TERT immunogenic compositions and therapeutic methods using the same

Abstract

The present application provides TERT immunogenic compositions and methods of treatment using the same. Disclosed herein are compositions comprising optimized consensus TERT antigens and methods for treating cancer, and in particular immunogenic compositions for treating and providing protection against tumors.

Description

[0001] This application is a divisional application of Chinese patent application No. 201780073893.4, filed on September 29, 2017, entitled “TERT immunogenic composition and treatment method using the same”.

[0002] CROSS-REFERENCE TO RELATED APPLICATIONS

[0003] This application claims priority to U.S. Provisional Application No. 62 / 402,695, filed September 30, 2016, and U.S. Provisional Application No. 62 / 468,124, filed March 7, 2017, which are hereby incorporated by reference in their entireties. Technical Field

[0004] Disclosed herein are compositions and methods for treating cancer, and in particular, immunogenic compositions that treat tumors and provide protection against tumor growth. Background Art

[0005] Cancer is one of the main causes of death in the world, and is the second most common cause of death in the U.S., almost accounting for 1 example in every 4 deaths.Cancer arises from the single cell that has been transformed into tumor cell from normal cell.Such conversion is normally a multi-stage process, progresses to malignant tumor from precancerous lesion.Multiple factors facilitate this progress, including aging, genetic contribution and exposure to external agents such as physical carcinogens (for example, ultraviolet radiation and ionizing radiation), chemical carcinogens (for example, components of asbestos, tobacco smoke etc.) and biological carcinogens (for example, some viruses, bacteria and parasites).

[0006] The prevention, diagnosis and treatment of cancer can take many different forms. Prevention can include screening for predisposing factors (e.g., specific gene variants), changing behavior (e.g., smoking, diet and physical activity), and antiviral (e.g., human papillomavirus, hepatitis B virus) vaccinations. Treatment can include chemotherapy, radiotherapy, and surgical removal of tumors or cancerous tissue. Although many preventive and therapeutic methods are available, such methods generally achieve limited success in effectively preventing and / or treating current cancers.

[0007] Therefore, there is a need to identify and develop compositions and methods for preventing and / or treating cancer to facilitate clinical management of protection against disease progression. In addition, there is a need for more effective treatments to delay disease progression and / or reduce mortality in subjects with cancer. Summary of the Invention

[0008] In one embodiment, the present invention relates to an immunogenic composition comprising a nucleic acid molecule encoding a consensus TERT antigen. In one embodiment, the consensus TERT antigen comprises an amino acid sequence selected from the group consisting of: a) an amino acid sequence selected from the group consisting of SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, SEQ ID NO: 54, and SEQ ID NO: 56, b) an amino acid sequence having 95% or greater identity to an amino acid sequence selected from the group consisting of SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, SEQ ID NO: 54, and SEQ ID NO: 56, and c) a fragment of an amino acid sequence selected from the group consisting of SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, SEQ ID NO: 54, and SEQ ID NO: 56, wherein the fragment comprises at least 95% of the full-length amino acid sequence.

[0009] In one embodiment, the immunogenic composition further comprises one or more nucleotide sequences encoding one or more amino acid sequences selected from the group consisting of: a) an amino acid sequence selected from the group consisting of SEQ ID NO: 2, SEQ ID NO: 4, SEQ ID NO: 6, SEQ ID NO: 8, SEQ ID NO: 10, SEQ ID NO: 12, SEQ ID NO: 14, SEQ ID NO: 16, SEQ ID NO: 18, SEQ ID NO: 20, SEQ ID NO: 22, SEQ ID NO: 24, SEQ ID NO; 58, SEQ ID NO: 60, SEQ ID NO: 62, SEQ ID NO: 64, SEQ ID NO: 66, SEQ ID NO: 68, SEQ ID NO: 70; b) an amino acid sequence having 95% or greater identity to an amino acid sequence selected from the group consisting of SEQ ID NO: 2, SEQ ID NO: 4, SEQ ID NO: 6, SEQ ID NO: 8, SEQ ID NO: 10, SEQ ID NO: 12, SEQ ID NO: 14, SEQ ID NO: 16, SEQ ID NO: 18, SEQ ID NO: 20, SEQ ID NO: 22, SEQ ID NO: 24, SEQ ID NO; 58, SEQ ID NO: 60, SEQ ID NO: 62, SEQ ID NO: 64, SEQ ID NO: 66, SEQ ID NO: 68, SEQ ID NO: 70 NO:20, SEQ ID NO:22, SEQ ID NO:24, SEQ ID NO;58, SEQ ID NO:60, SEQ ID NO:62, SEQ ID NO:64, SEQ ID NO:66, SEQ ID NO:68, SEQ ID NO:70, and c) a fragment of an amino acid sequence selected from the group consisting of SEQ ID NO:2, SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:8, SEQ ID NO:10, SEQ ID NO:12, SEQ ID NO:14, SEQ ID NO:16, SEQ ID NO:18, SEQ ID NO:20, SEQ ID NO:22, SEQ ID NO:24, SEQ ID NO;58, SEQ ID NO:60, SEQ ID NO:62, SEQ ID NO:64, SEQ ID NO:66, SEQ ID NO:68, SEQ ID NO:70, wherein the fragment comprises at least 95% of the full-length amino acid sequence.

[0010] In one embodiment, the immunogenic composition further comprises a nucleotide sequence encoding one or more antigens selected from the group consisting of: MAGE A1, gp100, viral antigens, and combinations thereof. In one embodiment, the viral antigen is an antigen from hepatitis B virus (HBV), hepatitis C virus (HCV), or human papillomavirus (HPV). In one embodiment, the HBV antigen is an HBV core antigen or an HBV surface antigen, or a combination thereof. In one embodiment, the HCV antigen is an HCV NS34A antigen, an HCV NS5A antigen, an HCV NS5B antigen, an HCV NS4B antigen, or a combination thereof. In one embodiment, the HPV antigen is an HPV 6 type E6 antigen, an HPV 6 type E7 antigen, an HPV 11 type E6 antigen, an HPV 11 type E7 antigen, an HPV 16 type E6 antigen, an HPV 16 type E7 antigen, an HPV 18 type E6 antigen, an HPV 18 type E7 antigen, or a combination thereof.

[0011] In one embodiment, the immunogenic composition further comprises an immune checkpoint inhibitor.

[0012] In one embodiment, the nucleic acid molecule comprises a nucleotide sequence selected from the group consisting of: a) a nucleotide sequence selected from the group consisting of: SEQ ID NO:45, SEQ ID NO:47, SEQ ID NO:49, SEQ ID NO:51, SEQ ID NO:53, and SEQ ID NO:55, b) a nucleotide sequence having 95% or greater identity to a nucleotide sequence selected from the group consisting of: SEQ ID NO:45, SEQ ID NO:47, SEQ ID NO:49, SEQ ID NO:51, SEQ ID NO:53, and SEQ ID NO:55, and c) a fragment of a nucleotide sequence selected from the group consisting of: SEQ ID NO:45, SEQ ID NO:47, SEQ ID NO:49, SEQ ID NO:51, SEQ ID NO:53, and SEQ ID NO:55, wherein the fragment comprises at least 95% of the full-length nucleotide sequence.

[0013] In one embodiment, the immunogenic composition comprises one or more nucleotide sequences selected from the group consisting of: a) a nucleotide sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 3, SEQ ID NO: 5, SEQ ID NO: 7, SEQ ID NO: 9, SEQ ID NO: 11, SEQ ID NO: 13, SEQ ID NO: 15, SEQ ID NO: 17, SEQ ID NO: 19, SEQ ID NO: 21, SEQ ID NO: 23, SEQ ID NO: 57, SEQ ID NO: 59, SEQ ID NO: 61, SEQ ID NO: 63, SEQ ID NO: 65, SEQ ID NO: 67, and SEQ ID NO: 69; b) a nucleotide sequence having 95% or greater identity to a nucleotide sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 3, SEQ ID NO: 5, SEQ ID NO: 7, SEQ ID NO: 9, SEQ ID NO: 11, SEQ ID NO: 13, SEQ ID NO: 15, SEQ ID NO: 17, SEQ ID NO: 19, SEQ ID NO: 21, SEQ ID NO: 23, SEQ ID NO: 57, SEQ ID NO: 59, SEQ ID NO: 61, SEQ ID NO: 63, SEQ ID NO: 65, SEQ ID NO: 67, and SEQ ID NO: 69; NO:21, SEQ ID NO:23, SEQ ID NO:57, SEQ ID NO:59, SEQ ID NO:61, SEQ ID NO:63, SEQ ID NO:65, SEQ ID NO:67 and SEQ ID NO:69, and c) a fragment of a nucleotide sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:5, SEQ ID NO:7, SEQ ID NO:9, SEQ ID NO:11, SEQ ID NO:13, SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO:19, SEQ ID NO:21, SEQ ID NO:23, SEQ ID NO:57, SEQ ID NO:59, SEQ ID NO:61, SEQ ID NO:63, SEQ ID NO:65, SEQ ID NO:67 and SEQ ID NO:69, wherein the fragment comprises at least 95% of the full-length nucleotide sequence.

[0014] In one embodiment, the nucleic acid molecule is a plasmid.

[0015] In one embodiment, the immunogenic composition comprises one or more plasmids.

[0016] In one embodiment, the immunogenic composition further comprises an adjuvant. In one embodiment, the adjuvant is IL-12, IL-15, IL-28 or RANTES.

[0017] In one embodiment, the present invention relates to a method for treating or preventing cancer in a subject in need thereof, the method comprising administering to the subject an immunogenic composition comprising a nucleic acid molecule encoding a common TERT antigen. In one embodiment, the administration method comprises an electroporation step. In this embodiment, the method further comprises administering to the subject an immune checkpoint inhibitor.

[0018] In one embodiment, the immunogenic composition and the immune checkpoint inhibitor are administered to the subject in a single formulation. In one embodiment, the immunogenic composition and the immune checkpoint inhibitor are administered to the subject separately.

[0019] In one embodiment, the cancer is selected from the group consisting of melanoma, head and neck cancer, prostate cancer, liver cancer, cervical cancer, recurrent respiratory papillomatosis (RRP), anal cancer, blood cancer, ovarian cancer, and combinations thereof.

[0020] In one embodiment, the present invention relates to a nucleic acid molecule comprising one or more nucleotide sequences selected from the group consisting of: a) a nucleotide sequence selected from the group consisting of: SEQ ID NO:45, SEQ ID NO:47, SEQ ID NO:49, SEQ ID NO:51, SEQ ID NO:53 and SEQ ID NO:55, b) a nucleotide sequence having 95% or greater identity to a nucleotide sequence selected from the group consisting of: SEQ ID NO:45, SEQ ID NO:47, SEQ ID NO:49, SEQ ID NO:51, SEQ ID NO:53 and SEQ ID NO:55, and c) a fragment of a nucleotide sequence selected from the group consisting of: SEQ ID NO:45, SEQ ID NO:47, SEQ ID NO:49, SEQ ID NO:51, SEQ ID NO:53 and SEQ ID NO:55, wherein the fragment comprises at least 95% of the full-length nucleotide sequence.

[0021] In one embodiment, the nucleic acid molecule further comprises one or more nucleotide sequences selected from the group consisting of: a) a nucleotide sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 3, SEQ ID NO: 5, SEQ ID NO: 7, SEQ ID NO: 9, SEQ ID NO: 11, SEQ ID NO: 13, SEQ ID NO: 15, SEQ ID NO: 17, SEQ ID NO: 19, SEQ ID NO: 21, SEQ ID NO: 23, SEQ ID NO: 57, SEQ ID NO: 59, SEQ ID NO: 61, SEQ ID NO: 63, SEQ ID NO: 65, SEQ ID NO: 67, and SEQ ID NO: 69; b) a nucleotide sequence having 95% or greater identity to a nucleotide sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 3, SEQ ID NO: 5, SEQ ID NO: 7, SEQ ID NO: 9, SEQ ID NO: 11, SEQ ID NO: 13, SEQ ID NO: 15, SEQ ID NO: 17, SEQ ID NO: 19, SEQ ID NO: 21, SEQ ID NO: 23, SEQ ID NO: 57, SEQ ID NO: 59, SEQ ID NO: 61, SEQ ID NO: 63, SEQ ID NO: 65, SEQ ID NO: 67, and SEQ ID NO: 69; NO:21, SEQ ID NO:23, SEQ ID NO:57, SEQ ID NO:59, SEQ ID NO:61, SEQ ID NO:63, SEQ ID NO:65, SEQ ID NO:67 and SEQ ID NO:69, and c) a fragment of a nucleotide sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:5, SEQ ID NO:7, SEQ ID NO:9, SEQ ID NO:11, SEQ ID NO:13, SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO:19, SEQ ID NO:21, SEQ ID NO:23, SEQ ID NO:57, SEQ ID NO:59, SEQ ID NO:61, SEQ ID NO:63, SEQ ID NO:65, SEQ ID NO:67 and SEQ ID NO:69, wherein the fragment comprises at least 95% of the full-length nucleotide sequence.

[0022] In one embodiment, the nucleic acid molecule is a plasmid.

[0023] In one embodiment, the present invention relates to an amino acid molecule comprising one or more amino acid sequences selected from the group consisting of: a) an amino acid sequence selected from the group consisting of SEQ ID NO:46, SEQ ID NO:48, SEQ ID NO:50, SEQ ID NO:52, SEQ ID NO:54 and SEQ ID NO:56, b) an amino acid sequence having 95% or greater identity to an amino acid sequence selected from the group consisting of SEQ ID NO:46, SEQ ID NO:48, SEQ ID NO:50, SEQ ID NO:52, SEQ ID NO:54 and SEQ ID NO:56, and c) a fragment of an amino acid sequence selected from the group consisting of SEQ ID NO:46, SEQ ID NO:48, SEQ ID NO:50, SEQ ID NO:52, SEQ ID NO:54 and SEQ ID NO:56, wherein the fragment comprises at least 95% of the full-length amino acid sequence.

[0024] In one embodiment, the amino acid molecule further comprises one or more amino acid sequences selected from the group consisting of: a) an amino acid sequence selected from the group consisting of SEQ ID NO: 2, SEQ ID NO: 4, SEQ ID NO: 6, SEQ ID NO: 8, SEQ ID NO: 10, SEQ ID NO: 12, SEQ ID NO: 14, SEQ ID NO: 16, SEQ ID NO: 18, SEQ ID NO: 20, SEQ ID NO: 22, SEQ ID NO: 24, SEQ ID NO: 58, SEQ ID NO: 60, SEQ ID NO: 62, SEQ ID NO: 64, SEQ ID NO: 66, SEQ ID NO: 68, SEQ ID NO: 70; b) an amino acid sequence having 95% or greater identity to an amino acid sequence selected from the group consisting of SEQ ID NO: 2, SEQ ID NO: 4, SEQ ID NO: 6, SEQ ID NO: 8, SEQ ID NO: 10, SEQ ID NO: 12, SEQ ID NO: 14, SEQ ID NO: 16, SEQ ID NO: 18, SEQ ID NO: 20, SEQ ID NO: 22, SEQ ID NO: 24, SEQ ID NO: 58, SEQ ID NO: 60, SEQ ID NO: 62, SEQ ID NO: 64, SEQ ID NO: 66, SEQ ID NO: 68, SEQ ID NO: 70 NO:22, SEQ ID NO:24, SEQ ID NO;58, SEQ ID NO:60, SEQ ID NO:62, SEQ ID NO:64, SEQ ID NO:66, SEQ ID NO:68, SEQ ID NO:70, and c) a fragment of an amino acid sequence selected from the group consisting of SEQ ID NO:2, SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:8, SEQ ID NO:10, SEQ ID NO:12, SEQ ID NO:14, SEQ ID NO:16, SEQ ID NO:18, SEQ ID NO:20, SEQ ID NO:22, SEQ ID NO:24, SEQ ID NO;58, SEQ ID NO:60, SEQ ID NO:62, SEQ ID NO:64, SEQ ID NO:66, SEQ ID NO:68, SEQ ID NO:70, wherein the fragment comprises at least 95% of the full-length amino acid sequence. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] Figure 1A to Figure 1E depict the construction of pTyr.

[0026] Figures 2A-2B depict experimental results showing, respectively, the immunization strategy and the induction of a cell-mediated immune response by Tyr DNA vaccination.

[0027] Figure 3 Depicted are experimental results showing flow cytometric fluorescence activated cell sorting (FACS) of control and immunized mice.

[0028] Figures 4A-4B depict experimental results showing the induction of tyrosinase-specific antibodies in immunized mice.

[0029] Figures 5A-5B depict experimental results showing Kaplan-Meier survival curves and tumor volume curves after tumor challenge in control mice and immunized mice, respectively.

[0030] Figures 6A to 6B , depicts experimental results showing MDSC cell populations in immunized and non-immunized mice.

[0031] Figure 7 Depicted are the results of an experiment showing staining for MDSCs in mice immunized with pVax1 and pTyr.

[0032] 8A-8B depict the results of experiments showing MCP-1 secretion by MDSCs.

[0033] Figure 9 Depicted are experimental results showing the phylogenetic relationships of Tyr nucleotide sequences among the indicated organisms.

[0034] Figure 10A 10C , depicted are experimental results showing: (A) a schematic diagram showing the plasmid map of pPRAME (also referred to herein as pGX1411); (B) staining of nuclei of RD and 293T cells and staining for the consensus PRAME antigen using DAPI; and (C) western blot of the consensus PRAME antigen in lysates from untransfected cells ("control"), cells transfected with pVAX ("pVAX"), and cells transfected with pPRAME ("PRAME-pVAX").

[0035] Figures 11A-11B depict the results of an experiment showing that groups of mice were treated with plaque forming units (SFU) / 10 against interferon gamma (IFN-γ). 6 Results of the splenocyte mapping experiment.

[0036] Figure 12A12C , experimental results showing: (A) a schematic diagram showing the plasmid map of pNY-ESO-1 (also referred to herein as pGX1409); (B) staining of cell nuclei and consensus NY-ESO-1 antigen with DAPI; and (C) western blots of consensus NY-ESO-1 antigen in RD and 293T lysates from untransfected cells ("control"), cells transfected with pVAX ("pVAX"), and cells transfected with pNY-ESO-1 ("pNY-ESO-1").

[0037] Figure 13 Figure 3. Groups of mice were treated with plaque forming units (SFU) / 10 against interferon gamma (IFN-γ). 6 Figure 3. A graph showing the mapping of spleen cells.

[0038] Figure 14 Figure 3. Groups of mice were treated with plaque forming units (SFU) / 10 against interferon gamma (IFN-γ). 6 Figure 3. A graph showing the mapping of spleen cells.

[0039] Figure 15 Depicted is a schematic diagram showing various cancers and some of their associated cancer antigens.

[0040] Figure 16 Schematic design of TERT constructs is depicted. A synthetic consensus hTERT was designed for human TERT, harboring five mutations that abolish its function. Native RhTERT encodes rhesus macaque TERT with two mutations. Both constructs were optimized. The table shows the protein homology between the optimized TERT and native TERT identified in humans, rhesus macaques, and mice.

[0041] Figure 17A to Figure 17B , depicts characterization of mouse TERT-specific IFN-γ responses to rhesus monkey native peptides. ( FIG17A ) Immunization schedule. Mice (n=8 / group) were immunized with native RhTERT and synthetic consensus hTERT via IM / electroporation at weeks 0, 2, and 4. ( Figure 17B ) The frequency of TERT-specific IFN-γ plaque-forming units (SFU) per million splenocytes isolated from vaccinated mice was determined by IFN-γ ELISpot assay using rhesus monkey peptide.

[0042] 18A to 18C , depicts the characterization of TERT-specific IFN-γ responses in rhesus macaques. ( Figure 18AImmunization schedule. Rhesus macaques (n=5 / group) were immunized with native RhTERT or synthetic consensus hTERT and rhIL-12 by IM / electroporation at weeks 0, 4, 8, and 12. (Figure 18B and Figure 18C ) The frequency of TERT-specific IFN-γ plaque-forming units (SFU) per million splenocytes isolated from vaccinated mice was determined by IFN-γ ELISpot assay using native rhesus peptide and synthetic consensus hTERT peptide.

