of a disulfide bridge.

[0679] 26. The vaccine according to any one of embodiments A23 to A25, wherein the another domain that facilitates dimerization is an immunoglobulin domain, preferably a carboxyterminal C domain, or a sequence that is substantially identical to said C domain or a variant thereof.

[0680] 27. The vaccine according to embodiment A26, wherein the carboxyterminal C domain is derived from IgG.

[0681] 28. The vaccine according to any one of embodiments A26 and A37, wherein the immunoglobulin domain of the dimerization unit has the ability to homodimerize, preferably via noncovalent interactions and more preferably via noncovalent interactions that are hydrophobic interactions.

[0682] 29. The vaccine according to any one of embodiments A23 to A28, wherein said dimerization unit does not comprise a CH2 domain.

[0683] 30. The vaccine according to any one of embodiments A23 to A29, wherein the dimerization unit consist of hinge exons h1 and h4 connected through said third linker to a C.sub.H3 domain of human IgG3.

[0684] 31. The vaccine according to any one of embodiments A23 to A30, wherein the dimerization unit comprises an amino acid sequence having at least 80% sequence identity to the amino acid sequence 94-237 of SEQ ID NO.: 3.

[0685] 32. The vaccine according to any of the preceding embodiments A1 to A31, wherein the antigenic unit and the dimerization unit are connected through a linker, preferably a linker that comprises a restriction site.

[0686] 33. The vaccine according to any of the preceding embodiments A1 to A32, wherein the targeting unit has affinity for a chemokine receptor selected from CCR1, CCR3 and CCR5.

[0687] 34. The vaccine according to any of the preceding embodiments A1 to A33, wherein said targeting unit comprises antibody binding regions with specificity for CD14, CD40, or Toll- like receptor or ligands, such as soluble CD40 ligand, or chemokines, such as RANTES or MIP-1a or bacterial antigens, such as flagellin.

[0688] 35. The vaccine according to any of embodiments A1 to A33, wherein said targeting unit has an affinity for MHC class II proteins, preferably MHC class II proteins, selected from the group consisting of anti-HLA-DP, anti-HLA-DR and anti-pan HLA class II

[0689] 36. The vaccine according to any of embodiments A1 to A33, wherein said targeting unit has an amino acid sequence having at least 80% sequence identity to the amino acid sequence 24-93 of SEQ ID NO.: 1.

[0690] 37. The vaccine according to any of the preceding embodiments A1 to A36, wherein said polynucleotide further encodes a signal peptide.

[0691] 38. The vaccine according to any of the preceding embodiments A1 to A37, wherein said targeting unit, dimerization unit and antigenic unit in said peptide are in the N-terminal to C-terminal order of targeting unit, dimerization unit and antigenic unit.

[0692] 39. The vaccine according to any of the preceding embodiments A1 to A38, wherein said polynucleotide sequence is human codon optimized.

[0693] 40. The vaccine according to any of the preceding embodiments A1 to A39, wherein said polynucleotide sequence is a DNA nucleotide sequence or RNA nucleotide sequence.

[0694] 41. A polynucleotide as defined in any of the embodiments A1 to A40.

[0695] 42. A vector comprising the polynucleotide according to embodiment A41.

[0696] 43. A host cell comprising the polynucleotide as defined in any of the embodiments A1 to A40 or comprising the vector according to embodiment A42.

[0697] 44. The polynucleotide according to embodiment A41 formulated for administration to a patient to induce production of a dimeric protein in said patient.

[0698] 45. A polypeptide encoded by the polynucleotide sequence as defined in any of the embodiments A1 to A40.

[0699] 46. A dimeric protein consisting of two polypeptides according to embodiment A45.

[0700] 47. The dimeric protein according to embodiment A46, being a homodimeric protein.

[0701] 48. The polynucleotide according to embodiment A41 or the polypeptide according to embodiment A45 or the dimeric protein according to any of embodiments A46 to A47 for use as a medicament.