[0043] Figure 19 Depicts experimental results showing that synthetic consensus hTERT (pGX1434) induces a strong T cell response and slows TC-1 tumor growth in mice. (Left) Mice received three immunizations (with 0 μg, 20 μg, 40 μg, or 60 μg of pGX1434) separated by 2 weeks. T cell responses in mice were assessed at 5 weeks, showing that pGX1434 induces a strong T cell response. (Right) Mice were implanted with 25 x 10 4 TC-1 cells. They were then immunized four times with 25 μg of pGX1434, one week apart. Tumor volume was measured over 32 days, showing that mice treated with pGX1434 had reduced tumor volume compared to naive mice.

[0044] Figure 20 Depicted are the results of experiments showing the immunogenicity of SynCon hTERT (pGX1434) in mice.

[0045] Figure 21A to Figure 21B , depicts experimental results showing that SyCon hTERT (pGX1434) and mut-hTERT (pGX1406) provide similar control of TC-1 tumor growth.

[0046] Figure 22A to Figure 22B , depicts the experimental design for experiments showing the immunogenicity of hTERT in non-human primates (NHPs). Figure 22A depicts an exemplary immunization schedule. Figure 22B A table showing the constructs administered to the different groups is depicted: pGX1434: hTERT; pGX1406: Mut-hTERT; pGX1447: Mut-rhTERT; pGX1404: WT-1; pGX1108: PSMA; pGX6006: rhIL-12.

[0047] Figure 23A to Figure 23B, depicts the results of experiments showing that SynCon hTERT breaks self-tolerance in NHPs. Figure 23A depicts the results of experiments showing that responses to native rhesus TERT were detected at PD3 and PD4 in Group 1 animals. Figure 23B Depicted are the results of experiments showing that responses to native rhesus TERT were detected at PD3 and PD4 in Group 2 animals.

[0048] Figure 24A to Figure 24C , depicts experimental results showing that SynCon design enables breaking of self-tolerance. Figure 24A depicts a table showing the percent identity of different constructs to the native sequence. Figure 24B depicts experimental results showing TERT-specific IFNγ responses. Figure 24C Depicted are the results of experiments demonstrating TERT-specific IFNγ responses in rhesus macaques.

[0049] Figure 25A to Figure 25B , depicts the results of experiments showing the NHP immunogenicity of SynCon hTERT. FIG25A depicts the results of experiments showing the TERT response generated using a combination of SynCon hTERT, PSMA, WT-1, and IL-12. Figure 25B Depicted are the results of experiments showing TERT responses generated using a combination of mut-hTERT, PSMA, WT-1, and IL-12.

[0050] Figure 26A to Figure 26C , depicts experimental results showing the NHP immunogenicity of SynCon hTERT. FIG26A depicts experimental results showing the TERT response generated using a combination of TERT, PSMA, WT-1, and IL-12. FIG26B depicts experimental results showing the PSMA response generated using a combination of TERT, PSMA, WT-1, and IL-12. Figure 26C Depicted are the results of experiments showing WT-1 responses using a combination of TERT, PSMA, WT-1, and IL-12.

[0051] Figure 27A to Figure 27B , depicts experimental results showing the breadth of the multivalent SynCon vaccine. Figure 27A depicts the immunization schedule used. Figure 27B The percentage of antigen-specific T cells identified after immunization is depicted.

[0052] Figure 28A to Figure 28B , depicts experimental results showing that multivalent SynCon vaccines reduce tumor burden and increase survival. Figure 28A depicts the immunization schedule used. Figure 28BDepicted are the mean tumor volume and % survival of animals immunized with a combination of WT1, PSMA, and SynCon mTERT.

[0053] Details

[0054] The present invention relates to an immunogenic composition comprising a synthetic consensus TERT antigen alone or in combination with an inhibitor of at least one other cancer antigen or immune checkpoint protein, and a method for treating a disease or condition using the composition. In one embodiment, the immunogenic composition comprises TERT and at least one other cancer antigen consensus sequence. The cancer antigen consensus sequence that may be included in the immunogenic composition includes but is not limited to tyrosinase (Tyr) (antigen preferentially expressed in melanoma (PRAME)), tyrosinase-related protein 1 (Tyrp1), cancer testis antigen (NY-ESO-1), hepatitis B virus antigen, prostate specific antigen (PSA), prostate specific membrane antigen (PSMA), prostate six transmembrane epithelial antigen (STEAP), prostate stem cell antigen (PSCA), fibroblast activation protein (FAP), follicle stimulating hormone receptor (FSHR) and Wilms tumor 1 antigen (WT-1). In one embodiment, the immunogenic composition of the present invention can provide a combination of cancer antigens for preventing or treating cancer in a subject in need.

[0055] One way to design nucleic acids and corresponding encoded amino acid sequences of recombinant cancer antigens is to introduce mutations that change specific amino acids in the complete amino acid sequence of the natural cancer antigen. The introduction of mutations will not change the cancer antigen so much that it cannot be universally administered across mammalian subjects (e.g., human or dog subjects), but will change the antigen enough to break tolerance or be treated as an exogenous antigen to produce an immune response. Another way can be to produce a common recombinant cancer antigen with at least 85% and up to 99% amino acid sequence identity, at least 90% and up to 98% sequence identity, at least 93% and up to 98% sequence identity, or at least 95% and up to 98% sequence identity with the corresponding natural cancer antigen. In some cases, the recombinant cancer antigen has 95%, 96%, 97%, 98% or 99% amino acid sequence identity with the corresponding natural cancer antigen. The natural cancer antigen is an antigen that is generally associated with a specific cancer or cancer tumor. Depending on the cancer antigen, the consensus sequence of the cancer antigen can be across mammalian species or within a subtype of a species or across viral strains or serotypes. Some cancer antigens will not change greatly with the wild-type amino acid sequence of cancer antigens. Some cancer antigens have nucleic acid / amino acid sequences with such large differences that they can not produce consensus sequences between species. In these cases, recombinant cancer antigens that can break tolerance and produce an immune response are produced, and the recombinant cancer antigens have at least 85% and up to 99% amino acid sequence identity, at least 90% and up to 98% sequence identity, at least 93% and up to 98% sequence identity, or at least 95% and up to 98% sequence identity with the corresponding natural cancer antigen. In some cases, the recombinant cancer antigens have 95%, 96%, 97%, 98% or 99% amino acid sequence identity with the corresponding natural cancer antigen. The aforementioned methods can be combined so that the final recombinant cancer antigen has the percentage similarity with the natural cancer antigen amino acid sequence as discussed above.

[0056] The recombinant cancer antigen can induce antigen-specific T cells and / or high-titer antibody responses, thereby inducing or triggering an immune response against a cancer or tumor expressing the antigen or reacting with the cancer or tumor. In some embodiments, the immune response induced or triggered can be a cellular, humoral, or cellular and humoral immune response. In some embodiments, the cellular immune response induced or triggered may include the induction or secretion of interferon-γ (IFN-γ) and / or tumor necrosis factor α (TNF-α). In other embodiments, the immune response induced or triggered can reduce or inhibit one or more immunosuppressive factors that promote the growth of tumors or cancers expressing the antigen, such as, but not limited to, factors that downregulate MHC presentation, factors that upregulate antigen-specific regulatory T cells (Treg), PD-L1, FasL, cytokines such as IL-10 and TFG-β, tumor-associated macrophages, tumor-associated fibroblasts, soluble factors produced by immunosuppressive cells, CTLA-4, PD-1, MDSC, MCP-1, and immune checkpoint molecules.

[0057] Immunogenic compositions can be further combined with antibodies for checkpoint inhibitors such as PD-1 and PDL-1 to enhance stimulation of cellular and humoral immune responses. Anti-PD-1 or anti-PDL-1 antibodies are used to prevent PD-1 or PDL-1 from suppressing T cells and / or B cell responses. In general, by designing cancer antigens to be recognized by the immune system, it is possible to help overcome other forms of immunosuppression produced by tumor cells, and these immunogenic compositions can be used in combination with suppression or inhibitory therapy (such as anti-PD-1 and anti-PDL-1 antibody therapy) to further enhance T cells and / or B cell responses.

[0058] 1. Definition

[0059] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those generally understood by those of ordinary skill in the art. In the event of conflict, this document (including definitions) shall prevail. Exemplary methods and materials are described below, although methods and materials similar or equivalent to those described herein can be used in the implementation or testing of the present invention. All publications, patent applications, patents and other references mentioned herein are incorporated by reference in their entirety. The materials, methods and examples disclosed herein are merely illustrative and are not intended to be limiting. The terms used herein are only for the purpose of describing specific embodiments and are not intended to be limiting.

[0060] As used herein, the terms "comprise," "include," "have," "have," "may," "contain," and variations thereof are intended to be open transitional phrases, terms, or words that do not exclude the possibility of additional actions or structures. The singular forms "a," "and," and "the" include plural references unless the context clearly dictates otherwise. The present disclosure also encompasses other embodiments that "comprise," "consist of," and "consist essentially of" the embodiments or components set forth herein, whether or not explicitly stated.

[0061] For the recitation of numerical ranges herein, every intervening number therebetween is expressly included with equal precision. For example, for the range 6-9, the numbers 7 and 8 are included in addition to 6 and 9, and for the range 6.0-7.0, the numbers 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, and 7.0 are expressly included.

[0062] As used herein, "adjuvant" means any molecule added to the immunogenic compositions described herein to enhance the immunogenicity of the antigens encoded by the nucleic acid molecules and encoding nucleic acid sequences described below.

[0063] As used herein, "antibody" means an antibody of the class IgG, IgM, IgA, IgD or IgE, or a fragment or derivative thereof, including Fab, F(ab')2, Fd, and single-chain antibodies, diabodies, bispecific antibodies, bifunctional antibodies, and derivatives thereof. The antibody can be an antibody isolated from a serum sample of a mammal, a polyclonal antibody, an affinity-purified antibody, or a mixture thereof, which exhibits sufficient binding specificity to a desired epitope or a sequence derived therefrom.

[0064] As used herein, "coding sequence" or "coding nucleic acid" means a nucleic acid (RNA or DNA molecule) comprising a nucleotide sequence that encodes a protein. The coding sequence may also include start and stop signals operably linked to regulatory elements (including a promoter and polyadenylation signals) capable of directing expression in the cells of an individual or mammal to which the nucleic acid is administered.

[0065] As used herein, "complementary" or "complementary" refers to nucleic acids and can refer to Watson-Crick (eg, AT / U and CG) or Hoogsteen base pairing between nucleotides or nucleotide analogs of a nucleic acid molecule.

[0066] As used herein, "consensus" or "consensus sequence" refers to a polypeptide sequence based on an analysis of an alignment of multiple sequences of the same gene from different organisms. Nucleic acid sequences encoding the consensus polypeptide sequence can be prepared. Immunogenic compositions comprising proteins containing the consensus sequence and / or nucleic acids encoding such proteins can be used to induce broad immunity against the antigen.

[0067] "Electroporation," "electroperization," or "electrokinetic enhancement" ("EP"), as used interchangeably herein, refers to the use of transmembrane electric field pulses to induce microscopic pathways (pores) in biological membranes; their presence allows the passage of biomolecules such as plasmids, oligonucleotides, siRNA, drugs, ions, and water from one side of the cell membrane to the other.

[0068] As used herein with respect to nucleic acid sequences, "fragment" means a nucleic acid sequence or portion thereof that encodes a polypeptide capable of eliciting an immune response in a mammal that cross-reacts with an antigen disclosed herein. The fragment can be a DNA fragment selected from at least one of the various nucleotide sequences encoding protein fragments described below. A fragment can comprise at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 95% of one or more nucleic acid sequences described below. In some embodiments, a fragment may comprise at least 20 nucleotides or more, at least 30 nucleotides or more, at least 40 nucleotides or more, at least 50 nucleotides or more, at least 60 nucleotides or more, at least 70 nucleotides or more, at least 80 nucleotides or more, at least 90 nucleotides or more, at least 100 nucleotides or more, at least 150 nucleotides or more, at least 200 nucleotides or more, at least 250 nucleotides or more, at least 300 nucleotides or more, at least 350 nucleotides or more, at least 400 nucleotides or more, at least 450 nucleotides or more, at least 500 nucleotides or more, at least 550 nucleotides or more, at least 600 nucleotides or more, at least 650 nucleotides or more, at least 700 nucleotides or more, at least 750 nucleotides or more, at least 800 nucleotides or more, at least 850 nucleotides or more, at least 900 nucleotides or more, at least 950 nucleotides or more, or at least 1000 nucleotides or more of at least one of the nucleic acid sequences described below.

[0069] With respect to polypeptide sequences, "fragment" or "immunogenic fragment" means a polypeptide capable of eliciting an immune response in a mammal that cross-reacts with an antigen disclosed herein. A fragment can be a polypeptide fragment selected from at least one of the various amino acid sequences described below. A fragment of a consensus protein can comprise at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, or at least 95% of the consensus protein. In some embodiments, a fragment of a consensus protein may comprise at least 20 amino acids or more, at least 30 amino acids or more, at least 40 amino acids or more, at least 50 amino acids or more, at least 60 amino acids or more, at least 70 amino acids or more, at least 80 amino acids or more, at least 90 amino acids or more, at least 100 amino acids or more, at least 110 amino acids or more, at least 120 amino acids or more, at least 130 amino acids or more, at least 140 amino acids or more, at least 150 amino acids or more, at least 160 amino acids or more, at least 170 amino acids or more, at least 180 amino acids or more amino acids of a protein sequence disclosed herein.

[0070] As used herein, "genetic construct" refers to a DNA or RNA molecule comprising a nucleotide sequence encoding a protein. The coding sequence includes start and stop signals operably linked to regulatory elements (including a promoter and a polyadenylation signal) that can direct expression in the cells of an individual to whom the nucleic acid molecule is administered. As used herein, the term "expressible form" refers to a genetic construct containing the necessary regulatory elements operably linked to the coding sequence encoding the protein so that when present in the cells of an individual, the coding sequence will be expressed.

[0071] As used herein, the term "homology" refers to the degree of complementarity. Partial homology or complete homology (i.e., identity) may exist. A partially complementary sequence that at least partially inhibits hybridization of a completely complementary sequence to a target nucleic acid refers to the use of the functional term "substantially homologous". When used to refer to double-stranded nucleic acid sequences such as cDNA or genomic clones, as used herein, the term "substantially homologous" refers to probes that can hybridize to the chains of a double-stranded nucleic acid sequence under conditions of low stringency. When used to refer to a single-stranded nucleic acid sequence, as used herein, the term "substantially homologous" refers to probes that can hybridize to (i.e., be complementary to) a single-stranded nucleic acid template sequence under conditions of low stringency.

[0072] As used herein in the context of two or more nucleic acid or polypeptide sequences, "identical" or "identity" means that a sequence has a specified percentage of residues that are identical over a specified region. Percentages can be calculated as follows: the two sequences are optimally aligned, the two sequences are compared over a specified region, the number of positions at which identical residues are present in the two sequences is determined to obtain the number of matching positions, the number of matching positions is divided by the total number of positions in the specified region, and the result is multiplied by 100 to obtain the percentage of sequence identity. Where the two sequences have different lengths or the alignment produces one or more staggered ends and the specified comparison region only includes a single sequence, the residues of the single sequence are included in the denominator of the calculation rather than the numerator. When comparing DNA and RNA, thymine (T) and uracil (U) can be considered to be equivalent. Identity can be obtained artificially or by using a computer sequence algorithm such as BLAST or BLAST 2.0.

[0073] As used herein, "immune response" means the activation of a host's immune system, such as a mammal's immune system, in response to the introduction of an antigen. The immune response can be in the form of a cellular response or a humoral response, or both.

[0074] As used herein, "nucleic acid" or "oligonucleotide" or "polynucleotide" means at least two nucleotides covalently linked together. The description of a single strand also defines the sequence of the complementary strand. Thus, nucleic acid also includes the complementary strand of the described single strand. Many variants of a nucleic acid can be used for the same purpose as a given nucleic acid. Thus, nucleic acid also includes substantially identical nucleic acids and their complementary sequences. A single strand provides a probe that can hybridize to a target sequence under stringent hybridization conditions. Thus, nucleic acid also includes probes that hybridize under stringent hybridization conditions.

[0075] Nucleic acids can be single-stranded or double-stranded, or can contain portions of double-stranded and single-stranded sequences. Nucleic acids can be DNA (genomic DNA and cDNA), RNA, or hybrids, wherein nucleic acids can contain a combination of deoxyribonucleotides and ribonucleotides, and a combination of bases (including uracil, adenine, thymine, cytosine, guanine, inosine, xanthine, hypoxanthine, isocytosine, and isoguanine). Nucleic acids can be obtained by chemical synthesis or by recombinant methods.

[0076] As used herein, "operably linked" means that the expression of a gene is under the control of a promoter to which it is spatially linked. A promoter can be located 5' (upstream) or 3' (downstream) of the gene under its control. The distance between the promoter and the gene can be approximately the same as the distance between the promoter and the gene it controls in the gene from which the promoter is derived. As is known in the art, variations in this distance are acceptable without loss of promoter function.

[0077] As used herein, "peptide," "protein," or "polypeptide" may mean a linked sequence of amino acids, and may be natural, synthetic, or modified, or a combination of natural and synthetic.

[0078] As used herein, "promoter" means a molecule of synthetic or natural origin that is capable of conferring, activating or enhancing the expression of a nucleic acid in a cell. A promoter may comprise one or more specific transcriptional regulatory sequences to further enhance the expression of the sequence and / or alter its spatial expression and / or temporal expression. A promoter may also comprise distal enhancer or repressor elements that are up to several thousand base pairs away from the transcription start site. Promoters may be derived from sources including viruses, bacteria, fungi, plants, insects and animals. A promoter may constitutively or differentially regulate the expression of a genomic component for the cell, tissue or organ where expression occurs or for the developmental stage at which expression occurs, or in response to external stimuli such as physiological stress, pathogens, metal ions or inducers. Representative examples of promoters include bacteriophage T7 promoter, bacteriophage T3 promoter, SP6 promoter, lac operator-promoter, tac promoter, SV40 late promoter, SV40 early promoter, RSV-LTR promoter, CMV IE promoter, SV40 early promoter or SV 40 late promoter and CMV IE promoter.

[0079] "Signal peptide" and "leader sequence" are used interchangeably herein and refer to an amino acid sequence that can be attached to the amino terminus of a protein described herein. The signal peptide / leader sequence generally directs the localization of the protein. As used herein, the signal peptide / leader sequence can promote the secretion of a protein from the cell in which it is produced. After secretion from the cell, the signal peptide / leader sequence is typically cleaved from the remainder of the protein (usually referred to as the mature protein). The signal peptide / leader sequence is attached to the amino terminus (i.e., the N terminus) of the protein.

[0080] As used herein, "stringent hybridization conditions" means conditions under which a first nucleic acid sequence (e.g., a probe) will hybridize to a second nucleic acid sequence (e.g., a target) (such as in a complex mixture of nucleic acids). Stringent conditions are sequence-dependent and are different in different circumstances. Stringent conditions can be selected to be about 5-10°C lower than the thermal melting point (Tm) of the specific sequence at a defined ionic strength pH. The Tm can be the temperature (under defined ionic strength, pH, and nucleic acid concentration) at which 50% of the probes complementary to the target hybridize to the target sequence at equilibrium (because the target sequence is present in excess, 50% of the probes are occupied at equilibrium at the Tm). Stringent conditions can be those conditions where the salt concentration is less than about 1.0 M sodium ion, such as a sodium ion concentration of about 0.01-1.0 M (or other salts) at pH 7.0 to 8.3 and the temperature is at least about 30°C for short probes (e.g., about 10-50 nucleotides) and at least about 60°C for long probes (e.g., greater than about 50 nucleotides). Stringent conditions can also be achieved by adding destabilizing agents such as formamide. For selective or specific hybridization, a positive signal can be at least 2 to 10 times the background hybridization. Exemplary stringent hybridization conditions include the following: 50% formamide, 5x SSC and 1% SDS, incubated at 42°C, or 5x SSC, 1% SDS, incubated at 65°C, washed in 0.2x SSC and 0.1% SDS at 65°C.

[0081] As used herein, "subject" may refer to a mammal that is desired or required to be immunized with the immunogenic composition described herein. The mammal may be a human, chimpanzee, dog, cat, horse, cow, mouse, or rat.

[0082] As used herein, "substantially complementary" means that a first sequence is substantially complementary to the complement of a second sequence within 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 180, 270, 360, In some embodiments, the two sequences are at least 60%, 65%, 70%, 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical over a region of 450, 540 or more nucleotides or amino acids, or the two sequences hybridize under stringent hybridization conditions.

[0083] As used herein, "substantially identical" means that if the first sequence is substantially complementary to the complement of the second sequence, then the first and second sequences are substantially complementary. The polypeptides are at least 60%, 65%, 70%, 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical over a region or with respect to a nucleic acid of 100, 180, 270, 360, 450, 540 or more nucleotides or amino acids.

[0084] As used herein, "treating" or "treating" may mean protecting an animal from a disease by preventing, inhibiting, suppressing, or completely eliminating the disease. Preventing a disease includes administering an immunogenic composition of the present invention to an animal prior to the onset of the disease. Suppressing a disease includes administering an immunogenic composition of the present invention to an animal after the disease has been initiated but before its clinical manifestation. Suppressing a disease includes administering an immunogenic composition of the present invention to an animal after clinical manifestation of the disease.

[0085] As used herein, "variant" with respect to a nucleic acid means (i) a portion or fragment of a reference nucleotide sequence; (ii) the complement of a reference nucleotide sequence or a portion thereof; (iii) a nucleic acid substantially identical to a reference nucleic acid or its complement; or (iv) a nucleic acid that hybridizes under stringent conditions to a reference nucleic acid, its complement, or a sequence substantially identical thereto.

[0086] For peptides or polypeptides, "variant" refers to a peptide or polypeptide that differs in amino acid sequence by insertion, deletion, or conservative substitution of amino acids, but retains at least one biological activity. Variant can also refer to a protein having an amino acid sequence that is substantially identical to a reference protein having an amino acid sequence that retains at least one biological activity. Conservative substitution of amino acids, i.e., replacing an amino acid with a different amino acid having similar properties (e.g., hydrophobicity, degree and distribution of charged regions), is generally considered in the art to include minor changes. As understood in the art, these minor changes can be identified, in part, by considering the hydropathic index of the amino acid. Kyte et al., J. Mol. Biol. 157: 105-132 (1982). The hydropathic index of an amino acid is based on considerations of its hydrophobicity and charge. It is known in the art that amino acids with similar hydropathic indices can be substituted and still retain protein function. In one aspect, amino acids with a hydropathic index of ± 2 are substituted. The hydrophilicity of amino acids can also be used to reveal substitutions that can produce proteins that retain biological function. Considering the hydrophilicity of amino acids in the context of peptides allows calculation of the maximum local average hydrophilicity of the peptide, which is a useful metric that has been reported to be closely related to antigenicity and immunogenicity. U.S. Patent No. 4,554,101 is fully incorporated herein by reference. As understood in the art, substitutions of amino acids with similar hydrophilicity values ​​can produce peptides that retain biological activity, such as immunogenicity. Amino acids with hydrophilicity values ​​that differ from each other by ± 2 can be substituted. The hydrophobicity index and hydrophilicity value of amino acids are affected by the specific side chains of the amino acids. Consistent with this observation, amino acid substitutions that are compatible with biological functions are considered to depend on the relative similarity of the amino acids, and particularly those amino acid side chains, as revealed by hydrophobicity, hydrophilicity, charge, size, and other properties.

[0087] Variants can be substantially identical nucleotide sequences on the full-length full gene sequence or its fragment. The nucleotide sequence can have 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identity on the full-length gene sequence or its fragment. Variants can be substantially identical amino acid sequences on the full-length amino acid sequence or its fragment. The amino acid sequence can have 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identity on the full-length amino acid sequence or its fragment.

[0088] As used herein, "vector" means a nucleic acid sequence containing an origin of replication. A vector may be a viral vector, a bacteriophage, a bacterial artificial chromosome, or a yeast artificial chromosome. A vector may be a DNA or RNA vector. A vector may be an autonomously replicating extrachromosomal vector, and in one embodiment, an expression plasmid. The vector may contain or include one or more heterologous nucleic acid sequences.

[0089] 2. Synthetic consensus TERT

[0090] The present invention provides an optimized consensus sequence for a TERT antigen. In one embodiment, the antigen encoded by the optimized consensus sequence is capable of eliciting an immune response in a mammal. In one embodiment, the antigen encoded by the optimized consensus sequence may contain one or more epitopes that make it particularly effective as an immunogen that can induce an immune response against it.

[0091] The optimized consensus sequence can be a consensus sequence derived from two or more natural TERT antigens. The optimized consensus sequence may include a consensus sequence and / or one or more modifications for improving expression. Modifications may include codon optimization, RNA optimization, the addition of a kozak sequence to increase translation initiation, and / or the addition of an immunoglobulin leader sequence to enhance immunogenicity. The TERT antigen encoded by the optimized consensus sequence may include a signal peptide, such as an immunoglobulin signal peptide, for example, but not limited to, an immunoglobulin E (IgE) or an immunoglobulin (IgG) signal peptide. In some embodiments, the antigen encoded by the optimized consensus sequence may include a hemagglutinin (HA) tag. The TERT antigen encoded by the optimized consensus sequence can be designed to elicit a stronger cellular and / or humoral immune response than the corresponding natural antigen. The TERT antigen encoded by the optimized consensus sequence can be designed to break tolerance and synergize with anti-cancer immunotherapy.

[0092] In one embodiment, the optimized consensus TERT is designed to break tolerance to native human TERT. In one embodiment, the human optimized consensus TERT coding sequence is as shown in SEQ ID NO: 45, SEQ ID NO: 47, SEQ ID NO: 49, SEQ ID NO: 51, SEQ ID NO: 53, or SEQ ID NO: 55. In one embodiment, the human optimized consensus TERT encoded antigen has an amino acid sequence as shown in SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, SEQ ID NO: 54, or SEQ ID NO: 56.

[0093] In one embodiment, the optimized consensus TERT is designed to break tolerance to native mouse TERT. In one embodiment, the mouse optimized consensus TERT coding sequence is as shown in SEQ ID NO: 49 or SEQ ID NO: 51. In one embodiment, the mouse optimized consensus TERT encoded antigen has the amino acid sequence shown in SEQ ID NO: 50 or SEQ ID NO: 52.

[0094] In one embodiment, the optimized consensus TERT is designed to break tolerance to native rhesus TERT. In one embodiment, the rhesus optimized consensus TERT coding sequence is as shown in SEQ ID NO: 53 or SEQ ID NO: 55. In one embodiment, the rhesus optimized consensus TERT encodes an antigen having an amino acid sequence as shown in SEQ ID NO: 54 or SEQ ID NO: 56.

[0095] In one embodiment, the optimized consensus sequence encoding the TERT antigen is operably linked to one or more regulatory elements. In one embodiment, the regulatory element is a leader sequence. In one embodiment, the optimized consensus DNA sequence operably linked to the IgE leader sequence encoding sequence is shown in SEQ ID NO: 45, SEQ ID NO: 49 and SEQ ID NO: 53. In one embodiment, the optimized consensus sequence encoding the TERT antigen operably linked to the IgE leader sequence is as shown in SEQ ID NO: 46, SEQ ID NO: 50 and SEQ ID NO: 54.

[0096] In one embodiment, the regulatory element is a start codon. Thus, in one embodiment, the present invention relates to a nucleic acid sequence as shown in SEQ ID NO: 47, SEQ ID NO: 51, SEQ ID NO: 55, or a fragment or homolog thereof, operably linked to a nucleotide sequence comprising a start codon at the 5' end. In one embodiment, the present invention relates to an amino acid sequence as shown in SEQ ID NO: 48, SEQ ID NO: 52, or SEQ ID NO: 56, or a fragment or homolog thereof, operably linked to an amino acid encoded by a start codon at the N-terminus (e.g., methionine).

[0097] In one embodiment, the regulatory element is at least one stop codon. Thus, in one embodiment, the present invention relates to the nucleic acid sequence shown in SEQ ID NO:45, SEQ ID NO:47, SEQ ID NO:49, SEQ ID NO:51, SEQ ID NO:53 or SEQ ID NO:55, or a fragment or homologue thereof, operably linked to a nucleotide sequence comprising at least one stop codon at the 3' end. In one embodiment, the nucleotide sequence is operably linked to two stop codons to increase the efficiency of translation termination.

[0098] In one embodiment, the optimized consensus sequence encoding the TERT antigen may encode a peptide having the amino acid sequence shown in SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, SEQ ID NO: 54, or SEQ ID NO: 56. In one embodiment, the optimized consensus sequence may have the nucleotide sequence shown in SEQ ID NO: 45, SEQ ID NO: 47, SEQ ID NO: 49, SEQ ID NO: 51, SEQ ID NO: 53, or SEQ ID NO: 55. In some embodiments, the sequence may be a nucleotide sequence having at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity over the entire length of the nucleotide sequence shown in SEQ ID NO: 45, SEQ ID NO: 47, SEQ ID NO: 49, SEQ ID NO: 51, SEQ ID NO: 53, or SEQ ID NO: 55. In other embodiments, the sequence can be a nucleotide sequence that encodes an amino acid sequence that is at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical over the entire length of the amino acid sequence shown in SEQ ID NO:46, SEQ ID NO:48, SEQ ID NO:50, SEQ ID NO:52, SEQ ID NO:54 or SEQ ID NO:56.

[0099] In some embodiments, the optimized consensus TERT antigen can be encoded by an RNA derived from a DNA sequence that is at least about 96%, 97%, 98%, 99%, or 100% identical over the entire length of the nucleic acid sequence shown in SEQ ID NO: 45, SEQ ID NO: 47, SEQ ID NO: 49, SEQ ID NO: 51, SEQ ID NO: 53, or SEQ ID NO: 55. In some embodiments, the optimized consensus TERT antigen can be encoded by an RNA that encodes an amino acid sequence that is at least about 96%, 97%, 98%, 99%, or 100% identical over the entire length of the amino acid sequence shown in SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, SEQ ID NO: 54, or SEQ ID NO: 56.

[0100] The TERT antigen encoded by the optimized consensus sequence can be a peptide having the amino acid sequence shown in SEQ ID NO: 2, SEQ ID NO: 4, SEQ ID NO: 6, or SEQ ID NO: 8. In some embodiments, the antigen can have an amino acid sequence that is at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical over the entire length of the amino acid sequence shown in SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, SEQ ID NO: 54, or SEQ ID NO: 56.

[0101] Immunogenic fragments of proteins having amino acid sequences homologous to immunogenic fragments of SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, SEQ ID NO: 54, or SEQ ID NO: 56 can be provided. Such immunogenic fragments can comprise at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% of a protein having 95% homology to SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, SEQ ID NO: 54, or SEQ ID NO: 56. Some embodiments relate to immunogenic fragments having 96% homology to immunogenic fragments of the consensus protein sequences herein. Some embodiments relate to immunogenic fragments having 97% homology to immunogenic fragments of the consensus protein sequences herein. Some embodiments relate to immunogenic fragments having 98% homology to an immunogenic fragment of a consensus protein sequence herein. Some embodiments relate to immunogenic fragments having 99% homology to an immunogenic fragment of a consensus protein sequence herein. In some embodiments, the immunogenic fragment comprises a leader sequence, such as, for example, an immunoglobulin leader sequence, such as an IgE leader sequence. In some embodiments, the immunogenic fragment does not contain a leader sequence.

[0102] In one embodiment, the immunogenic fragment of the optimized consensus TERT antigen encodes at least one immunodominant or subimmunodominant epitope of the full-length optimized consensus TERT antigen.

[0103] Some embodiments relate to immunogenic fragments of SEQ ID NO: 45, SEQ ID NO: 47, SEQ ID NO: 49, SEQ ID NO: 51, SEQ ID NO: 53, or SEQ ID NO: 55 that comprise at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% of the full length of SEQ ID NO: 45, SEQ ID NO: 47, SEQ ID NO: 49, SEQ ID NO: 51, SEQ ID NO: 53, or SEQ ID NO: 55. The immunogenic fragment may be at least 96%, at least 97%, at least 98%, or at least 99% homologous to a fragment of SEQ ID NO: 45, SEQ ID NO: 47, SEQ ID NO: 49, SEQ ID NO: 51, SEQ ID NO: 53, or SEQ ID NO: 55. In some embodiments, the immunogenic fragment includes a sequence encoding a leader sequence, such as, for example, an immunoglobulin leader sequence, such as an IgE leader sequence. In some embodiments, the fragment does not contain a coding sequence encoding a leader sequence.

[0104] 3. Immunogenic compositions

[0105] In one embodiment, the immunogenic composition of the present invention may include a synthetic consensus TERT antigen, a fragment thereof, or a variant thereof. In one embodiment, the TERT antigen is a human TERT antigen (hTERT). hTERT is a human telomerase reverse transcriptase that synthesizes a TTAGGG tag at the end of telomerase to prevent cell death caused by chromosome shortening. Hyperproliferative cells may have abnormally high hTERT expression. Abnormal expression of hTERT may also occur in hyperproliferative cells infected with HCV and HPV. Therefore, immunotherapy for HPV and HCV can be enhanced by targeting cells that express hTERT at abnormal levels. HPV and HCV antigens are discussed in more detail below. In one aspect, the hTERT cancer antigen can be further defined by U.S. patent application number 14 / 139,660, filed December 23, 2013, which is incorporated by reference in its entirety.

[0106] In one embodiment, the TERT antigen is a non-human TERT antigen. Non-human TERT antigens include, but are not limited to, mouse TERT (mTERT) and rhesus macaque TERT (rhTERT).

[0107] TERT expression in dendritic cells transfected with TERT gene can induce CD8 + Cytotoxic T cells and induce CD4+ Thus, the use of hTERT expression in antigen presenting cells (APCs) to delay senescence and maintain their ability to present selected antigens can be used in immunotherapies such as those described herein.

[0108] TERT antigens can be associated with or expressed in a variety of cancers, including, but not limited to, melanoma, prostate cancer, liver cancer, cervical cancer, recurrent respiratory papillomatosis (RRP), anal cancer, head and neck cancer, and blood cancers. Thus, when including the TERT antigens described herein, vaccines can be used to treat subjects with a variety of cancers, including, but not limited to, melanoma, prostate cancer, liver cancer, cervical cancer, recurrent respiratory papillomatosis (RRP), anal cancer, head and neck cancer, and blood cancers.

[0109] TERT antigens can induce antigen-specific T cells and / or high-titer antibody responses, thereby inducing or triggering an immune response against a cancer or tumor expressing the antigen or reacting with the cancer or tumor. In some embodiments, the immune response induced or triggered can be a cellular immune response, a humoral immune response, or both a cellular and humoral immune response. In some embodiments, the cellular immune response induced or triggered may include the induction or secretion of interferon-γ (IFN-γ) and / or tumor necrosis factor α (TNF-α). In other embodiments, the immune response induced or triggered can reduce or inhibit one or more immunosuppressive factors that promote the growth of tumors or cancers expressing the antigen, such as, but not limited to, factors that downregulate MHC presentation, factors that upregulate antigen-specific regulatory T cells (Treg), PD-L1, FasL, cytokines such as IL-10 and TFG-β, tumor-associated macrophages, tumor-associated fibroblasts, soluble factors produced by immunosuppressive cells, CTLA-4, PD-1, MDSC, MCP-1, and immune checkpoint molecules, described in more detail below.

[0110] TERT antigens may comprise protein epitopes that make them particularly effective as immunogens against which an anti-TERT immune response can be induced. TERT antigens may comprise full-length translation products, variants thereof, fragments thereof, or combinations thereof. TERT antigens may comprise a consensus protein.

[0111] The nucleic acid sequence encoding TERT antigen or a common TERT antigen can be optimized for codon usage and corresponding RNA transcripts. The nucleic acid encoding TERT antigen or a common TERT antigen can be codons and RNA optimized for expression. In some embodiments, the nucleic acid sequence encoding TERT antigen or a common TERT antigen may include a Kozak sequence (e.g., GCCACC) to enhance the efficiency of translation. The nucleic acid encoding TERT antigen or a common TERT antigen may include multiple stop codons (e.g., TGA TGA) to enhance the efficiency of translation termination.

[0112] The nucleic acid encoding the TERT antigen or the shared TERT antigen may further encode an immunoglobulin E (IgE) leader sequence. The nucleic acid encoding the TERT antigen or the shared TERT antigen may further encode an IgE leader sequence such that the amino acid sequence of the IgE leader sequence is linked to the amino acid sequence of the TERT antigen or the shared TERT antigen by a peptide bond, respectively. The nucleic acid encoding the TERT antigen or the shared TERT antigen may further include a nucleotide sequence encoding the IgE leader sequence. In some embodiments, the nucleic acid encoding the TERT antigen or the shared TERT antigen does not have or contains a nucleotide sequence encoding the IgE leader sequence.

[0113] In some embodiments, the nucleic acid encoding the TERT antigen or the shared TERT antigen can be a heterologous nucleic acid sequence and / or contain one or more heterologous nucleic acid sequences. The nucleic acid encoding the TERT antigen or the shared TERT antigen can be mutated relative to the wild-type TERT antigen so that one or more amino acids or residues in the amino acid sequence of the TERT antigen or the shared TERT antigen are replaced or substituted by another amino acid or residue, respectively. The nucleic acid encoding the TERT antigen or the shared TERT antigen can be mutated relative to the wild-type TERT antigen so that one or more residues in the amino acid sequence of the TERT antigen or the shared TERT antigen are replaced or substituted by another residue, respectively, so that the immune system of a mammal administered with the nucleic acid encoding the TERT antigen or the shared TERT antigen, the TERT antigen or the shared TERT antigen, or a combination thereof is no longer tolerant to TERT. The nucleic acid encoding the TERT antigen or the shared TERT antigen can be mutated relative to the wild-type TERT antigen so that arginine 589, aspartic acid 1005, or arginine 589 and aspartic acid 1005 in the amino acid sequence of the TERT antigen or the shared TERT antigen are replaced or substituted by a tyrosine residue.

[0114] In one aspect, the TERT antigen can be the nucleic acid sequence SEQ ID NO: 23 or SEQ ID NO: 57, which can encode the amino acid sequences SEQ ID NO: 24 and SEQ ID NO: 58, respectively. SEQ ID NO: 23 and SEQ ID NO: 57 encode the TERT antigen linked to an IgE leader sequence. In one embodiment, the TERT antigen can be linked to an IgE leader sequence and an HA tag. In other embodiments, the TERT antigen can be free of or not linked to an IgE leader sequence and / or an HA tag.

[0115] In some embodiments, the TERT antigen can be a nucleic acid sequence that is at least about 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical over the entire length of the nucleic acid sequence shown in SEQ ID NO:23 or SEQ ID NO:57. In other embodiments, the TERT antigen can be a nucleic acid sequence that encodes an amino acid sequence that is at least about 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical over the entire length of the amino acid sequence shown in SEQ ID NO:24 or SEQ ID NO:58. The TERT antigen can be an amino acid sequence that is at least about 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical over the entire length of the amino acid sequence shown in SEQ ID NO:24 or SEQ ID NO:58.

[0116] Some embodiments relate to nucleic acid sequences encoding proteins homologous to the TERT antigen, immunogenic fragments of the TERT antigen, and immunogenic fragments of homologous proteins. Such nucleic acid molecules encoding immunogenic proteins having up to 95% homology to the sequence, up to 96% homology to the sequence, up to 97% homology to the sequence, up to 98% homology to the sequence, and up to 99% homology to the sequence can be provided. Similarly, nucleic acid sequences encoding immunogenic fragments as described herein and immunogenic fragments of proteins homologous to the proteins described herein are also provided.

[0117] Some embodiments relate to nucleic acid molecules encoding immunogenic proteins that have 95% homology to the nucleic acid coding sequences herein. Some embodiments relate to nucleic acid molecules encoding immunogenic proteins that have 96% homology to the nucleic acid coding sequences herein. Some embodiments relate to nucleic acid molecules encoding immunogenic proteins that have 97% homology to the nucleic acid coding sequences herein. Some embodiments relate to nucleic acid molecules encoding immunogenic proteins that have 98% homology to the nucleic acid coding sequences herein. Some embodiments relate to nucleic acid molecules encoding immunogenic proteins that have 99% homology to the nucleic acid coding sequences herein. In some embodiments, the nucleic acid molecules having coding sequences disclosed herein that are homologous to the coding sequences of the consensus proteins disclosed herein include a sequence encoding an IgE leader sequence linked to the 5' end of the coding sequence encoding the homologous protein sequence disclosed herein.