[0702] 49. A method for preparing an individualized therapeutic anticancer vaccine comprising an immunologically effective amount of [0703] (i) a dimeric protein consisting of two polypeptides encoded by a polynucleotide comprising a nucleotide sequence encoding a targeting unit, a dimerization unit and an antigenic unit, wherein said antigenic unit comprises at least one patient-present shared antigen sequence or one or more parts thereof, and optionally one or more patient-specific antigen sequences or one or more parts thereof; or [0704] (ii) a polypeptide encoded by a polynucleotide comprising a nucleotide sequence encoding a targeting unit, a dimerization unit and an antigenic unit, wherein said antigenic unit comprises at least one patient-present shared antigen sequence or one or more parts thereof, and optionally one or more patient-specific antigen sequences or one or more parts thereof, the method comprises: [0705] a) transfecting cells with the polynucleotide; [0706] b) culturing the cells; [0707] c) collecting and purifying the dimeric protein or the polypeptide expressed from the cells, and [0708] d) mixing the dimeric protein or polypeptide obtained from step c) with a pharmaceutically acceptable carrier.

[0709] 50. A method for preparing an individualized therapeutic anticancer vaccine comprising an immunologically effective amount of a polynucleotide comprising a nucleotide sequence encoding a targeting unit, a dimerization unit and an antigenic unit, wherein said antigenic unit comprises at least one patient-present shared antigen sequence or one or more parts thereof, and optionally one or more patient-specific antigen sequences or one or more parts thereof, the method comprises: [0710] a. preparing the polynucleotide; [0711] b. optionally cloning the polynucleotide into an expression vector and [0712] c. mixing the polynucleotide from step a) or the vector form step b) with a pharmaceutically acceptable carrier.

[0713] 51. The method according to embodiments A49 or A50, including the steps of identifying patient-present shared antigens and patient-specific antigens, identifying the patient’s HLA class | and HLA class II alleles, select patient-present shared antigen sequences and optionally patient-specific antigen sequences based on immunogenicity prior to preparing the polynucleotide.

[0714] 52. A method of treating cancer in a patient, the method comprising administering to the said patient the vaccine according to any of embodiments A1 to A40.

[0715] 53. The method according to embodiment A52, wherein the vaccine comprises a polynucleotide and is administered intradermally or intramuscularly.

[0716] 54. The method according to embodiment A53 wherein the polynucleotide is a DNA.

[0717] 55. The method according to embodiment A53 wherein the polynucleotide is an RNA.

[0718] 56. The method according to any of claims A52 to A55, wherein administration is carried out with a jet injector.

[0719] 57. The method according to any of claims A52 to A56, wherein administration is assisted by electroporation.

Embodiments B

[0720] 1. An individualized therapeutic anticancer vaccine comprising an immunologically effective amount of: [0721] (i) a polynucleotide comprising a nucleotide sequence encoding a targeting unit, a dimerization unit and an antigenic unit, wherein said antigenic unit comprises at least one patient-present shared antigen sequence or one or more parts thereof; or [0722] (ii) a polypeptide encoded by the polynucleotide as defined in (i); or [0723] (iii) a dimeric protein consisting of two polypeptides encoded by the polynucleotide as defined in (i); anda pharmaceutically acceptable carrier.

[0724] 2. The vaccine according to embodiment B1, wherein said antigenic unit further comprises one or more patient-specific antigen sequences or one or more parts thereof.

[0725] 3. The vaccine according to any of embodiments B1 or B2, wherein said at least one patient-present shared antigen sequence is a sequence of a shared antigen selected from the group consisting of overexpressed cellular proteins, aberrantly expressed cellular proteins, cancer testis antigens, viral antigens, differentiation antigens, mutated oncogenes, mutated tumor suppressor genes, oncofetal antigens, shared fusion antigens, shared intron retention antigens, dark matter antigens, shared antigens caused by spliceosome mutations and shared antigens caused by frameshift mutations.

[0726] 4. The vaccine according to any of embodiments B1 to B3, wherein said at least one patient-present shared antigen sequence is a sequence of a shared antigen that is a human cellular protein.

[0727] 5. The vaccine according to embodiment B4, wherein said human cellular protein is an overexpressed or aberrantly expressed human cellular protein or a differentiation antigen.

[0728] 6. The vaccine according to any of embodiments B1 to B5, wherein said at least one patient-present shared antigen sequence or one or more parts thereof are known to be immunogenic or are predicted to bind to the patient’s HLA class | and/or HLA class II alleles.

[0729] 7. The vaccine according to embodiment B6, wherein said at least one patient-present shared antigen sequence or one or more parts thereof are predicted to bind to the patient’s HLA class I alleles.

[0730] 8. The vaccine according to any of embodiments B1 to B7, wherein said at least one patient-present shared antigen sequence or the one or more parts thereof have a length suitable for presentation by the patient’s HLA alleles.

[0731] 9. The vaccine according to embodiment B8, wherein said at least one patient-present shared antigen sequence or the one or more parts thereof have a length of from 7 to 30 amino acids.

[0732] 10. The vaccine according to any of embodiments B1 to B9, wherein the antigenic unit comprises more than one patient-present shared antigen sequence or one or more parts thereof.

[0733] 11. The vaccine according to embodiment B10, wherein the antigenic unit comprises sequences of several patient-present shared antigens or one or more parts thereof.

[0734] 12. The vaccine according to embodiment B11, wherein the antigenic unit comprises several parts of sequences of several patient-present shared antigens.

[0735] 13. The vaccine according to embodiment B12, wherein the antigenic unit comprises several epitopes of several patient-present shared antigens, which epitopes are known to be immunogenic or are predicted to bind to the patient’s HLA class | and/or HLA class II alleles.

[0736] 14. The vaccine according to any of embodiments B1 to B13, wherein the antigenic unit comprises one or more patient-present shared antigen sequences in full length.

[0737] 15. The vaccine according to embodiment B14, wherein the antigenic unit comprises 1 to 10 patient-present shared antigen sequences in full length.

[0738] 16. The vaccine according to any of embodiments B1 to B15, wherein the antigenic unit comprises 1 to 30 parts of at least one patient-present shared antigen sequence.

[0739] 17. The vaccine according to embodiment B16, wherein said 1 to 30 parts have a length of from 28 to 100 amino acids.

[0740] 18. The vaccine according to embodiment B17, wherein the parts include multiple epitopes that are predicted to bind to the patient’s HLA class | and/or HLA class II alleles.

[0741] 19. The vaccine according to any of embodiments B1 to B18, wherein the antigenic unit comprises 1 to 50 patient-present shared antigen sequences in the form of epitopes.

[0742] 20. The vaccine according to embodiment B19, wherein the epitopes are predicted to bind to the patient’s HLA class | and/or HLA class II alleles.

[0743] 21. The vaccine according to any of embodiments B19 to B20, wherein the epitopes have a length of from 7 to 30 amino acids.

[0744] 22. The vaccine according to any of embodiments B2 to B21, wherein the antigenic unit comprises several patient-specific antigen sequences or one or more parts thereof.

[0745] 23. The vaccine according to embodiment B22, wherein the antigenic unit comprises one or more parts of said several patient-specific antigen sequences.

[0746] 24. The vaccine according to any of embodiments B2 to B23, wherein the antigenic unit comprises one or more patient-specific epitopes.

[0747] 25. The vaccine according to embodiment B24, wherein the one or more patient-specific epitopes have a length of from 7 to 30 amino acids.

[0748] 26. The vaccine according to any of embodiments B24 to B25, wherein antigenic unit comprises at least 5 patient-specific epitopes.

[0749] 27. The vaccine according to any of embodiments B24 to B25, wherein the antigenic unit comprises at least 10 patient-specific epitopes.

[0750] 28. The vaccine according to any of embodiments B24 to B25, wherein the antigenic unit comprises at least 15 patient-specific epitopes.

[0751] 29. The vaccine according to any of the preceding embodiments B1 to B28, wherein the antigenic unit comprises from 7 to 2000 amino acids.