[0118] Some embodiments relate to nucleic acid sequences encoding proteins having a specific percentage of identity to the full-length TERT antigen, immunogenic fragments of the TERT antigen, and immunogenic fragments of proteins having identity to the TERT antigen. Such nucleic acid molecules encoding immunogenic proteins having up to 80% identity to the full-length TERT sequence, up to 85% identity to the full-length sequence, up to 90% identity to the full-length TERT sequence, up to 91% identity to the full-length TERT sequence, up to 92% identity to the full-length TERT sequence, up to 93% identity to the full-length TERT sequence, up to 94% identity to the full-length TERT sequence, up to 95% identity to the full-length TERT sequence, up to 96% identity to the full-length TERT sequence, up to 97% identity to the full-length TERT sequence, up to 98% identity to the full-length TERT sequence, and up to 99% identity to the full-length TERT sequence can be provided. Likewise, nucleic acid sequences encoding immunogenic fragments set forth herein and immunogenic fragments of proteins having similar percent identities to the TERT antigens set forth herein as specified above are also provided.

[0119] In some embodiments, the nucleic acid sequence does not contain a coding sequence that encodes a leader sequence. In some embodiments, the nucleic acid sequence does not contain a coding sequence that encodes an IgE leader sequence.

[0120] Some embodiments relate to fragments of SEQ ID NO: 23 or SEQ ID NO: 57. The fragments can be at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% of SEQ ID NO: 23 or SEQ ID NO: 57. The fragments can be at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% homologous to a fragment of SEQ ID NO: 23 or SEQ ID NO: 57. The fragment can be at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to a fragment of SEQ ID NO: 23 or SEQ ID NO: 57. In some embodiments, the fragment includes a sequence encoding a leader sequence, such as, for example, an immunoglobulin leader sequence, such as an IgE leader sequence. In some embodiments, the fragment does not contain a coding sequence encoding a leader sequence. In some embodiments, the fragment does not contain a coding sequence encoding a leader sequence, such as, for example, an IgE leader sequence.

[0121] In addition, the amino acid sequence of the TERT antigen is SEQ ID NO: 24 or SEQ ID NO: 58. The amino acid sequence of the TERT antigen linked to the IgE leader sequence is SEQ ID NO: 24 or SEQ ID NO: 58. The amino acid sequence of the TERT antigen linked to the IgE leader sequence may be linked to an HA tag.

[0122] Some embodiments relate to proteins that are homologous to SEQ ID NO: 24 or SEQ ID NO: 58. Some embodiments relate to immunogenic proteins that are 95% homologous to the protein sequence shown in SEQ ID NO: 24 or SEQ ID NO: 58. Some embodiments relate to immunogenic proteins that are 96% homologous to the protein sequence shown in SEQ ID NO: 24 or SEQ ID NO: 58. Some embodiments relate to immunogenic proteins that are 97% homologous to the protein sequence shown in SEQ ID NO: 24 or SEQ ID NO: 58. Some embodiments relate to immunogenic proteins that are 98% homologous to the protein sequence shown in SEQ ID NO: 24 or SEQ ID NO: 58. Some embodiments relate to immunogenic proteins that are 99% homologous to the protein sequence shown in SEQ ID NO: 24 or SEQ ID NO: 58.

[0123] Some embodiments relate to proteins identical to SEQ ID NO: 24 or SEQ ID NO: 58. Some embodiments relate to immunogenic proteins having an amino acid sequence that is 80% identical to the full-length amino acid sequence shown in SEQ ID NO: 24 or SEQ ID NO: 58. Some embodiments relate to immunogenic proteins having an amino acid sequence that is 85% identical to the full-length amino acid sequence shown in SEQ ID NO: 24 or SEQ ID NO: 58. Some embodiments relate to immunogenic proteins having an amino acid sequence that is 90% identical to the full-length amino acid sequence shown in SEQ ID NO: 24 or SEQ ID NO: 58. Some embodiments relate to immunogenic proteins having an amino acid sequence that is 91% identical to the full-length amino acid sequence shown in SEQ ID NO: 24 or SEQ ID NO: 58. Some embodiments relate to immunogenic proteins having an amino acid sequence that is 92% identical to the full-length amino acid sequence shown in SEQ ID NO: 24 or SEQ ID NO: 58. Some embodiments relate to immunogenic proteins having an amino acid sequence that is 93% identical to the full-length amino acid sequence shown in SEQ ID NO: 24 or SEQ ID NO: 58. Some embodiments relate to immunogenic proteins having an amino acid sequence that is 94% identical to the full-length amino acid sequence shown in SEQ ID NO: 24 or SEQ ID NO: 58. Some embodiments relate to immunogenic proteins having an amino acid sequence that is 95% identical to the full-length amino acid sequence shown in SEQ ID NO: 24 or SEQ ID NO: 58. Some embodiments relate to immunogenic proteins having an amino acid sequence that is 96% identical to the full-length amino acid sequence shown in SEQ ID NO: 24 or SEQ ID NO: 58. Some embodiments relate to immunogenic proteins having an amino acid sequence that is 97% identical to the full-length amino acid sequence shown in SEQ ID NO: 24 or SEQ ID NO: 58. Some embodiments relate to immunogenic proteins having an amino acid sequence that is 98% identical to the full-length amino acid sequence shown in SEQ ID NO:24 or SEQ ID NO:58. Some embodiments relate to immunogenic proteins having an amino acid sequence that is 99% identical to the full-length amino acid sequence shown in SEQ ID NO:24 or SEQ ID NO:58.

[0124] In some embodiments, the protein does not contain a leader sequence. In some embodiments, the protein does not contain an IgE leader sequence. A fragment of a protein may comprise at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% of the protein. An immunogenic fragment of SEQ ID NO: 24 or SEQ ID NO: 58 may be provided. The immunogenic fragment may comprise at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% of SEQ ID NO: 24 or SEQ ID NO: 58. In some embodiments, the fragment includes a leader sequence, such as, for example, an immunoglobulin leader sequence, such as an IgE leader sequence. In some embodiments, the fragment does not contain a leader sequence. In some embodiments, the fragment does not contain a leader sequence, such as, for example, an IgE leader sequence.

[0125] Immunogenic fragments of proteins having amino acid sequences homologous to immunogenic fragments of SEQ ID NO: 24 or SEQ ID NO: 58 can be provided. Such immunogenic fragments can comprise at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% of a protein having 95% or greater homology to SEQ ID NO: 18. Some embodiments relate to immunogenic fragments having 96% homology to immunogenic fragments of the protein sequences herein. Some embodiments relate to immunogenic fragments having 97% homology to immunogenic fragments of the protein sequences herein. Some embodiments relate to immunogenic fragments having 98% homology to an immunogenic fragment of a protein sequence herein. Some embodiments relate to immunogenic fragments having 99% homology to an immunogenic fragment of a protein sequence herein. In some embodiments, the fragment includes a leader sequence, such as, for example, an immunoglobulin leader sequence, such as an IgE leader sequence. In some embodiments, the fragment does not contain a leader sequence. In some embodiments, the fragment does not contain a leader sequence, such as, for example, an IgE leader sequence.

[0126] Immunogenic fragments of a protein can be provided that have an amino acid sequence identical to an immunogenic fragment of SEQ ID NO: 24 or SEQ ID NO: 58. Such immunogenic fragments can comprise at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% of a protein having 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to the amino acid sequence shown in SEQ ID NO: 24 or SEQ ID NO: 58. In some embodiments, the fragment includes a leader sequence, such as, for example, an immunoglobulin leader sequence, such as an IgE leader sequence. In some embodiments, the fragment does not contain a leader sequence. In some embodiments, the fragment does not contain a leader sequence, such as, for example, an IgE leader sequence.

[0127] As indicated herein with respect to linking a signal peptide or leader sequence to the N-terminus of a protein, the signal peptide / leader sequence replaces the protein's N-terminal methionine that is encoded by the start codon of the nucleic acid sequence encoding the protein without the signal peptide coding sequence.

[0128] A fragment of SEQ ID NO: 23 or SEQ ID NO: 57 may comprise 30 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 23 or SEQ ID NO: 57 may comprise 45 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 23 or SEQ ID NO: 57 may comprise 60 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 23 or SEQ ID NO: 57 may comprise 75 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 23 or SEQ ID NO: 57 may comprise 90 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 23 or SEQ ID NO: 57 may comprise 120 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 23 or SEQ ID NO: 57 may comprise 150 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 23 or SEQ ID NO: 57 may comprise 180 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 23 or SEQ ID NO: 57 may comprise 210 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 23 or SEQ ID NO: 57 may comprise 240 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 23 or SEQ ID NO: 57 may comprise 270 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 23 or SEQ ID NO: 57 may comprise 300 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 23 or SEQ ID NO: 57 may comprise 360 ​​or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 23 or SEQ ID NO: 57 may comprise 420 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 23 or SEQ ID NO: 57 may comprise 480 or more nucleotides, including a sequence encoding an immunodominant epitope.In some embodiments, a fragment of SEQ ID NO: 23 or SEQ ID NO: 57 may comprise 540 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 23 or SEQ ID NO: 57 may comprise 600 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 23 or SEQ ID NO: 57 may comprise 300 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 23 or SEQ ID NO: 57 may comprise 660 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 23 or SEQ ID NO: 57 may comprise 720 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 23 or SEQ ID NO: 57 may comprise 780 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 23 or SEQ ID NO: 57 may comprise 840 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 23 or SEQ ID NO: 57 may comprise 900 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 23 or SEQ ID NO: 57 may comprise 960 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 23 or SEQ ID NO: 57 may comprise 1020 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 23 or SEQ ID NO: 57 may comprise 1080 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 23 or SEQ ID NO: 57 may comprise 1140 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 23 or SEQ ID NO: 57 may comprise 1200 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 23 or SEQ ID NO: 57 may comprise 1260 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 23 or SEQ ID NO: 57 may comprise 1320 or more nucleotides, including a sequence encoding an immunodominant epitope.In some embodiments, a fragment of SEQ ID NO: 23 or SEQ ID NO: 57 may comprise 1380 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 23 or SEQ ID NO: 57 may comprise 1440 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 23 or SEQ ID NO: 57 may comprise 1500 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 23 or SEQ ID NO: 57 may comprise 1560 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 23 or SEQ ID NO: 57 may comprise 1620 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 23 or SEQ ID NO: 57 may comprise 1680 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 23 or SEQ ID NO: 57 may comprise 1740 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 23 or SEQ ID NO: 57 may comprise 1800 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 23 or SEQ ID NO: 57 may comprise 1860 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 23 or SEQ ID NO: 57 may comprise 1920 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 23 or SEQ ID NO: 57 may comprise 1980 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 23 or SEQ ID NO: 57 may comprise 2040 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 23 or SEQ ID NO: 57 may comprise 2100 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 23 or SEQ ID NO: 57 may comprise 2160 or more nucleotides, including a sequence encoding an immunodominant epitope.In some embodiments, a fragment of SEQ ID NO: 23 or SEQ ID NO: 57 may comprise 2220 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 23 or SEQ ID NO: 57 may comprise 2280 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 23 or SEQ ID NO: 57 may comprise 2340 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 23 or SEQ ID NO: 57 may comprise 2400 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 23 or SEQ ID NO: 57 may comprise 2460 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 23 or SEQ ID NO: 57 may comprise 2520 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 23 or SEQ ID NO: 57 may comprise 2580 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 23 or SEQ ID NO: 57 may comprise 2640 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 23 or SEQ ID NO: 57 may comprise 2700 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 23 or SEQ ID NO: 57 may comprise 2760 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 23 or SEQ ID NO: 57 may comprise 2820 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 23 or SEQ ID NO: 57 may comprise 2880 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 23 or SEQ ID NO: 57 may comprise 2940 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 23 or SEQ ID NO: 57 may comprise 3000 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 23 or SEQ ID NO: 57 may comprise 3060 or more nucleotides, including a sequence encoding an immunodominant epitope.In some embodiments, a fragment of SEQ ID NO: 23 or SEQ ID NO: 57 may comprise 3120 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 23 or SEQ ID NO: 57 may comprise 3180 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 23 or SEQ ID NO: 57 may comprise 3240 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 23 or SEQ ID NO: 57 may comprise 3300 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 23 or SEQ ID NO: 57 may comprise 3360 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 23 or SEQ ID NO: 57 may comprise 3420 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 23 or SEQ ID NO: 57 may comprise 3480 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 23 or SEQ ID NO: 57 may comprise a sequence encoding an IgE leader sequence. In some embodiments, a fragment of SEQ ID NO: 23 or SEQ ID NO: 57 does not comprise a sequence encoding an IgE leader sequence.

[0129] A fragment may comprise less than 60 nucleotides, in some embodiments less than 75 nucleotides, in some embodiments less than 90 nucleotides, in some embodiments less than 120 nucleotides, in some embodiments less than 150 nucleotides, in some embodiments less than 180 nucleotides, in some embodiments less than 210 nucleotides, in some embodiments less than 240 nucleotides, in some embodiments less than 270 nucleotides, in some embodiments less than 300 nucleotides, in some embodiments less than 360 nucleotides, in some embodiments less than 420 nucleotides, in some embodiments less than 480 nucleotides, in some embodiments less than 540 nucleotides, in some embodiments less than 600 nucleotides. In some embodiments, less than 600 nucleotides, in some embodiments less than 660 nucleotides, in some embodiments less than 720 nucleotides, in some embodiments less than 780 nucleotides, in some embodiments less than 840 nucleotides, in some embodiments less than 900 nucleotides, in some embodiments less than 960 nucleotides, in some embodiments less than 1020 nucleotides, in some embodiments less than 1080 nucleotides, in some embodiments less than 1140 nucleotides, in some embodiments less than 1200 nucleotides, in some embodiments less than 1260 nucleotides, in some embodiments less than 1320 nucleotides, in some embodiments less than 1380 nucleotides. 0 nucleotides, in some embodiments less than 1440 nucleotides, in some embodiments less than 1500 nucleotides, in some embodiments less than 1560 nucleotides, in some embodiments less than 1620 nucleotides, in some embodiments less than 1680 nucleotides, in some embodiments less than 1740 nucleotides, in some embodiments less than 1800 nucleotides, in some embodiments less than 1860 nucleotides, in some embodiments less than 1920 nucleotides, in some embodiments less than 1980 nucleotides, in some embodiments less than 2040 nucleotides, in some embodiments less than 2100 nucleotides, in some embodiments less than 2160 nucleotides nucleotides, in some embodiments less than 2220 nucleotides, in some embodiments less than 2280 nucleotides, in some embodiments less than 2340 nucleotides, in some embodiments less than 2400 nucleotides, in some embodiments less than 2460 nucleotides, in some embodiments less than 2520 nucleotides, in some embodiments less than 2580 nucleotides, in some embodiments less than 2640 nucleotides, in some embodiments less than 2700 nucleotides, in some embodiments less than 2760 nucleotides, in some embodiments less than 2820 nucleotides, in some embodiments less than 2860 nucleotides, in some embodiments less than 2940 nucleotides,In some embodiments, less than 3000 nucleotides, in some embodiments, less than 3060 nucleotides, in some embodiments, less than 3120 nucleotides, in some embodiments, less than 3180 nucleotides, in some embodiments, less than 3240 nucleotides, in some embodiments, less than 3300 nucleotides, in some embodiments, less than 3360 nucleotides, in some embodiments, less than 3420 nucleotides, in some embodiments, less than 3480 nucleotides, and in some embodiments, less than 3510 nucleotides.

[0130] Fragments of SEQ ID NO: 24 or SEQ ID NO: 58 may comprise 15 or more amino acids, including sequences encoding immunodominant epitopes. In some embodiments, fragments of SEQ ID NO: 24 or SEQ ID NO: 58 may comprise 18 or more amino acids, including sequences encoding immunodominant epitopes. In some embodiments, fragments of SEQ ID NO: 24 or SEQ ID NO: 58 may comprise 21 or more amino acids, including sequences encoding immunodominant epitopes. In some embodiments, fragments of SEQ ID NO: 24 or SEQ ID NO: 58 may comprise 24 or more amino acids, including sequences encoding immunodominant epitopes. In some embodiments, fragments of SEQ ID NO: 24 or SEQ ID NO: 58 may comprise 30 or more amino acids, including sequences encoding immunodominant epitopes. In some embodiments, fragments of SEQ ID NO: 24 or SEQ ID NO: 58 may comprise 36 or more amino acids, including sequences encoding immunodominant epitopes. In some embodiments, a fragment of SEQ ID NO: 24 or SEQ ID NO: 58 may comprise 42 or more amino acids, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 24 or SEQ ID NO: 58 may comprise 48 or more amino acids, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 24 or SEQ ID NO: 58 may comprise 54 or more amino acids, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 24 or SEQ ID NO: 58 may comprise 60 or more amino acids, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 24 or SEQ ID NO: 58 may comprise 66 or more amino acids, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 24 or SEQ ID NO: 58 may comprise 72 or more amino acids, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 24 or SEQ ID NO: 58 may comprise 90 or more amino acids, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 24 or SEQ ID NO: 58 may comprise 120 or more amino acids, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 24 or SEQ ID NO: 58 may comprise 150 or more amino acids, including a sequence encoding an immunodominant epitope.In some embodiments, a fragment of SEQ ID NO: 24 or SEQ ID NO: 58 may comprise 180 or more amino acids, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 24 or SEQ ID NO: 58 may comprise 210 or more amino acids, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 24 or SEQ ID NO: 58 may comprise 240 or more amino acids, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 24 or SEQ ID NO: 58 may comprise 270 or more amino acids, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 24 or SEQ ID NO: 58 may comprise 300 or more amino acids, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 24 or SEQ ID NO: 58 may comprise 330 or more amino acids, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 24 or SEQ ID NO: 58 may comprise 360 ​​or more amino acids, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 24 or SEQ ID NO: 58 may comprise 390 or more amino acids, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 24 or SEQ ID NO: 58 may comprise 420 or more amino acids, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 24 or SEQ ID NO: 58 may comprise 450 or more amino acids, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 24 or SEQ ID NO: 58 may comprise 480 or more amino acids, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 24 or SEQ ID NO: 58 may comprise 510 or more amino acids, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 24 or SEQ ID NO: 58 may comprise 540 or more amino acids, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 24 or SEQ ID NO: 58 may comprise 570 or more amino acids, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 24 or SEQ ID NO: 58 may comprise 600 or more amino acids, including a sequence encoding an immunodominant epitope.In some embodiments, a fragment of SEQ ID NO: 24 or SEQ ID NO: 58 may comprise 630 or more amino acids, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 24 or SEQ ID NO: 58 may comprise 660 or more amino acids, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 24 or SEQ ID NO: 58 may comprise 690 or more amino acids, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 24 or SEQ ID NO: 58 may comprise 720 or more amino acids, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 24 or SEQ ID NO: 58 may comprise 750 or more amino acids, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 24 or SEQ ID NO: 58 may comprise 780 or more amino acids, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 24 or SEQ ID NO: 58 may comprise 810 or more amino acids, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 24 or SEQ ID NO: 58 may comprise 840 or more amino acids, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 24 or SEQ ID NO: 58 may comprise 870 or more amino acids, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 24 or SEQ ID NO: 58 may comprise 900 or more amino acids, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 24 or SEQ ID NO: 58 may comprise 930 or more amino acids, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 24 or SEQ ID NO: 58 may comprise 960 or more amino acids, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 24 or SEQ ID NO: 58 may comprise 990 or more amino acids, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 24 or SEQ ID NO: 58 may comprise 1020 or more amino acids, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 24 or SEQ ID NO: 58 may comprise 1050 or more amino acids, including a sequence encoding an immunodominant epitope.In some embodiments, a fragment of SEQ ID NO: 24 or SEQ ID NO: 58 may comprise 1080 or more amino acids, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 24 or SEQ ID NO: 58 may comprise 1110 or more amino acids, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 24 or SEQ ID NO: 58 may comprise 1140 or more amino acids, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 24 or SEQ ID NO: 58 may comprise 1170 or more amino acids, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 24 or SEQ ID NO: 58 may comprise 1200 or more amino acids, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 24 or SEQ ID NO: 58 may comprise 1230 or more amino acids, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 24 or SEQ ID NO: 58 may comprise 1260 or more amino acids, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 24 or SEQ ID NO: 58 may comprise 1290 or more amino acids, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 24 or SEQ ID NO: 58 may comprise 1320 or more amino acids, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 24 or SEQ ID NO: 58 may comprise 1350 or more amino acids, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 24 or SEQ ID NO: 58 may comprise 1380 or more amino acids, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 24 or SEQ ID NO: 58 may comprise 1410 or more amino acids, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 24 or SEQ ID NO: 58 may comprise 1440 or more amino acids, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 24 or SEQ ID NO: 58 may comprise 1470 or more amino acids, including a sequence encoding an immunodominant epitope.In some embodiments, a fragment of SEQ ID NO: 24 or SEQ ID NO: 58 may comprise 1500 or more amino acids, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 24 or SEQ ID NO: 58 may comprise a coding sequence for an IgE leader sequence. In some embodiments, a fragment of SEQ ID NO: 24 or SEQ ID NO: 58 does not comprise a coding sequence for an IgE leader sequence.