[0752] 30. The vaccine according to embodiment B29, wherein the antigenic unit comprises from 30 to 1500 amino acids.

[0753] 31. The vaccine according to embodiment B29, wherein the antigenic unit comprises from 50 to 1000 amino acids.

[0754] 32. The vaccine according to any of the preceding embodiments B1 to B31, wherein the antigenic unit comprises one or more linkers.

[0755] 33. The vaccine according to embodiment B32, wherein the one or more linkers are non-immunogenic and/or flexible linkers.

[0756] 34. The vaccine according to any of embodiments B32 or B33, wherein the length of the one or more linkers is from 4 to 20 amino acids.

[0757] 35. The vaccine according to any of embodiments B32 to B34, wherein the one or more linkers separate the antigen sequences from each other.

[0758] 36. The vaccine according to any of the preceding embodiments B1 to B35, wherein the dimerization unit comprises a hinge region.

[0759] 37. The vaccine according to embodiment B36, wherein the hinge region has the ability to form one or more covalent bonds, preferably in the form of a disulfide bridge.

[0760] 38. The vaccine according to any of embodiments B36 or 3B7, wherein the hinge region is Ig derived.

[0761] 39. The vaccine according to any of embodiments B36 to B38, wherein the dimerization unit further comprises another domain that facilitates dimerization.

[0762] 40. The vaccine according to embodiment B39, wherein the other domain is an immunoglobulin domain, preferably an immunoglobulin constant domain.

[0763] 41. The vaccine according to any of embodiments B39 or B40, wherein the other domain is a carboxyterminal C domain derived from IgG, preferably derived from IgG3.

[0764] 42. The vaccine according to any of embodiments B36 to B41, wherein the dimerization unit further comprises a linker, preferably a linker that connects the hinge region and the other domain that facilitates dimerization.

[0765] 43. The vaccine according to any of embodiments B36 to B42, wherein the dimerization unit comprises hinge exons h1 and h4 connected through a linker to a CH3 domain of human IgG3.

[0766] 44. The vaccine according to any of embodiments B36 to B43, wherein the dimerization unit comprises an amino acid sequence having at least 80% sequence identity to the amino acid sequence 94-237 of SEQ ID NO: 3.

[0767] 45. The vaccine according to any of embodiments B36 to B44, wherein the dimerization unit consists of the amino acid sequence 94-237 of SEQ ID NO: 3.

[0768] 46. The vaccine according to any of the preceding embodiments B1 to B45, wherein the antigenic unit and the dimerization unit are connected through a linker, preferably a linker that comprises a restriction site.

[0769] 47. The vaccine according to any of the preceding embodiments B1 to B46, wherein the targeting unit targets antigen presenting cells.

[0770] 48. The vaccine according to embodiment B47, wherein the targeting unit is or comprises a moiety that interacts with surface molecules on the antigen presenting cells.

[0771] 49. The vaccine according to embodiment B48, wherein the surface molecule is selected from the group consisting of HLA, CD14, CD40, chemokine receptors and Toll-like receptors.

[0772] 50. The vaccine according to any of embodiments B47 to B49, wherein the targeting unit comprises or consists of soluble CD40 ligand, RANTES, MIP-1α, XCL1, XCL2, flagellin, anti-HLA-DP, anti-HLA-DR, anti-pan HLA class II or an antibody variable domain with specificity for anti-CD40, anti-TLR-2, anti-TLR-4 or anti-TLR-5.

[0773] 51. The vaccine according to embodiment B50, wherein the targeting unit comprises or consists of MIP-1α.

[0774] 52. The vaccine according to embodiment B51, wherein the targeting unit comprises an amino acid sequence having at least 80% sequence identity to the amino acid sequence 24-93 of SEQ ID NO: 1.

[0775] 53. The vaccine according to embodiment B52, wherein the targeting unit consists of an amino acid sequence having at least 80% sequence identity to the amino acid sequence 24-93 of SEQ ID NO: 1.

[0776] 54. The vaccine according to embodiment B53, wherein the targeting unit consist of the amino acid sequence 24-93 of SEQ ID NO: 1.

[0777] 55. The vaccine according to any of the preceding embodiments B1 to B54, wherein said vaccine comprises a polynucleotide, preferably an RNA or DNA.

[0778] 56. The vaccine according to embodiment B55, wherein said polynucleotide is human codon optimized.

[0779] 57. The vaccine according to any of embodiments B55 or B56, wherein the polynucleotide further comprises a nucleotide sequence encoding a signal peptide.

[0780] 58. The vaccine according to embodiment B57, wherein the signal peptide is selected from the list consisting of Ig VH signal peptide, human TPA signal peptide and human MIP1-α signal peptide.

[0781] 59. The vaccine according to any of embodiments B57 to B58, wherein the signal peptide comprises an amino acid sequence having at least 85% sequence identity to the amino acid sequence 1-23 of SEQ ID NO: 1.

[0782] 60. The vaccine according to embodiment B59, wherein the signal peptide consists of an amino acid sequence having at least 85% sequence identity to the amino acid sequence 1-23 of SEQ ID NO: 1.

[0783] 61. The vaccine according to embodiment B60, wherein the signal peptide consists of the amino acid sequence 1-23 of SEQ ID NO: 1.

[0784] 62. The vaccine according to any of the preceding embodiments B1 to B61, wherein said targeting unit, dimerization unit and antigenic unit in said polypeptide are in the N-terminal to C-terminal order of targeting unit, dimerization unit and antigenic unit or wherein said targeting unit, dimerization unit and antigenic unit in said polynucleotide are in the 5′ to 3′ order of targeting unit, dimerization unit and antigenic unit.

[0785] 63. The vaccine according to any of the preceding embodiments B1 to B62, wherein the pharmaceutically acceptable carrier is selected from the group consisting of saline, buffered saline, PBS, dextrose, water, glycerol, ethanol, sterile isotonic aqueous buffers, and combinations thereof.

[0786] 64. A method for preparing the individualized therapeutic anticancer vaccine of embodiment B1, said method comprises the steps of: [0787] a) identifying at least one patient-present shared antigen in the tumor tissue or body fluid of a patient [0788] b) determining the patient’s HLA class | and/or class II alleles [0789] c) predicting the immunogenicity of the identified at least one antigen or one or more parts thereof by their predicted binding to the patient’s HLA class | and/or II alleles [0790] d) selecting at least one antigen or one or more parts thereof based on their immunogenicity predicted in step c); [0791] e) preparing a polynucleotide sequence comprising an antigenic unit comprising a nucleotide sequence encoding the at least one antigen or one or more parts thereof selected in step d); [0792] f) cloning the polynucleotide sequence into an expression vector comprising nucleotide sequences encoding a targeting unit and a dimerization unit; and [0793] g) mixing the expression vector obtained in step f with a pharmaceutically acceptable carrier.

[0794] 65. The method according to embodiment B64 for preparing the individualized anticancer vaccine of embodiment B2, wherein said method further comprises: [0795] in step a) identifying one or more patient-specific antigens in the tumor tissue of the patient in step c) predicting the immunogenicity of the identified one or more patient-specific antigens or one or more parts thereof by their predicted binding to the patient’s HLA class | and/or II alleles in step d) selecting one or more patient-specific antigens or one or more parts thereof based on their immunogenicity predicted in step c); [0796] and wherein the polynucleotide sequence of step e) further comprises nucleotide sequences encoding the one or more patient-specific antigens or one or more parts thereof selected in step d).

[0797] 66. A polynucleotide as defined in any of the embodiments B1 to B62.

[0798] 67. A vector comprising the polynucleotide according to embodiment B66.

[0799] 68. A host cell comprising the polynucleotide as defined in any of the embodiments B1 to B62 or comprising the vector according to embodiment B67.

[0800] 69. The polynucleotide according to embodiment B66 formulated for administration to a patient to induce production of a dimeric protein in said patient.