[0131] Fragments may comprise less than 24 amino acids, in some embodiments less than 30 amino acids, in some embodiments less than 36 amino acids, in some embodiments less than 42 amino acids, in some embodiments less than 48 amino acids, in some embodiments less than 54 amino acids, in some embodiments less than 60 amino acids, in some embodiments less than 72 amino acids, in some embodiments less than 90 amino acids, in some embodiments less than 120 amino acids, in some embodiments less than 150 amino acids, in some embodiments less than 180 amino acids, in some embodiments less than 210 amino acids, in some embodiments less than 240 amino acids, in some embodiments less than 260 amino acids, in some embodiments less than 290 amino acids, in some embodiments less than 320 amino acids, in some embodiments less than 350 amino acids, in some embodiments less than 380 amino acids, in some embodiments less than 410 amino acids, in some embodiments less than 440 amino acids, in some embodiments less than 470 amino acids, in some embodiments less than 500 amino acids, in some embodiments less than 530 amino acids, in some embodiments less than 560 amino acids, in some embodiments less than 590 amino acids, in some embodiments less than 620 amino acids, in some embodiments less than 640 amino acids. In some embodiments, the protein comprises less than 650 amino acids, in some embodiments less than 680 amino acids, in some embodiments less than 710 amino acids, in some embodiments less than 740 amino acids, in some embodiments less than 770 amino acids, in some embodiments less than 800 amino acids, in some embodiments less than 830 amino acids, in some embodiments less than 860 amino acids, in some embodiments less than 890 amino acids, in some embodiments less than 920 amino acids, in some embodiments less than 950 amino acids, in some embodiments less than 980 amino acids, in some embodiments less than 1010 amino acids, in some embodiments less than In some embodiments, the present invention comprises less than 1040 amino acids, in some embodiments less than 1070 amino acids, in some embodiments less than 1200 amino acids, in some embodiments less than 1230 amino acids, in some embodiments less than 1260 amino acids, in some embodiments less than 1290 amino acids, in some embodiments less than 1320 amino acids, in some embodiments less than 1350 amino acids, in some embodiments less than 1380 amino acids, in some embodiments less than 1410 amino acids, in some embodiments less than 1440 amino acids, in some embodiments less than 1470 amino acids, and in some embodiments less than 1500 amino acids.

[0132] In one embodiment, the TERT antigen is a synthetic consensus TERT antigen. In certain embodiments, the synthetic consensus TERT comprises 1 or more, 2 or more, 3 or more, 4 or more, 5 or more, 6 or more, 7 or more, 8 or more, 9 or more, 10 or more, 15 or more, 20 or more, 30 or more, or 50 or more amino acid mutations relative to the native TERT antigen.

[0133] In one embodiment, the consensus hTERT antigen can be the nucleic acid sequence of SEQ ID NO:45, which encodes the amino acid sequence of SEQ ID NO:46. SEQ ID NO:45 encodes the consensus hTERT antigen linked to an IgE leader sequence. The consensus hTERT antigen can be linked to an IgE leader sequence and an HA tag. In other embodiments, the consensus hTERT antigen can be free of or not linked to an IgE leader sequence and / or an HA tag. In one embodiment, the consensus hTERT antigen can be the nucleic acid sequence of SEQ ID NO:47, which encodes the amino acid sequence of SEQ ID NO:48. SEQ ID NO:47 encodes the consensus hTERT antigen free of or not linked to an IgE leader sequence and / or an HA tag.

[0134] In one embodiment, the consensus mTERT antigen can be the nucleic acid sequence SEQ ID NO: 49, which encodes the amino acid sequence SEQ ID NO: 50. SEQ ID NO: 49 encodes the consensus mTERT antigen linked to an IgE leader sequence. The consensus mTERT antigen can be linked to an IgE leader sequence and an HA tag. In other embodiments, the consensus mTERT antigen can be free of or not linked to an IgE leader sequence and / or an HA tag. In one embodiment, the consensus mTERT antigen can be the nucleic acid sequence SEQ ID NO: 51, which encodes the amino acid sequence SEQ ID NO: 52. SEQ ID NO: 51 encodes the consensus mTERT antigen free of or not linked to an IgE leader sequence and / or an HA tag.

[0135] In one embodiment, the consensus rhTERT antigen can be the nucleic acid sequence of SEQ ID NO: 53, which encodes the amino acid sequence of SEQ ID NO: 54. SEQ ID NO: 53 encodes the consensus rhTERT antigen linked to an IgE leader sequence. The consensus rhTERT antigen can be linked to an IgE leader sequence and an HA tag. In other embodiments, the consensus rhTERT antigen can be free of or not linked to an IgE leader sequence and / or an HA tag. In one embodiment, the consensus rhTERT antigen can be the nucleic acid sequence of SEQ ID NO: 55, which encodes the amino acid sequence of SEQ ID NO: 56. SEQ ID NO: 55 encodes the consensus rhTERT antigen free of or not linked to an IgE leader sequence and / or an HA tag.

[0136] In some embodiments, the consensus TERT antigen can be a nucleic acid sequence that is at least about 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical over the entire length of the nucleic acid sequence shown in SEQ ID NO:45, SEQ ID NO:47, SEQ ID NO:49, SEQ ID NO:51, SEQ ID NO:53, or SEQ ID NO:55. In other embodiments, the consensus TERT antigen can be a nucleic acid sequence that encodes an amino acid sequence that is at least about 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical over the entire length of the amino acid sequence shown in SEQ ID NO:46, SEQ ID NO:48, SEQ ID NO:50, SEQ ID NO:52, SEQ ID NO:54 or SEQ ID NO:56. The consensus TERT antigen can be an amino acid sequence that is at least about 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical over the entire length of the amino acid sequence shown in SEQ ID NO:46, SEQ ID NO:48, SEQ ID NO:50, SEQ ID NO:52, SEQ ID NO:54 or SEQ ID NO:56.

[0137] Some embodiments relate to nucleic acid sequences encoding proteins homologous to the consensus TERT antigen, immunogenic fragments of the consensus TERT antigen, and immunogenic fragments of homologous proteins. Such nucleic acid molecules encoding immunogenic proteins having up to 95% homology to the sequence, up to 96% homology to the sequence, up to 97% homology to the sequence, up to 98% homology to the sequence, and up to 99% homology to the sequence can be provided. Similarly, nucleic acid sequences encoding immunogenic fragments as set forth herein and immunogenic fragments of proteins homologous to the proteins set forth herein are also provided.

[0138] Some embodiments relate to nucleic acid molecules encoding immunogenic proteins that have 95% homology to the nucleic acid coding sequences herein. Some embodiments relate to nucleic acid molecules encoding immunogenic proteins that have 96% homology to the nucleic acid coding sequences herein. Some embodiments relate to nucleic acid molecules encoding immunogenic proteins that have 97% homology to the nucleic acid coding sequences herein. Some embodiments relate to nucleic acid molecules encoding immunogenic proteins that have 98% homology to the nucleic acid coding sequences herein. Some embodiments relate to nucleic acid molecules encoding immunogenic proteins that have 99% homology to the nucleic acid coding sequences herein. In some embodiments, the nucleic acid molecules having coding sequences disclosed herein that are homologous to the coding sequences of the consensus proteins disclosed herein include a sequence encoding an IgE leader sequence linked to the 5' end of the coding sequence encoding the homologous protein sequence disclosed herein.

[0139] Some embodiments relate to nucleic acid sequences encoding proteins having a specific percentage identity to the full-length consensus TERT antigen, immunogenic fragments of the consensus TERT antigen, and immunogenic fragments of proteins having identity to the consensus TERT antigen. Such nucleic acid molecules can be provided that encode immunogenic proteins having up to 80% identity to the full-length consensus TERT sequence, up to 85% identity to the full-length consensus sequence, up to 90% identity to the full-length consensus TERT sequence, up to 91% identity to the full-length consensus TERT sequence, up to 92% identity to the full-length consensus TERT sequence, up to 93% identity to the full-length consensus TERT sequence, up to 94% identity to the full-length consensus TERT sequence, up to 95% identity to the full-length consensus TERT sequence, up to 96% identity to the full-length consensus TERT sequence, up to 97% identity to the full-length consensus TERT sequence, up to 98% identity to the full-length consensus TERT sequence, up to 99% identity to the full-length consensus TERT sequence. Similarly, nucleic acid sequences encoding immunogenic fragments set forth herein and immunogenic fragments of proteins having similar percentages of identity as specified above to the consensus TERT antigens set forth herein are also provided.

[0140] In some embodiments, the nucleic acid sequence does not contain a coding sequence that encodes a leader sequence. In some embodiments, the nucleic acid sequence does not contain a coding sequence that encodes an IgE leader sequence.

[0141] Some embodiments relate to fragments of SEQ ID NO: 45, SEQ ID NO: 47, SEQ ID NO: 49, SEQ ID NO: 51, SEQ ID NO: 53, or SEQ ID NO: 55. The fragment can be at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% of SEQ ID NO: 45, SEQ ID NO: 47, SEQ ID NO: 49, SEQ ID NO: 51, SEQ ID NO: 53, or SEQ ID NO: 55. Fragments can be at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% homologous to a fragment of SEQ ID NO: 45, SEQ ID NO: 47, SEQ ID NO: 49, SEQ ID NO: 51, SEQ ID NO: 53, or SEQ ID NO: 55. Fragments can be at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to a fragment of SEQ ID NO: 45, SEQ ID NO: 47, SEQ ID NO: 49, SEQ ID NO: 51, SEQ ID NO: 53, or SEQ ID NO: 55. In some embodiments, the fragments include a sequence encoding a leader sequence, such as, for example, an immunoglobulin leader sequence, such as an IgE leader sequence. In some embodiments, the fragments do not contain a coding sequence encoding a leader sequence. In some embodiments, the fragment does not contain coding sequences encoding a leader sequence, such as, for example, an IgE leader sequence.

[0142] In addition, in one embodiment, the amino acid sequence of the consensus TERT antigen is SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, SEQ ID NO: 54, or SEQ ID NO: 56. The amino acid sequence of the consensus TERT antigen linked to the IgE leader sequence is SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, SEQ ID NO: 54, or SEQ ID NO: 56. The amino acid sequence of the consensus TERT antigen linked to the IgE leader sequence can be linked to an HA tag.

[0143] Some embodiments relate to proteins homologous to SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, SEQ ID NO: 54, or SEQ ID NO: 56. Some embodiments relate to immunogenic proteins that are 95% homologous to the protein sequence shown in SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, SEQ ID NO: 54, or SEQ ID NO: 56. Some embodiments relate to immunogenic proteins that are 96% homologous to the protein sequence shown in SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, SEQ ID NO: 54, or SEQ ID NO: 56. Some embodiments relate to immunogenic proteins that are 97% homologous to the protein sequence shown in SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, SEQ ID NO: 54, or SEQ ID NO: 56. Some embodiments relate to immunogenic proteins that are 98% homologous to the protein sequence shown in SEQ ID NO: 46. Some embodiments relate to immunogenic proteins that are 99% homologous to the protein sequence shown in SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, SEQ ID NO: 54, or SEQ ID NO: 56.

[0144] Some embodiments relate to proteins identical to SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, SEQ ID NO: 54, or SEQ ID NO: 56. Some embodiments relate to immunogenic proteins having an amino acid sequence that is 80% identical to the full-length amino acid sequence shown in SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, SEQ ID NO: 54, or SEQ ID NO: 56. Some embodiments relate to immunogenic proteins having an amino acid sequence that is 85% identical to the full-length amino acid sequence shown in SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, SEQ ID NO: 54, or SEQ ID NO: 56. Some embodiments relate to immunogenic proteins having an amino acid sequence that is 90% identical to the full-length amino acid sequence shown in SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, SEQ ID NO: 54, or SEQ ID NO: 56. Some embodiments relate to immunogenic proteins having an amino acid sequence that is 91% identical to the full-length amino acid sequence shown in SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, SEQ ID NO: 54, or SEQ ID NO: 56. Some embodiments relate to immunogenic proteins having an amino acid sequence that is 92% identical to the full-length amino acid sequence shown in SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, SEQ ID NO: 54, or SEQ ID NO: 56. Some embodiments relate to immunogenic proteins having an amino acid sequence that is 93% identical to the full-length amino acid sequence shown in SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, SEQ ID NO: 54, or SEQ ID NO: 56. Some embodiments relate to immunogenic proteins having an amino acid sequence that is 94% identical to the full-length amino acid sequence shown in SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, SEQ ID NO: 54, or SEQ ID NO: 56.Some embodiments relate to immunogenic proteins having an amino acid sequence that is 95% identical to the full-length amino acid sequence shown in SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, SEQ ID NO: 54, or SEQ ID NO: 56. Some embodiments relate to immunogenic proteins having an amino acid sequence that is 96% identical to the full-length amino acid sequence shown in SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, SEQ ID NO: 54, or SEQ ID NO: 56. Some embodiments relate to immunogenic proteins having an amino acid sequence that is 97% identical to the full-length amino acid sequence shown in SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, SEQ ID NO: 54, or SEQ ID NO: 56. Some embodiments relate to immunogenic proteins having an amino acid sequence that is 98% identical to the full-length amino acid sequence shown in SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, SEQ ID NO: 54, or SEQ ID NO: 56. Some embodiments relate to immunogenic proteins having an amino acid sequence that is 99% identical to the full-length amino acid sequence shown in SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, SEQ ID NO: 54, or SEQ ID NO: 56.

[0145] In some embodiments, the protein does not contain a leader sequence. In some embodiments, the protein does not contain an IgE leader sequence. A fragment of a protein can comprise at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% of the protein. Immunogenic fragments of SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, SEQ ID NO: 54, or SEQ ID NO: 56 can be provided. The immunogenic fragment may comprise at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% of SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, SEQ ID NO: 54, or SEQ ID NO: 56. In some embodiments, the fragment includes a leader sequence, such as, for example, an immunoglobulin leader sequence, such as an IgE leader sequence. In some embodiments, the fragment does not contain a leader sequence. In some embodiments, the fragment does not contain a leader sequence, such as, for example, an IgE leader sequence.

[0146] Immunogenic fragments of proteins having amino acid sequences homologous to immunogenic fragments of SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, SEQ ID NO: 54, or SEQ ID NO: 56 can be provided. Such immunogenic fragments can comprise at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% of a protein having 95% or greater homology to SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, SEQ ID NO: 54, or SEQ ID NO: 56. Some embodiments relate to immunogenic fragments with 96% homology to the immunogenic fragments of the protein sequences herein. Some embodiments relate to immunogenic fragments with 97% homology to the immunogenic fragments of the protein sequences herein. Some embodiments relate to immunogenic fragments with 98% homology to the immunogenic fragments of the protein sequences herein. Some embodiments relate to immunogenic fragments with 99% homology to the immunogenic fragments of the protein sequences herein. In some embodiments, the fragments include a leader sequence, such as, for example, an immunoglobulin leader sequence, such as an IgE leader sequence. In some embodiments, the fragments do not contain a leader sequence. In some embodiments, the fragments do not contain a leader sequence, such as, for example, an IgE leader sequence.

[0147] Immunogenic fragments of the protein can be provided that have the same amino acids as an immunogenic fragment of SEQ ID NO:46, SEQ ID NO:48, SEQ ID NO:50, SEQ ID NO:52, SEQ ID NO:54, or SEQ ID NO:56. Such immunogenic fragments may comprise at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% of a protein having 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to the amino acid sequence shown in SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, SEQ ID NO: 54, or SEQ ID NO: 56. In some embodiments, the fragment includes a leader sequence, such as, for example, an immunoglobulin leader sequence, such as an IgE leader sequence. In some embodiments, the fragments do not contain a leader sequence.In some embodiments, the fragments do not contain a leader sequence, such as, for example, an IgE leader sequence.

[0148] As indicated herein with respect to linking a signal peptide or leader sequence to the N-terminus of a protein, the signal peptide / leader sequence replaces the protein's N-terminal methionine that is encoded by the start codon of the nucleic acid sequence encoding the protein without the signal peptide coding sequence.

[0149] A fragment of SEQ ID NO: 45, SEQ ID NO: 47, SEQ ID NO: 49, SEQ ID NO: 51, SEQ ID NO: 53, or SEQ ID NO: 55 may comprise 30 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 45, SEQ ID NO: 47, SEQ ID NO: 49, SEQ ID NO: 51, SEQ ID NO: 53, or SEQ ID NO: 55 may comprise 45 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 45, SEQ ID NO: 47, SEQ ID NO: 49, SEQ ID NO: 51, SEQ ID NO: 53, or SEQ ID NO: 55 may comprise 60 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 45, SEQ ID NO: 47, SEQ ID NO: 49, SEQ ID NO: 51, SEQ ID NO: 53, or SEQ ID NO: 55 may comprise 75 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 45, SEQ ID NO: 47, SEQ ID NO: 49, SEQ ID NO: 51, SEQ ID NO: 53, or SEQ ID NO: 55 may comprise 90 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 45, SEQ ID NO: 47, SEQ ID NO: 49, SEQ ID NO: 51, SEQ ID NO: 53, or SEQ ID NO: 55 may comprise 120 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 45, SEQ ID NO: 47, SEQ ID NO: 49, SEQ ID NO: 51, SEQ ID NO: 53, or SEQ ID NO: 55 may comprise 150 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 45, SEQ ID NO: 47, SEQ ID NO: 49, SEQ ID NO: 51, SEQ ID NO: 53, or SEQ ID NO: 55 may comprise 180 or more nucleotides, including a sequence encoding an immunodominant epitope.In some embodiments, a fragment of SEQ ID NO: 45, SEQ ID NO: 47, SEQ ID NO: 49, SEQ ID NO: 51, SEQ ID NO: 53, or SEQ ID NO: 55 may comprise 210 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 45, SEQ ID NO: 47, SEQ ID NO: 49, SEQ ID NO: 51, SEQ ID NO: 53, or SEQ ID NO: 55 may comprise 240 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 45, SEQ ID NO: 47, SEQ ID NO: 49, SEQ ID NO: 51, SEQ ID NO: 53, or SEQ ID NO: 55 may comprise 270 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 45, SEQ ID NO: 47, SEQ ID NO: 49, SEQ ID NO: 51, SEQ ID NO: 53, or SEQ ID NO: 55 may comprise 300 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 45, SEQ ID NO: 47, SEQ ID NO: 49, SEQ ID NO: 51, SEQ ID NO: 53, or SEQ ID NO: 55 may comprise 360 ​​or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 45, SEQ ID NO: 47, SEQ ID NO: 49, SEQ ID NO: 51, SEQ ID NO: 53, or SEQ ID NO: 55 may comprise 420 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 45, SEQ ID NO: 47, SEQ ID NO: 49, SEQ ID NO: 51, SEQ ID NO: 53, or SEQ ID NO: 55 may comprise 480 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 45, SEQ ID NO: 47, SEQ ID NO: 49, SEQ ID NO: 51, SEQ ID NO: 53, or SEQ ID NO: 55 may comprise 540 or more nucleotides, including a sequence encoding an immunodominant epitope.In some embodiments, a fragment of SEQ ID NO: 45, SEQ ID NO: 47, SEQ ID NO: 49, SEQ ID NO: 51, SEQ ID NO: 53, or SEQ ID NO: 55 may comprise 600 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 45, SEQ ID NO: 47, SEQ ID NO: 49, SEQ ID NO: 51, SEQ ID NO: 53, or SEQ ID NO: 55 may comprise 300 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 45, SEQ ID NO: 47, SEQ ID NO: 49, SEQ ID NO: 51, SEQ ID NO: 53, or SEQ ID NO: 55 may comprise 660 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 45, SEQ ID NO: 47, SEQ ID NO: 49, SEQ ID NO: 51, SEQ ID NO: 53, or SEQ ID NO: 55 may comprise 720 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 45, SEQ ID NO: 47, SEQ ID NO: 49, SEQ ID NO: 51, SEQ ID NO: 53, or SEQ ID NO: 55 may comprise 780 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 45, SEQ ID NO: 47, SEQ ID NO: 49, SEQ ID NO: 51, SEQ ID NO: 53, or SEQ ID NO: 55 may comprise 840 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 45, SEQ ID NO: 47, SEQ ID NO: 49, SEQ ID NO: 51, SEQ ID NO: 53, or SEQ ID NO: 55 may comprise 900 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 45, SEQ ID NO: 47, SEQ ID NO: 49, SEQ ID NO: 51, SEQ ID NO: 53, or SEQ ID NO: 55 may comprise 960 or more nucleotides, including a sequence encoding an immunodominant epitope.In some embodiments, a fragment of SEQ ID NO: 45, SEQ ID NO: 47, SEQ ID NO: 49, SEQ ID NO: 51, SEQ ID NO: 53, or SEQ ID NO: 55 may comprise 1020 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 45, SEQ ID NO: 47, SEQ ID NO: 49, SEQ ID NO: 51, SEQ ID NO: 53, or SEQ ID NO: 55 may comprise 1080 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 45, SEQ ID NO: 47, SEQ ID NO: 49, SEQ ID NO: 51, SEQ ID NO: 53, or SEQ ID NO: 55 may comprise 1140 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 45, SEQ ID NO: 47, SEQ ID NO: 49, SEQ ID NO: 51, SEQ ID NO: 53, or SEQ ID NO: 55 may comprise 1200 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 45, SEQ ID NO: 47, SEQ ID NO: 49, SEQ ID NO: 51, SEQ ID NO: 53, or SEQ ID NO: 55 may comprise 1260 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 45, SEQ ID NO: 47, SEQ ID NO: 49, SEQ ID NO: 51, SEQ ID NO: 53, or SEQ ID NO: 55 may comprise 1320 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 45, SEQ ID NO: 47, SEQ ID NO: 49, SEQ ID NO: 51, SEQ ID NO: 53, or SEQ ID NO: 55 may comprise 1380 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 45, SEQ ID NO: 47, SEQ ID NO: 49, SEQ ID NO: 51, SEQ ID NO: 53, or SEQ ID NO: 55 may comprise 1440 or more nucleotides, including a sequence encoding an immunodominant epitope.In some embodiments, a fragment of SEQ ID NO: 45, SEQ ID NO: 47, SEQ ID NO: 49, SEQ ID NO: 51, SEQ ID NO: 53, or SEQ ID NO: 55 may comprise 1500 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 45, SEQ ID NO: 47, SEQ ID NO: 49, SEQ ID NO: 51, SEQ ID NO: 53, or SEQ ID NO: 55 may comprise 1560 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 45, SEQ ID NO: 47, SEQ ID NO: 49, SEQ ID NO: 51, SEQ ID NO: 53, or SEQ ID NO: 55 may comprise 1620 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 45, SEQ ID NO: 47, SEQ ID NO: 49, SEQ ID NO: 51, SEQ ID NO: 53, or SEQ ID NO: 55 may comprise 1680 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 45, SEQ ID NO: 47, SEQ ID NO: 49, SEQ ID NO: 51, SEQ ID NO: 53, or SEQ ID NO: 55 may comprise 1740 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 45, SEQ ID NO: 47, SEQ ID NO: 49, SEQ ID NO: 51, SEQ ID NO: 53, or SEQ ID NO: 55 may comprise 1800 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 45, SEQ ID NO: 47, SEQ ID NO: 49, SEQ ID NO: 51, SEQ ID NO: 53, or SEQ ID NO: 55 may comprise 1860 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 45, SEQ ID NO: 47, SEQ ID NO: 49, SEQ ID NO: 51, SEQ ID NO: 53, or SEQ ID NO: 55 may comprise 1920 or more nucleotides, including a sequence encoding an immunodominant epitope.In some embodiments, a fragment of SEQ ID NO: 45, SEQ ID NO: 47, SEQ ID NO: 49, SEQ ID NO: 51, SEQ ID NO: 53, or SEQ ID NO: 55 may comprise 1980 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 45, SEQ ID NO: 47, SEQ ID NO: 49, SEQ ID NO: 51, SEQ ID NO: 53, or SEQ ID NO: 55 may comprise 2040 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 45, SEQ ID NO: 47, SEQ ID NO: 49, SEQ ID NO: 51, SEQ ID NO: 53, or SEQ ID NO: 55 may comprise 2100 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 45, SEQ ID NO: 47, SEQ ID NO: 49, SEQ ID NO: 51, SEQ ID NO: 53, or SEQ ID NO: 55 may comprise 2160 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 45, SEQ ID NO: 47, SEQ ID NO: 49, SEQ ID NO: 51, SEQ ID NO: 53, or SEQ ID NO: 55 may comprise 2220 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 45, SEQ ID NO: 47, SEQ ID NO: 49, SEQ ID NO: 51, SEQ ID NO: 53, or SEQ ID NO: 55 may comprise 2280 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 45, SEQ ID NO: 47, SEQ ID NO: 49, SEQ ID NO: 51, SEQ ID NO: 53, or SEQ ID NO: 55 may comprise 2340 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 45, SEQ ID NO: 47, SEQ ID NO: 49, SEQ ID NO: 51, SEQ ID NO: 53, or SEQ ID NO: 55 may comprise 2400 or more nucleotides, including a sequence encoding an immunodominant epitope.In some embodiments, a fragment of SEQ ID NO: 45, SEQ ID NO: 47, SEQ ID NO: 49, SEQ ID NO: 51, SEQ ID NO: 53, or SEQ ID NO: 55 may comprise 2460 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 45, SEQ ID NO: 47, SEQ ID NO: 49, SEQ ID NO: 51, SEQ ID NO: 53, or SEQ ID NO: 55 may comprise 2520 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 45, SEQ ID NO: 47, SEQ ID NO: 49, SEQ ID NO: 51, SEQ ID NO: 53, or SEQ ID NO: 55 may comprise 2580 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 45, SEQ ID NO: 47, SEQ ID NO: 49, SEQ ID NO: 51, SEQ ID NO: 53, or SEQ ID NO: 55 may comprise 2640 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 45, SEQ ID NO: 47, SEQ ID NO: 49, SEQ ID NO: 51, SEQ ID NO: 53, or SEQ ID NO: 55 may comprise 2700 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 45, SEQ ID NO: 47, SEQ ID NO: 49, SEQ ID NO: 51, SEQ ID NO: 53, or SEQ ID NO: 55 may comprise 2760 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 45, SEQ ID NO: 47, SEQ ID NO: 49, SEQ ID NO: 51, SEQ ID NO: 53, or SEQ ID NO: 55 may comprise 2820 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 45, SEQ ID NO: 47, SEQ ID NO: 49, SEQ ID NO: 51, SEQ ID NO: 53, or SEQ ID NO: 55 may comprise 2880 or more nucleotides, including a sequence encoding an immunodominant epitope.In some embodiments, a fragment of SEQ ID NO: 45, SEQ ID NO: 47, SEQ ID NO: 49, SEQ ID NO: 51, SEQ ID NO: 53, or SEQ ID NO: 55 may comprise 2940 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 45, SEQ ID NO: 47, SEQ ID NO: 49, SEQ ID NO: 51, SEQ ID NO: 53, or SEQ ID NO: 55 may comprise 3000 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 45, SEQ ID NO: 47, SEQ ID NO: 49, SEQ ID NO: 51, SEQ ID NO: 53, or SEQ ID NO: 55 may comprise 3060 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 45, SEQ ID NO: 47, SEQ ID NO: 49, SEQ ID NO: 51, SEQ ID NO: 53, or SEQ ID NO: 55 may comprise 3120 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 45, SEQ ID NO: 47, SEQ ID NO: 49, SEQ ID NO: 51, SEQ ID NO: 53, or SEQ ID NO: 55 may comprise 3180 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 45, SEQ ID NO: 47, SEQ ID NO: 49, SEQ ID NO: 51, SEQ ID NO: 53, or SEQ ID NO: 55 may comprise 3240 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 45, SEQ ID NO: 47, SEQ ID NO: 49, SEQ ID NO: 51, SEQ ID NO: 53, or SEQ ID NO: 55 may comprise 3300 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 45, SEQ ID NO: 47, SEQ ID NO: 49, SEQ ID NO: 51, SEQ ID NO: 53, or SEQ ID NO: 55 may comprise 3360 or more nucleotides, including a sequence encoding an immunodominant epitope.In some embodiments, a fragment of SEQ ID NO: 45, SEQ ID NO: 47, SEQ ID NO: 49, SEQ ID NO: 51, SEQ ID NO: 53, or SEQ ID NO: 55 may comprise 3420 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 45, SEQ ID NO: 47, SEQ ID NO: 49, SEQ ID NO: 51, SEQ ID NO: 53, or SEQ ID NO: 55 may comprise 3480 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, a fragment of SEQ ID NO: 45, SEQ ID NO: 47, SEQ ID NO: 49, SEQ ID NO: 51, SEQ ID NO: 53, or SEQ ID NO: 55 may comprise a sequence encoding an IgE leader sequence. In some embodiments, the fragment of SEQ ID NO:45, SEQ ID NO:47, SEQ ID NO:49, SEQ ID NO:51, SEQ ID NO:53, or SEQ ID NO:55 does not comprise the coding sequence for an IgE leader sequence.

[0150] A fragment may comprise less than 60 nucleotides, in some embodiments less than 75 nucleotides, in some embodiments less than 90 nucleotides, in some embodiments less than 120 nucleotides, in some embodiments less than 150 nucleotides, in some embodiments less than 180 nucleotides, in some embodiments less than 210 nucleotides, in some embodiments less than 240 nucleotides, in some embodiments less than 270 nucleotides, in some embodiments less than 300 nucleotides, in some embodiments less than 360 nucleotides, in some embodiments less than 420 nucleotides, in some embodiments less than 480 nucleotides, in some embodiments less than 540 nucleotides, in some embodiments less than 600 nucleotides. In some embodiments, less than 600 nucleotides, in some embodiments less than 660 nucleotides, in some embodiments less than 720 nucleotides, in some embodiments less than 780 nucleotides, in some embodiments less than 840 nucleotides, in some embodiments less than 900 nucleotides, in some embodiments less than 960 nucleotides, in some embodiments less than 1020 nucleotides, in some embodiments less than 1080 nucleotides, in some embodiments less than 1140 nucleotides, in some embodiments less than 1200 nucleotides, in some embodiments less than 1260 nucleotides, in some embodiments less than 1320 nucleotides, in some embodiments less than 1380 nucleotides. 0 nucleotides, in some embodiments less than 1440 nucleotides, in some embodiments less than 1500 nucleotides, in some embodiments less than 1560 nucleotides, in some embodiments less than 1620 nucleotides, in some embodiments less than 1680 nucleotides, in some embodiments less than 1740 nucleotides, in some embodiments less than 1800 nucleotides, in some embodiments less than 1860 nucleotides, in some embodiments less than 1920 nucleotides, in some embodiments less than 1980 nucleotides, in some embodiments less than 2040 nucleotides, in some embodiments less than 2100 nucleotides, in some embodiments less than 2160 nucleotides nucleotides, in some embodiments less than 2220 nucleotides, in some embodiments less than 2280 nucleotides, in some embodiments less than 2340 nucleotides, in some embodiments less than 2400 nucleotides, in some embodiments less than 2460 nucleotides, in some embodiments less than 2520 nucleotides, in some embodiments less than 2580 nucleotides, in some embodiments less than 2640 nucleotides, in some embodiments less than 2700 nucleotides, in some embodiments less than 2760 nucleotides, in some embodiments less than 2820 nucleotides, in some embodiments less than 2860 nucleotides, in some embodiments less than 2940 nucleotides,In some embodiments, less than 3000 nucleotides, in some embodiments, less than 3060 nucleotides, in some embodiments, less than 3120 nucleotides, in some embodiments, less than 3180 nucleotides, in some embodiments, less than 3240 nucleotides, in some embodiments, less than 3300 nucleotides, in some embodiments, less than 3360 nucleotides, in some embodiments, less than 3420 nucleotides, in some embodiments, less than 3480 nucleotides, and in some embodiments, less than 3510 nucleotides.

[0151] Fragments of SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, SEQ ID NO: 54, or SEQ ID NO: 56 may comprise 15 or more amino acids, including sequences comprising immunodominant epitopes. In some embodiments, fragments of SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, SEQ ID NO: 54, or SEQ ID NO: 56 may comprise 18 or more amino acids, including sequences comprising immunodominant epitopes. In some embodiments, fragments of SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, SEQ ID NO: 54, or SEQ ID NO: 56 may comprise 21 or more amino acids, including sequences comprising immunodominant epitopes. In some embodiments, a fragment of SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, SEQ ID NO: 54, or SEQ ID NO: 56 may comprise 24 or more amino acids, including sequences comprising immunodominant epitopes. In some embodiments, a fragment of SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, SEQ ID NO: 54, or SEQ ID NO: 56 may comprise 30 or more amino acids, including sequences comprising immunodominant epitopes. In some embodiments, a fragment of SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, SEQ ID NO: 54, or SEQ ID NO: 56 may comprise 36 or more amino acids, including sequences comprising immunodominant epitopes. In some embodiments, a fragment of SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, SEQ ID NO: 54, or SEQ ID NO: 56 may comprise 42 or more amino acids, including sequences comprising immunodominant epitopes. In some embodiments, a fragment of SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, SEQ ID NO: 54, or SEQ ID NO: 56 may comprise 48 or more amino acids, including sequences comprising immunodominant epitopes.In some embodiments, a fragment of SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, SEQ ID NO: 54, or SEQ ID NO: 56 may comprise 54 or more amino acids, including sequences comprising immunodominant epitopes. In some embodiments, a fragment of SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, SEQ ID NO: 54, or SEQ ID NO: 56 may comprise 60 or more amino acids, including sequences comprising immunodominant epitopes. In some embodiments, a fragment of SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, SEQ ID NO: 54, or SEQ ID NO: 56 may comprise 66 or more amino acids, including sequences comprising immunodominant epitopes. In some embodiments, a fragment of SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, SEQ ID NO: 54, or SEQ ID NO: 56 may comprise 72 or more amino acids, including sequences comprising immunodominant epitopes. In some embodiments, a fragment of SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, SEQ ID NO: 54, or SEQ ID NO: 56 may comprise 90 or more amino acids, including sequences comprising immunodominant epitopes. In some embodiments, a fragment of SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, SEQ ID NO: 54, or SEQ ID NO: 56 may comprise 120 or more amino acids, including sequences comprising immunodominant epitopes. In some embodiments, a fragment of SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, SEQ ID NO: 54, or SEQ ID NO: 56 may comprise 150 or more amino acids, including sequences comprising immunodominant epitopes. In some embodiments, a fragment of SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, SEQ ID NO: 54, or SEQ ID NO: 56 may comprise 180 or more amino acids, including sequences comprising immunodominant epitopes.In some embodiments, a fragment of SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, SEQ ID NO: 54, or SEQ ID NO: 56 may comprise 210 or more amino acids, including sequences comprising immunodominant epitopes. In some embodiments, a fragment of SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, SEQ ID NO: 54, or SEQ ID NO: 56 may comprise 240 or more amino acids, including sequences comprising immunodominant epitopes. In some embodiments, a fragment of SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, SEQ ID NO: 54, or SEQ ID NO: 56 may comprise 270 or more amino acids, including sequences comprising immunodominant epitopes. In some embodiments, a fragment of SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, SEQ ID NO: 54, or SEQ ID NO: 56 may comprise 300 or more amino acids, including sequences comprising immunodominant epitopes. In some embodiments, a fragment of SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, SEQ ID NO: 54, or SEQ ID NO: 56 may comprise 330 or more amino acids, including sequences comprising immunodominant epitopes. In some embodiments, a fragment of SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, SEQ ID NO: 54, or SEQ ID NO: 56 may comprise 360 ​​or more amino acids, including sequences comprising immunodominant epitopes. In some embodiments, a fragment of SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, SEQ ID NO: 54, or SEQ ID NO: 56 may comprise 390 or more amino acids, including sequences comprising immunodominant epitopes. In some embodiments, a fragment of SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, SEQ ID NO: 54, or SEQ ID NO: 56 may comprise 420 or more amino acids, including sequences comprising immunodominant epitopes.In some embodiments, a fragment of SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, SEQ ID NO: 54, or SEQ ID NO: 56 may comprise 450 or more amino acids, including sequences comprising immunodominant epitopes. In some embodiments, a fragment of SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, SEQ ID NO: 54, or SEQ ID NO: 56 may comprise 480 or more amino acids, including sequences comprising immunodominant epitopes. In some embodiments, a fragment of SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, SEQ ID NO: 54, or SEQ ID NO: 56 may comprise 510 or more amino acids, including sequences comprising immunodominant epitopes. In some embodiments, a fragment of SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, SEQ ID NO: 54, or SEQ ID NO: 56 may comprise 540 or more amino acids, including sequences comprising immunodominant epitopes. In some embodiments, a fragment of SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, SEQ ID NO: 54, or SEQ ID NO: 56 may comprise 570 or more amino acids, including sequences comprising immunodominant epitopes. In some embodiments, a fragment of SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, SEQ ID NO: 54, or SEQ ID NO: 56 may comprise 600 or more amino acids, including sequences comprising immunodominant epitopes. In some embodiments, a fragment of SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, SEQ ID NO: 54, or SEQ ID NO: 56 may comprise 630 or more amino acids, including sequences comprising immunodominant epitopes. In some embodiments, a fragment of SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, SEQ ID NO: 54, or SEQ ID NO: 56 may comprise 660 or more amino acids, including sequences comprising immunodominant epitopes.In some embodiments, a fragment of SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, SEQ ID NO: 54, or SEQ ID NO: 56 may comprise 690 or more amino acids, including sequences comprising immunodominant epitopes. In some embodiments, a fragment of SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, SEQ ID NO: 54, or SEQ ID NO: 56 may comprise 720 or more amino acids, including sequences comprising immunodominant epitopes. In some embodiments, a fragment of SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, SEQ ID NO: 54, or SEQ ID NO: 56 may comprise 750 or more amino acids, including sequences comprising immunodominant epitopes. In some embodiments, a fragment of SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, SEQ ID NO: 54, or SEQ ID NO: 56 may comprise 780 or more amino acids, including sequences comprising immunodominant epitopes. In some embodiments, a fragment of SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, SEQ ID NO: 54, or SEQ ID NO: 56 may comprise 810 or more amino acids, including sequences comprising immunodominant epitopes. In some embodiments, a fragment of SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, SEQ ID NO: 54, or SEQ ID NO: 56 may comprise 840 or more amino acids, including sequences comprising immunodominant epitopes. In some embodiments, a fragment of SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, SEQ ID NO: 54, or SEQ ID NO: 56 may comprise 870 or more amino acids, including sequences comprising immunodominant epitopes. In some embodiments, a fragment of SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, SEQ ID NO: 54, or SEQ ID NO: 56 may comprise 900 or more amino acids, including sequences comprising immunodominant epitopes.In some embodiments, a fragment of SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, SEQ ID NO: 54, or SEQ ID NO: 56 may comprise 930 or more amino acids, including sequences comprising immunodominant epitopes. In some embodiments, a fragment of SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, SEQ ID NO: 54, or SEQ ID NO: 56 may comprise 960 or more amino acids, including sequences comprising immunodominant epitopes. In some embodiments, a fragment of SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, SEQ ID NO: 54, or SEQ ID NO: 56 may comprise 990 or more amino acids, including sequences comprising immunodominant epitopes. In some embodiments, a fragment of SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, SEQ ID NO: 54, or SEQ ID NO: 56 may comprise 1020 or more amino acids, including sequences comprising immunodominant epitopes. In some embodiments, a fragment of SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, SEQ ID NO: 54, or SEQ ID NO: 56 may comprise 1050 or more amino acids, including sequences comprising immunodominant epitopes. In some embodiments, a fragment of SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, SEQ ID NO: 54, or SEQ ID NO: 56 may comprise 1080 or more amino acids, including sequences comprising immunodominant epitopes. In some embodiments, a fragment of SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, SEQ ID NO: 54, or SEQ ID NO: 56 may comprise 1110 or more amino acids, including sequences comprising immunodominant epitopes. In some embodiments, a fragment of SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, SEQ ID NO: 54, or SEQ ID NO: 56 may comprise 1140 or more amino acids, including sequences comprising immunodominant epitopes.In some embodiments, a fragment of SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, SEQ ID NO: 54, or SEQ ID NO: 56 may comprise 1170 or more amino acids, including sequences comprising immunodominant epitopes. In some embodiments, a fragment of SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, SEQ ID NO: 54, or SEQ ID NO: 56 may comprise 1200 or more amino acids, including sequences comprising immunodominant epitopes. In some embodiments, a fragment of SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, SEQ ID NO: 54, or SEQ ID NO: 56 may comprise 1230 or more amino acids, including sequences comprising immunodominant epitopes. In some embodiments, a fragment of SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, SEQ ID NO: 54, or SEQ ID NO: 56 may comprise 1260 or more amino acids, including sequences comprising immunodominant epitopes. In some embodiments, a fragment of SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, SEQ ID NO: 54, or SEQ ID NO: 56 may comprise 1290 or more amino acids, including sequences comprising immunodominant epitopes. In some embodiments, a fragment of SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, SEQ ID NO: 54, or SEQ ID NO: 56 may comprise 1320 or more amino acids, including sequences comprising immunodominant epitopes. In some embodiments, a fragment of SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, SEQ ID NO: 54, or SEQ ID NO: 56 may comprise 1350 or more amino acids, including sequences comprising immunodominant epitopes. In some embodiments, a fragment of SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, SEQ ID NO: 54, or SEQ ID NO: 56 may comprise 1380 or more amino acids, including sequences comprising immunodominant epitopes.In some embodiments, a fragment of SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, SEQ ID NO: 54, or SEQ ID NO: 56 may comprise 1410 or more amino acids, including sequences comprising immunodominant epitopes. In some embodiments, a fragment of SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, SEQ ID NO: 54, or SEQ ID NO: 56 may comprise 1440 or more amino acids, including sequences comprising immunodominant epitopes. In some embodiments, a fragment of SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, SEQ ID NO: 54, or SEQ ID NO: 56 may comprise 1470 or more amino acids, including sequences comprising immunodominant epitopes. In some embodiments, fragments of SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, SEQ ID NO: 54, or SEQ ID NO: 56 may comprise 1500 or more amino acids, including sequences comprising immunodominant epitopes. In some embodiments, fragments of SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, SEQ ID NO: 54, or SEQ ID NO: 56 may comprise the coding sequence of an IgE leader sequence. In some embodiments, fragments of SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, SEQ ID NO: 54, or SEQ ID NO: 56 do not comprise the coding sequence of an IgE leader sequence.