[0801] 70. A polypeptide encoded by the polynucleotide sequence as defined in any of the embodiments B1 to B62.

[0802] 71. A dimeric protein consisting of two polypeptides as defined in embodiment B70.

[0803] 72. The dimeric protein according to embodiment B71, being a homodimeric protein.

[0804] 73. The polynucleotide according to embodiment B66 or the polypeptide according to embodiment B70 or the dimeric protein according to any of embodiments B71 or B72 for use as a medicament.

[0805] 74. A method for preparing an individualized therapeutic anticancer vaccine comprising an immunologically effective amount of a dimeric protein as defined in any of embodiments B1 to B54 or a polypeptide as defined in any of embodiment B1 to B54, the method comprises: [0806] a) transfecting cells with the polynucleotide as defined in any of embodiments B1 to B62; [0807] b) culturing the cells; [0808] c) collecting and purifying the dimeric protein or the polypeptide expressed from the cells; and [0809] d) mixing the dimeric protein or polypeptide obtained from step c) with a pharmaceutically acceptable carrier.

[0810] 75. A method for preparing an individualized therapeutic anticancer vaccine comprising an immunologically effective amount of the polynucleotide as defined in any of embodiments B1 to B62, the method comprises [0811] a) preparing the polynucleotide; [0812] b) optionally cloning the polynucleotide into an expression vector and [0813] c) mixing the polynucleotide from step a) or the vector form step b) with a pharmaceutically acceptable carrier.

[0814] 76. A method of treating a patient having cancer, the method comprising administering to the patient the vaccine according to any of embodiments B1 to B63, which has been prepared specifically for the patient.

[0815] 77. The method according to embodiment B76, wherein the vaccine comprises a polynucleotide and is administered intradermally or intramuscularly.

[0816] 78. The method according to embodiment B77 wherein the polynucleotide is a DNA.

[0817] 79. The method according to embodiment B78 wherein the polynucleotide is an RNA.

[0818] 80. The method according to any of embodiments B76 to B79, wherein administration is carried out with a jet injector.

[0819] 81. The method according to any of embodiments B76 to B80, wherein administration is assisted by electroporation.

[0820] 82. The vaccine according to any of embodiments B1 to B63 for use in a method of treating cancer in a patient, wherein the vaccine has been specifically prepared for the patient.

[0821] 83. Use of the polynucleotide according to embodiment B66 or the polypeptide according to embodiment 7B0 or the dimeric protein according to any of embodiments B71 or B72 for the manufacture of a medicament for the treatment of cancer in a patient, wherein the polynucleotide, polypeptide or dimeric protein has been specifically prepared for the patient.

[0822] 84. A method for preparing the polynucleotide of embodiment B66, said method comprises the steps of: [0823] a) identifying at least one patient-present shared antigen in the tumor tissue or body fluid of a patient [0824] b) determining the patient’s HLA class | and/or class II alleles [0825] c) predicting the immunogenicity of the identified at least one antigen or one or more parts thereof by their predicted binding to the patient’s HLA class | and/or II alleles [0826] d) selecting at least one antigen or one or more parts thereof based on their immunogenicity predicted in step c); [0827] e) preparing a polynucleotide sequence comprising an antigenic unit comprising a nucleotide sequence encoding the at least one antigen or one or more parts thereof selected in step d); and [0828] f) cloning the polynucleotide sequence into an expression vector comprising nucleotide sequences encoding a targeting unit and a dimerization unit.

[0829] 85. The method according to embodiment B84, wherein said method further comprises: [0830] in step a) identifying one or more patient-specific antigens in the tumor tissue of the patient in step c) predicting the immunogenicity of the identified one or more patient-specific antigens or one or more parts thereof by their predicted binding to the patient’s HLA class | and/or II alleles in step d) selecting one or more patient-specific antigens or one or more parts thereof based on their immunogenicity predicted in step c); [0831] and wherein the polynucleotide sequence of step e) further comprises nucleotide sequences encoding the one or more patient-specific antigens or one or more parts thereof selected in step d).