[0152] Fragments may comprise less than 24 amino acids, in some embodiments less than 30 amino acids, in some embodiments less than 36 amino acids, in some embodiments less than 42 amino acids, in some embodiments less than 48 amino acids, in some embodiments less than 54 amino acids, in some embodiments less than 60 amino acids, in some embodiments less than 72 amino acids, in some embodiments less than 90 amino acids, in some embodiments less than 120 amino acids, in some embodiments less than 150 amino acids, in some embodiments less than 180 amino acids, in some embodiments less than 210 amino acids, in some embodiments less than 240 amino acids, in some embodiments less than 260 amino acids, in some embodiments less than 290 amino acids, in some embodiments less than 320 amino acids, in some embodiments less than 350 amino acids, in some embodiments less than 380 amino acids, in some embodiments less than 410 amino acids, in some embodiments less than 440 amino acids, in some embodiments less than 470 amino acids, in some embodiments less than 500 amino acids, in some embodiments less than 530 amino acids, in some embodiments less than 560 amino acids, in some embodiments less than 590 amino acids, in some embodiments less than 620 amino acids, in some embodiments less than 640 amino acids, in some embodiments less than 660 amino acids, in some embodiments less than 670 amino acids, in some embodiments less than 680 amino acids, in some embodiments less than 700 amino acids, in some embodiments less than 710 amino acids, in some embodiments less than 720 amino acids, in some embodiments less than 730 amino acids, in some embodiments less than 740 amino acids, in some embodiments less than 750 amino acids, in some embodiments less than 760 amino acids, in some embodiments less than 770 amino acids, in some embodiments less than 780 amino acids, in some embodiments less than 790 amino acids, in some embodiments less than 800 amino acids, in some embodiments less than 810 amino acids, in some In some embodiments, the protein comprises less than 650 amino acids, in some embodiments less than 680 amino acids, in some embodiments less than 710 amino acids, in some embodiments less than 740 amino acids, in some embodiments less than 770 amino acids, in some embodiments less than 800 amino acids, in some embodiments less than 830 amino acids, in some embodiments less than 860 amino acids, in some embodiments less than 890 amino acids, in some embodiments less than 920 amino acids, in some embodiments less than 950 amino acids, in some embodiments less than 980 amino acids, in some embodiments less than 1010 amino acids, in some embodiments Less than 1040 amino acids, in some embodiments less than 1070 amino acids, in some embodiments less than 1200 amino acids, in some embodiments less than 1230 amino acids, in some embodiments less than 1260 amino acids, in some embodiments less than 1290 amino acids, in some embodiments less than 1320 amino acids, in some embodiments less than 1350 amino acids, in some embodiments less than 1380 amino acids, in some embodiments less than 1410 amino acids, in some embodiments less than 1440 amino acids, in some embodiments less than 1470 amino acids, and in some embodiments less than 1500 amino acids.

[0153] The present invention relates to anti-cancer immunogenic compositions. The immunogenic compositions may comprise one or more cancer antigens. The immunogenic compositions may prevent tumor growth. The immunogenic compositions may reduce tumor growth. The immunogenic compositions may prevent the metastasis of tumor cells. Depending on the cancer antigen, the immunogenic compositions may be targeted to treat cancers including, but not limited to, liver cancer, prostate cancer, melanoma, blood cancer, head and neck cancer, glioblastoma, recurrent respiratory papillomatosis, anal cancer, cervical cancer, and brain cancer.

[0154] The first step in developing immunogenic compositions is to identify cancer antigens that are not recognized by the immune system and are self-antigens. The identified cancer antigens are then converted from self-antigens to foreign antigens to be recognized by the immune system. Redesigning the nucleic acid and amino acid sequences of the recombinant cancer antigens from self-antigens to foreign antigens breaks the immune system's tolerance to the antigen. To break tolerance, several redesign strategies can be applied to cancer antigens, as described below.

[0155] The recombinant cancer antigens of the immunogenic composition are not recognized as self, thereby breaking tolerance. The breaking of tolerance can induce antigen-specific T cells and / or high-titer antibody responses, thereby inducing or triggering an immune response against a cancer or tumor expressing the antigen or reacting with the cancer or tumor. In some embodiments, the induced or triggered immune response can be a cellular immune response, a humoral immune response, or both a cellular and humoral immune response. In some embodiments, the induced or triggered cellular immune response can include the induction or secretion of interferon-γ (IFN-γ) and / or tumor necrosis factor α (TNF-α). In other embodiments, the induced or elicited immune response can reduce or inhibit one or more immunosuppressive factors that promote the growth of tumors or cancers expressing the antigen, such as, but not limited to, factors that downregulate MHC presentation, factors that upregulate antigen-specific regulatory T cells (Tregs), PD-L1, FasL, cytokines such as IL-10 and TFG-β, tumor-associated macrophages, tumor-associated fibroblasts, soluble factors produced by immunosuppressive cells, CTLA-4, PD-1, MDSC, MCP-1, and immune checkpoint molecules.

[0156] In certain embodiments, the immunogenic compositions can mediate the clearance of tumor cells or prevent their growth by inducing the following responses: (1) humoral immunity via B cell responses (to produce antibodies that block the production of monocyte chemoattractant protein-1 (MCP-1), thereby delaying the production of myeloid-derived suppressor cells (MDSC) and inhibiting tumor growth); (2) increasing the production of cytotoxic T lymphocytes such as CD8 +(CTL) to attack and kill tumor cells; (3) increase T helper cell responses; and (4) increase inflammatory responses via IFN-γ and TFN-α, or preferably a combination of the above responses. The immunogenic composition can increase tumor-free survival by 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, and 45%. The immunogenic compositions can reduce tumor mass by 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59% and 60% after immunization. The immunogenic compositions can prevent and block the increase of monocyte chemoattractant protein 1 (MCP-1), a cytokine secreted by myeloid-derived suppressor cells. The immunogenic compositions can increase tumor survival by 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59% and 60%.

[0157] The immunogenic composition can enhance the cellular immune response of a subject administered the immunogenic composition by about 50-fold to about 6000-fold, about 50-fold to about 5500-fold, about 50-fold to about 5000-fold, about 50-fold to about 4500-fold, about 100-fold to about 6000-fold, about 150-fold to about 6000-fold, about 200-fold to about 6000-fold, about 250-fold to about 6000-fold, or about 300-fold to about 6000-fold, compared to the cellular immune response of a subject not administered the immunogenic composition. In some embodiments, the immunogenic composition can enhance the cellular immune response of a subject to which the immunogenic composition is administered by about 50-fold, 100-fold, 150-fold, 200-fold, 250-fold, 300-fold, 350-fold, 400-fold, 450-fold, 500-fold, 550-fold, 600-fold, 650-fold, 700-fold, 750-fold, 800-fold, 850-fold, 900-fold, 950-fold, 1000-fold, 1100-fold, 1200-fold, 1300-fold, 1400-fold, 1500-fold, 1600-fold, 1700-fold, 1800-fold, 1900-fold, 2000-fold, 2100-fold, or more. times, 2200 times, 2300 times, 2400 times, 2500 times, 2600 times, 2700 times, 2800 times, 2900 times, 3000 times, 3100 times, 3200 times, 3300 times, 3400 times, 3500 times, 3600 times, 3700 times, 3800 times, 3900 times, 4000 times, 4100 times, 4200 times, 4300 times, 4400 times, 4500 times, 4600 times, 4700 times, 4800 times, 4900 times, 5000 times, 5100 times, 5200 times, 5300 times, 5400 times, 5500 times, 5600 times, 5700 times, 5800 times, 5900 times or 6000 times.

[0158] The immunogenic composition can increase interferon gamma (IFN-γ) levels in a subject administered the immunogenic composition by about 50-fold to about 6000-fold, about 50-fold to about 5500-fold, about 50-fold to about 5000-fold, about 50-fold to about 4500-fold, about 100-fold to about 6000-fold, about 150-fold to about 6000-fold, about 200-fold to about 6000-fold, about 250-fold to about 6000-fold, or about 300-fold to about 6000-fold, compared to IFN-γ levels in a subject not administered the immunogenic composition. In some embodiments, the immunogenic composition can increase the level of IFN-γ in a subject to which the immunogenic composition is administered by about 50-fold, 100-fold, 150-fold, 200-fold, 250-fold, 300-fold, 350-fold, 400-fold, 450-fold, 500-fold, 550-fold, 600-fold, 650-fold, 700-fold, 750-fold, 800-fold, 850-fold, 900-fold, 950-fold, 1000-fold, 1100-fold, 1200-fold, 1300-fold, 1400-fold, 1500-fold, 1600-fold, 1700-fold, 1800-fold, 1900-fold, 2000-fold, 2100-fold, or 3200-fold, compared to the level of IFN-γ in a subject not administered the immunogenic composition. 0 times, 2200 times, 2300 times, 2400 times, 2500 times, 2600 times, 2700 times, 2800 times, 2900 times, 3000 times, 3100 times, 3200 times, 3300 times, 3400 times, 3500 times, 3600 times, 3700 times, 3800 times, 3900 times, 4000 times, 4100 times, 4200 times, 4300 times, 4400 times, 4500 times, 4600 times, 4700 times, 4800 times, 4900 times, 5000 times, 5100 times, 5200 times, 5300 times, 5400 times, 5500 times, 5600 times, 5700 times, 5800 times, 5900 times or 6000 times.

[0159] The immunogenic composition can be a nucleic acid vaccine. In one embodiment, the nucleic acid vaccine is a DNA vaccine. DNA vaccines are disclosed in U.S. Patent Nos. 5,593,972, 5,739,118, 5,817,637, 5,830,876, 5,962,428, 5,981,505, 5,580,859, 5,703,055, and 5,676,594, which are incorporated herein by reference in their entirety. DNA vaccines may also contain elements or agents that inhibit their integration into chromosomes.

[0160] The immunogenic composition may be an RNA vaccine. The RNA vaccine may be introduced into cells. The RNA vaccine may also contain elements or agents that inhibit its integration into chromosomes.

[0161] The immunogenic composition can be a live attenuated vaccine, a vaccine using a recombinant vector to deliver the antigen, a subunit vaccine, and a glycoprotein vaccine, such as, but not limited to, U.S. Patent Nos. 4,510,245, 4,797,368, 4,722,848, 4,790,987, 4,920,209, 5,017,487, 5,077,044, 5,110,587, 5,112,749, 5,174,993, 5,223,424, 5,225,336, 5,240,703, 5,242,848, 5,294,441, 5,294,5 and 6,589,529 (each of which is incorporated herein by reference).

[0162] The immunogenic compositions of the present invention may have the properties required of an effective vaccine, such as safety, so that the vaccine itself does not cause disease or death; have a protective effect against disease; induce neutralizing antibodies; induce protective T cell responses; and provide ease of administration, few side effects, biostability, and low cost per dose. The immunogenic compositions may achieve some or all of these properties by containing cancer antigens as discussed below.

[0163] As described in more detail below, immunogenic compositions may also include one or more inhibitors (i.e., immune checkpoint inhibitors) of one or more immune checkpoint molecules. Immune checkpoint molecules are described in more detail below. Immune checkpoint inhibitors are any nucleic acid or protein that prevents any component in the immune system from presenting, presenting and / or differentiation in MHC class, T cells, presenting and / or differentiation, any cytokine, chemokine or signal transduction for immune cell proliferation and / or differentiation. As also described in more detail below, immunogenic compositions can be further combined with antibodies for checkpoint inhibitors such as PD-1, PDL-1, CTLA4, TIM3 and LAG3 to enhance the stimulation of cell and humoral immune response. Antibodies for immune checkpoint proteins are used to prevent immune checkpoint proteins from suppressing T cells and / or B cells from responding.

[0164] The immunogenic composition may also comprise an antigen or a fragment or variant thereof. The antigen may be any substance that induces an immune response in a subject. The antigen may be a nucleic acid sequence, an amino acid sequence, or a combination thereof. The nucleic acid sequence may be DNA, RNA, cDNA, variants thereof, fragments thereof, or combinations thereof. The nucleic acid sequence may also include additional sequences encoding linkers or tag sequences that are linked to the antigen via a peptide bond. The amino acid sequence may be a protein, a peptide, variants thereof, fragments thereof, or combinations thereof.

[0165] Antigens can be contained in proteins, nucleic acids or fragments thereof, or variants thereof, or combinations thereof, from a variety of organisms (e.g., viruses, parasites, bacteria, fungi or mammals). Antigens can be associated with autoimmune diseases, allergies or asthma. In other embodiments, antigens can be associated with cancer, herpes virus, influenza virus, hepatitis B virus, hepatitis C virus, human papillomavirus (HPV) or human immunodeficiency virus (HIV).

[0166] Some antigens can induce a strong immune response. Other antigens can induce a weak immune response. When combined with the TERT antigen, the antigen can trigger an even greater immune response.

[0167] a. Cancer antigens

[0168] The immunogenic composition may comprise one or more cancer antigens. The cancer antigen may be a nucleic acid sequence, an amino acid sequence, or a combination thereof. The nucleic acid sequence may be DNA, RNA, cDNA, variants thereof, fragments thereof, or combinations thereof. The nucleic acid sequence may also include additional sequences encoding linker or tag sequences linked to the cancer antigen via a peptide bond. The amino acid sequence may be a protein, a peptide, variants thereof, fragments thereof, or combinations thereof. The cancer antigen may be a recombinant cancer antigen.

[0169] In the present description, "tumor antigen" or "hyperproliferative disorder antigen" or "antigen associated with a hyperproliferative disorder" refers to an antigen that is common to a specific hyperproliferative disorder, such as cancer. The antigens discussed herein are included only as examples. This list is not intended to be exclusive, and additional examples will be apparent to those skilled in the art.

[0170] Cancer antigens or tumor antigens are proteins produced by tumor cells that elicit an immune response, particularly a T cell mediated immune response.The choice of antigen binding portion of the invention will depend on the particular type of cancer to be treated. Tumor antigens are well known in the art and include, for example, glioma-associated antigen, carcinoembryonic antigen (CEA), β-human chorionic gonadotropin, alpha-fetoprotein (AFP), lectin-reactive AFP, thyroglobulin, RAGE-1, MN-CAIX, human telomerase reverse transcriptase, RU1, RU2 (AS), intestinal carboxylesterase, mut hsp70-2, M-CSF, prostaglandinase, prostate-specific antigen (PSA), PAP, NY-ESO-1, LAGE-1a, p53, prostein, PSMA, Her2 / neu, survivin and telomerase, prostate cancer tumor antigen-1 (PCTA-1), MAGE, ELF2M, neutrophil elastase, ephrinB2, CD22, insulin growth factor (IGF)-I, IGF-II, IGF-I receptor, and mesothelin.

[0171] In one embodiment, tumor antigen comprises one or more antigenic cancer epitopes associated with malignant tumor. Malignant tumor expresses many kinds of proteins, and described protein can be used as target antigen of immune attack. These molecules include but are not limited to tissue-specific antigens such as MART-1, tyrosinase and GP100 in melanoma and prostate acid phosphatase (PAP) and prostate specific antigen (PSA) in prostate cancer. Other target molecules belong to the group of transformation-related molecules, such as oncogene HER-2 / Neu / ErbB-2. Another group of target antigens is carcinoembryonic antigens such as carcinoembryonic antigen (CEA). In B cell lymphoma, tumor-specific idiotype immunoglobulin constitutes true tumor-specific immunoglobulin antigen, which is unique for individual tumor. B cell differentiation antigens such as CD19, CD20 and CD37 are other candidates of target antigen in B cell lymphoma. Some of these antigens (CEA, HER-2, CD19, CD20, idiotype) have been used as targets of passive immunotherapy utilizing monoclonal antibodies, but with limited success.

[0172] The types of tumor antigens mentioned in the present invention may also be tumor-specific antigens (TSAs) or tumor-associated antigens (TAAs). TSAs are unique to tumor cells and are not present on other cells in the body. TAA-related antigens are not unique to tumor cells, but are on the contrary expressed on normal cells under conditions that cannot induce a state of immune tolerance to the antigen. The expression of antigens on tumors can occur under conditions that enable the immune system to respond to the antigen. TAAs may be antigens expressed on normal cells during fetal development when the immune system is immature and unable to respond, or they may be antigens that are usually present at very low levels in normal cells but expressed at much higher levels on tumor cells.

[0173] Non-limiting examples of TSA or TAA antigens include the following: differentiation antigens such as MART-1 / MelanA (MART-I), gp100 (Pmel 17), tyrosinase, TRP-1, TRP-2 and tumor-specific multi-lineage antigens such as: MAGE-1, MAGE-3, BAGE, GAGE-1, GAGE-2, p15; overexpressed embryonic antigens such as CEA; overexpressed oncogenes and mutated tumor suppressor genes such as p53, Ras, HER-2 / neu; unique tumor antigens caused by chromosomal translocations; such as BCR-ABL, E2A-PRL, H4-RET, IGH-IGK, MYL-RAR; and viral antigens such as Epstein-Barr virus antigen EBVA and human papillomavirus (HPV) antigens E6 and E7. Other large protein-based antigens include TSP-180, MAGE-4, MAGE-5, MAGE-6, RAGE, NY-ESO, p185erbB2, p180erbB-3, c-met, nm-23H1, PSA, TAG-72, CA 19-9, CA 72-4, CAM 17.1, NuMa, K-ras, β-catenin, CDK4, Mum-1, p 15, p 16, 43-9F, 5T4, 791Tgp72, alpha-fetoprotein, β-HCG, BCA225, BTAA, CA 125, CA 15-3\CA 27.29\BCAA, CA 195, CA 242, CA-50, CAM43, CD68\P1, CO-029, FGF-5, G250, Ga733\EpCAM, HTgp-175, M344, MA-50, MG7-Ag, MOV18, NB / 70K, NY-CO-1, RCAS1, SDCCAG16, TA-90\Mac-2 binding protein\cyclophilin C-related protein, TAAL6, TAG72, TLP and TPS.

[0174] The TERT antigen and optionally one or more antibodies targeting one or more immune checkpoint proteins can be combined or combined with a tumor antigen or a fragment or variant thereof. Cancer markers are known proteins that are present or upregulated relative to certain cancer cells. Cancer vaccines can be produced by methods that produce antigens representing such markers in a manner that breaks self-tolerance. Such cancer vaccines may include a TERT antigen and optionally one or more antibodies targeting one or more immune checkpoint proteins to enhance the immune response, and optionally one or more additional tumor antigens. The following are some exemplary tumor antigens:

[0175] (1) Tyrosinase (Tyr)

[0176] The immunogenic compositions of the present invention may comprise the cancer antigen tyrosinase (Tyr), a fragment thereof, or a variant thereof. Tyrosinase is a copper-containing enzyme with tyrosine hydroxylase and dopa oxidase catalytic activities that can be found in microorganisms as well as plant and animal tissues. Specifically, tyrosinase catalyzes the production of melanin and other pigments through the oxidation of phenols such as tyrosine. Mutations in the TYR gene lead to oculocutaneous albinism in mammals, and non-pathological polymorphisms of the TYR gene contribute to changes in skin pigmentation.

[0177] In addition, in cancer or tumor such as melanoma, tyrosinase can become imbalanced, thereby leading to increased melanin synthesis. Therefore, tyrosinase can be a cancer antigen associated with melanoma. In subjects suffering from melanoma, tyrosinase can be a target recognized by cytotoxic T cells. However, in some cases, the immune response for cancer or tumor (including melanoma) can be suppressed, thereby leading to supporting tumor formation and / or growth and thus the microenvironment of disease progression.

[0178] Immunosuppression can be promoted by myeloid-derived suppressor cells (MDSCs), which are a mixed population of immature macrophages, granulocytes, dendritic cells, and myeloid cells. The myeloid cells can be a heterogeneous population of bone marrow progenitor cells and immature myeloid cells (IMCs). Markers of MDSCs can include expression of Gr-1 and CD11b (i.e., Gr-1 + and CD11b + cell).

[0179] The circulation of MDSCs can be increased by chronic infection, and the expansion of MDSC populations can be associated with autoimmunity and inflammation. In particular, MDSC expansion (or presence in tumors or cancerous tissues) can promote tumor growth and evade immune detection and / or regulation. Therefore, MDSCs can affect the immune response to anti-cancer vaccines.

[0180] MDSCs can be regulated by regulators of G-protein signaling 2 (Rgs2), and Rgs2 can be highly expressed in tumor-derived MDSCs. Rgs2 can also be widely expressed in a variety of cells, such as myeloid cells. MDSCs derived from tumor-bearing mice can perform different functions than MDSCs derived from non-tumor-bearing mice. One such difference can be the upregulation of the production of the chemokine MCP-1 secreted by MDSCs. MCP-1 can promote cell migration through signal transduction via CCR2, a G protein-coupled receptor (GPCR) found on monocytes, endothelial cells, and T cells. Therefore, MCP-1 can cause the migration of endothelial cells, thereby promoting angiogenesis. Blocking MCP-1 via neutralizing antibodies can inhibit angiogenesis, thereby leading to reduced tumor metastasis and increased survival. Therefore, MCP-1 can be regarded as an angiogenic factor. In addition to secreting MCP-1, MDSCs can also secrete growth factors, thereby further promoting tumor growth.

[0181] The Tyr antigen can induce antigen-specific T cells and / or high-titer antibody responses, thereby inducing or triggering an immune response against a cancer or tumor expressing the antigen or reacting with the cancer or tumor. In some embodiments, the immune response induced or triggered can be a cellular, humoral, or cellular and humoral immune response. In some embodiments, the cellular immune response induced or triggered may include the induction or secretion of interferon-γ (IFN-γ) and / or tumor necrosis factor α (TNF-α). In other embodiments, the immune response induced or triggered can reduce or inhibit one or more immunosuppressive factors that promote the growth of tumors or cancers expressing the antigen, for example, but not limited to factors that downregulate MHC presentation, factors that upregulate antigen-specific regulatory T cells (Treg), PD-L1, FasL, cytokines such as IL-10 and TFG-β, tumor-associated macrophages, tumor-associated fibroblasts, soluble factors produced by immunosuppressive cells, CTLA-4, PD-1, MDSC, MCP-1, and immune checkpoint molecules, which are described in more detail below.

[0182] As shown herein, Tyr antigens induce antigen-specific T cells and high titer antibody responses against cancer cells or tumor cells (e.g., melanoma cells). In particular, Tyr antigens are important targets for immune-mediated clearance by inducing the following responses: (1) humoral immunity via B cell responses (to produce antibodies that block monocyte chemoattractant protein-1 (MCP-1) production, thereby delaying myeloid-derived suppressor cells (MDSC) and inhibiting tumor growth); (2) increasing the production of cytotoxic T lymphocytes such as CD8 +(CTL) to attack and kill tumor cells; (3) increase T helper cell response; and (4) increase inflammatory response via IFN-γ and TFN-α or a combination of the above responses. Thus, immunogenic compositions comprising Tyr antigens (e.g., shared Tyr antigens, which will be described in more detail below) provide a protective immune response against tumor formation and tumor growth because these immunogenic compositions prevent immunosuppression by reducing the population of MDSCs found in cancerous or tumor tissues and block the angiogenesis of cancerous or tumor tissues by reducing the production or secretion of MCP-1. Thus, any user can design the immunogenic compositions of the present invention comprising Tyr antigens to provide broad immunity against tumor formation, tumor metastasis, and tumor growth.

[0183] Tyr antigens may comprise protein epitopes that make them particularly effective as immunogens that can induce anti-Tyr immune responses against them. Tyr antigens may comprise full-length translation products, variants thereof, fragments thereof, or combinations thereof. Tyr antigens may comprise common proteins.

[0184] The nucleic acid sequence encoding the consensus Tyr antigen can be optimized for codon usage and corresponding RNA transcripts. The nucleic acid encoding the consensus Tyr antigen can be codons and RNA optimized for expression. In some embodiments, the nucleic acid sequence encoding the consensus Tyr antigen can include a Kozak sequence (e.g., GCC ACC) to enhance translation efficiency. The nucleic acid encoding the consensus Tyr antigen can include multiple stop codons (e.g., TGA TGA) to enhance translation efficiency.

[0185] The nucleic acid encoding the consensus Tyr antigen may also encode an immunoglobulin E (IgE) leader sequence. The nucleic acid encoding the consensus Tyr antigen may further encode an IgE leader sequence such that the amino acid sequence of the IgE leader sequence is linked to the amino acid sequence of the consensus Tyr antigen via a peptide bond. The nucleic acid encoding the consensus Tyr antigen may also include a nucleotide sequence encoding the IgE leader sequence. In some embodiments, the nucleic acid encoding the consensus Tyr antigen does not have or contains a nucleotide sequence encoding the IgE leader sequence.

[0186] The consensus Tyr antigen can be the nucleic acid sequence SEQ ID NO: 1, which encodes the amino acid sequence SEQ ID NO: 2. SEQ ID NO: 1 encodes the consensus Tyr protein linked to an IgE leader sequence. The consensus Tyr protein can be linked to an IgE leader sequence and an HA tag. In other embodiments, the consensus Tyr protein may not contain or be linked to an IgE leader sequence and / or an HA tag.

[0187] In some embodiments, the consensus Tyr antigen can be a nucleic acid sequence that is at least about 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the nucleic acid sequence shown in SEQ ID NO: 1. In other embodiments, the consensus Tyr antigen can be a nucleic acid sequence that encodes an amino acid sequence that is at least about 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence shown in SEQ ID NO: 2. The consensus Tyr antigen can be an amino acid sequence that is at least about 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical over the entire length of the amino acid sequence shown in SEQ ID NO:2.

[0188] Some embodiments relate to nucleic acid sequences encoding proteins homologous to the Tyr consensus protein, immunogenic fragments of the Tyr consensus protein, and immunogenic fragments of homologous proteins. Such nucleic acid molecules encoding immunogenic proteins having up to 95% homology to the consensus sequence, up to 96% homology to the consensus sequence, up to 97% homology to the consensus sequence, up to 98% homology to the consensus sequence, and up to 99% homology to the consensus sequence can be provided. Similarly, nucleic acid sequences encoding immunogenic fragments as described herein and immunogenic fragments of proteins homologous to the proteins described herein are also provided.

[0189] Some embodiments relate to nucleic acid molecules encoding immunogenic proteins that have 95% homology to the nucleic acid coding sequences herein. Some embodiments relate to nucleic acid molecules encoding immunogenic proteins that have 96% homology to the nucleic acid coding sequences herein. Some embodiments relate to nucleic acid molecules encoding immunogenic proteins that have 97% homology to the nucleic acid coding sequences herein. Some embodiments relate to nucleic acid molecules encoding immunogenic proteins that have 98% homology to the nucleic acid coding sequences herein. Some embodiments relate to nucleic acid molecules encoding immunogenic proteins that have 99% homology to the nucleic acid coding sequences herein. In some embodiments, the nucleic acid molecules having coding sequences disclosed herein that are homologous to the coding sequences of the consensus proteins disclosed herein include a sequence encoding an IgE leader sequence linked to the 5' end of the coding sequence encoding the homologous protein sequence disclosed herein.

[0190] Some embodiments relate to nucleic acid sequences encoding proteins having a specific percentage identity to a full-length Tyr consensus protein, immunogenic fragments of a Tyr consensus protein, and immunogenic fragments of a protein having identity to a Tyr consensus protein. Such nucleic acid molecules can be provided that encode immunogenic proteins having up to 80% identity to the full-length Tyr consensus sequence, up to 85% identity to the full-length Tyr consensus sequence, up to 90% identity to the full-length Tyr consensus sequence, up to 91% identity to the full-length Tyr consensus sequence, up to 92% identity to the full-length Tyr consensus sequence, up to 93% identity to the full-length Tyr consensus sequence, up to 94% identity to the full-length Tyr consensus sequence, up to 95% identity to the full-length Tyr consensus sequence, up to 96% identity to the full-length Tyr consensus sequence, up to 97% identity to the full-length Tyr consensus sequence, up to 98% identity to the full-length Tyr consensus sequence, and up to 99% identity to the full-length Tyr consensus sequence. Likewise, nucleic acid sequences encoding immunogenic fragments set forth herein and immunogenic fragments of proteins having similar percent identities to the Tyr proteins set forth herein as specified above are also provided.

[0191] In some embodiments, the nucleic acid sequence does not contain a coding sequence that encodes a leader sequence. In some embodiments, the nucleic acid sequence does not contain a coding sequence that encodes an IgE leader sequence.

[0192] Some embodiments relate to fragments of SEQ ID NO: 1. The fragments can be at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% of SEQ ID NO: 1. The fragments can have at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% homology to the fragments of SEQ ID NO: 1. The fragments can have at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to the fragments of SEQ ID NO: 1. In some embodiments, the fragment includes a sequence encoding a leader sequence, such as, for example, an immunoglobulin leader sequence, such as an IgE leader sequence. In some embodiments, the fragment does not contain a coding sequence encoding a leader sequence. In some embodiments, the fragment does not contain a coding sequence encoding a leader sequence, such as, for example, an IgE leader sequence.

[0193] In addition, the amino acid sequence of the consensus Tyr protein is SEQ ID NO: 2. The amino acid sequence of the consensus Tyr protein linked to the IgE leader sequence is SEQ ID NO: 2. The amino acid sequence of the consensus Tyr protein linked to the IgE leader sequence can be linked to an HA tag.

[0194] Some embodiments relate to proteins that are homologous to SEQ ID NO: 2. Some embodiments relate to immunogenic proteins that are 95% homologous to the consensus protein sequence shown in SEQ ID NO: 2. Some embodiments relate to immunogenic proteins that are 96% homologous to the consensus protein sequence shown in SEQ ID NO: 2. Some embodiments relate to immunogenic proteins that are 97% homologous to the consensus protein sequence shown in SEQ ID NO: 2. Some embodiments relate to immunogenic proteins that are 98% homologous to the consensus protein sequence shown in SEQ ID NO: 2. Some embodiments relate to immunogenic proteins that are 99% homologous to the consensus protein sequence shown in SEQ ID NO: 2.

[0195] Some embodiments relate to proteins identical to SEQ ID NO: 2. Some embodiments relate to immunogenic proteins having an amino acid sequence that is 80% identical to the full-length consensus amino acid sequence as shown in SEQ ID NO: 2. Some embodiments relate to immunogenic proteins having an amino acid sequence that is 85% identical to the full-length consensus amino acid sequence as shown in SEQ ID NO: 2. Some embodiments relate to immunogenic proteins having an amino acid sequence that is 90% identical to the full-length consensus amino acid sequence as shown in SEQ ID NO: 2. Some embodiments relate to immunogenic proteins having an amino acid sequence that is 91% identical to the full-length consensus amino acid sequence as shown in SEQ ID NO: 2. Some embodiments relate to immunogenic proteins having an amino acid sequence that is 92% identical to the full-length consensus amino acid sequence as shown in SEQ ID NO: 2. Some embodiments relate to immunogenic proteins having an amino acid sequence that is 93% identical to the full-length consensus amino acid sequence as shown in SEQ ID NO: 2. Some embodiments relate to immunogenic proteins having an amino acid sequence that is 94% identical to the full-length consensus amino acid sequence as shown in SEQ ID NO: 2. Some embodiments relate to immunogenic proteins having an amino acid sequence that is 95% identical to the full-length consensus amino acid sequence as shown in SEQ ID NO: 2. Some embodiments relate to immunogenic proteins having an amino acid sequence that is 96% identical to the full-length consensus amino acid sequence as shown in SEQ ID NO: 2. Some embodiments relate to immunogenic proteins having an amino acid sequence that is 97% identical to the full-length consensus amino acid sequence as shown in SEQ ID NO: 2. Some embodiments relate to immunogenic proteins having an amino acid sequence that is 98% identical to the full-length consensus amino acid sequence as shown in SEQ ID NO: 2. Some embodiments relate to immunogenic proteins having an amino acid sequence that is 99% identical to the full-length consensus amino acid sequence as shown in SEQ ID NO: 2.

[0196] In some embodiments, the protein does not contain a leader sequence. In some embodiments, the protein does not contain an IgE leader sequence. A fragment of the consensus protein may comprise at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% of the consensus protein. An immunogenic fragment of SEQ ID NO: 2 may be provided. The immunogenic fragment may comprise at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% of SEQ ID NO: 2. In some embodiments, the fragment includes a leader sequence, such as, for example, an immunoglobulin leader sequence, such as an IgE leader sequence. In some embodiments, the fragment does not contain a leader sequence. In some embodiments, the fragment does not contain a leader sequence, such as, for example, an IgE leader sequence.

[0197] Immunogenic fragments of proteins having amino acid sequences homologous to immunogenic fragments of SEQ ID NO: 2 can be provided. Such immunogenic fragments can comprise at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% of a protein having 95% or greater homology to SEQ ID NO: 2. Some embodiments relate to immunogenic fragments having 96% homology to immunogenic fragments of the consensus protein sequences herein. Some embodiments relate to immunogenic fragments having 97% homology to immunogenic fragments of the consensus protein sequences herein. Some embodiments relate to immunogenic fragments having 98% homology to immunogenic fragments of the consensus protein sequences herein. Some embodiments relate to immunogenic fragments having 99% homology to immunogenic fragments of the consensus protein sequences herein. In some embodiments, the fragments include a leader sequence, such as, for example, an immunoglobulin leader sequence, such as an IgE leader sequence. In some embodiments, the fragments do not contain a leader sequence. In some embodiments, the fragments do not contain a leader sequence, such as, for example, an IgE leader sequence.

[0198] Immunogenic fragments of proteins having an amino acid sequence identical to an immunogenic fragment of SEQ ID NO: 2 can be provided. Such immunogenic fragments can comprise at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% of a protein having 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to the amino acid sequence shown in SEQ ID NO: 2. In some embodiments, the fragments include a leader sequence, such as, for example, an immunoglobulin leader sequence, such as an IgE leader sequence. In some embodiments, the fragments do not contain a leader sequence. In some embodiments, the fragment does not contain a leader sequence, such as, for example, an IgE leader sequence.

[0199] As indicated herein with respect to linking a signal peptide or leader sequence to the N-terminus of a protein, the signal peptide / leader sequence replaces the protein's N-terminal methionine that is encoded by the start codon of the nucleic acid sequence encoding the protein without the signal peptide coding sequence.

[0200] (2) Tyrosinase-related protein 1 (TYRP1)

[0201] The immunogenic composition of the present invention may comprise the cancer antigen tyrosinase-related protein 1 (TYRP1), a fragment thereof or a variant thereof. TYRP1, encoded by the TYRP1 gene, is a 75 kDa transmembrane glycoprotein and is expressed in normal and malignant melanocytes and melanoma cells. Like tyrosinase, TYRP1 contains a modified (referred to as) M-box that can bind to microphthalmia transcription factor (MITF), which plays a central role in activating pigmentation, cell proliferation and differentiation within melanocytes. TYRP1 can help stabilize tyrosinase and can form heterodimers, which can prevent the premature death of melanocytes by attenuating tyrosinase-mediated cytotoxicity.

[0202] As described above for tyrosinase, tyrosinase-related protein 1 (TYRP-1) is also involved in the synthesis of melanin and pigment structures in melanocytes and can be recognized by the immune system of subjects with melanoma. Therefore, TYRP-1 can be an antigen associated with melanoma.

[0203] The TRYP-1 antigen can induce antigen-specific T cells and / or high-titer antibody responses, thereby inducing or triggering an immune response to a cancer or tumor expressing the antigen or reacting with the cancer or tumor. In some embodiments, the induced or triggered immune response can be a cellular immune response, a humoral immune response, or both a cellular and humoral immune response. In some embodiments, the induced or triggered cellular immune response can include the induction or secretion of interferon-γ (IFN-γ) and / or tumor necrosis factor α (TNF-α). In other embodiments, the induced or elicited immune response can reduce or inhibit one or more immunosuppressive factors that promote the growth of tumors or cancers expressing the antigen, such as, but not limited to, factors that downregulate MHC presentation, factors that upregulate antigen-specific regulatory T cells (Tregs), PD-L1, FasL, cytokines such as IL-10 and TFG-β, tumor-associated macrophages, tumor-associated fibroblasts, soluble factors produced by immunosuppressive cells, CTLA-4, PD-1, MDSC, MCP-1, and immune checkpoint molecules, which are described in more detail below.

[0204] TYRP-1 antigens may comprise protein epitopes that make them particularly effective as immunogens against which an anti-TYRP-1 immune response can be induced. TYRP-1 antigens may comprise full-length translation products, variants thereof, fragments thereof, or combinations thereof. TYRP-1 antigens may comprise consensus proteins.

[0205] The nucleic acid sequence encoding the consensus TYRP-1 antigen can be optimized for codon usage and corresponding RNA transcripts. The nucleic acid encoding the consensus TYRP-1 antigen can be codon and RNA optimized for expression. In some embodiments, the nucleic acid sequence encoding the consensus TYRP-1 antigen may include a Kozak sequence (e.g., GCC ACC) to enhance the efficiency of translation. The nucleic acid encoding the consensus TYRP-1 antigen may include multiple stop codons (e.g., TGA TGA) to enhance the efficiency of translation termination.

[0206] The nucleic acid encoding the consensus TYRP-1 antigen may also encode an immunoglobulin E (IgE) leader sequence. The nucleic acid encoding the consensus TYRP-1 antigen may further encode an IgE leader sequence such that the amino acid sequence of the IgE leader sequence is linked to the amino acid sequence of the consensus TYRP-1 antigen by a peptide bond. The nucleic acid encoding the consensus TYRP-1 antigen may also include a nucleotide sequence encoding the IgE leader sequence. In some embodiments, the nucleic acid encoding the consensus TYRP-1 antigen does not have or contains a nucleotide sequence encoding the IgE leader sequence.

[0207] The consensus TYRP-1 antigen can be the nucleic acid sequence of SEQ ID NO:3, which encodes the amino acid sequence of SEQ ID NO:4. SEQ ID NO:3 encodes the consensus TYRP-1 protein linked to an IgE leader sequence. The consensus TYRP-1 protein can be linked to an IgE leader sequence and an HA tag. In other embodiments, the consensus TYRP-1 protein may not contain or be linked to an IgE leader sequence and / or an HA tag.

[0208] In some embodiments, the consensus TYRP-1 antigen can be a nucleic acid sequence that is at least about 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical over the entire length of the nucleic acid sequence shown in SEQ ID NO: 3. In other embodiments, the consensus TYRP-1 antigen can be a nucleic acid sequence that encodes an amino acid sequence that is at least about 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical over the entire length of the amino acid sequence shown in SEQ ID NO: 4. The consensus TYRP-1 antigen can be an amino acid sequence that is at least about 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical over the entire length of the amino acid sequence shown in SEQ ID NO:4.

[0209] Some embodiments relate to nucleic acid sequences encoding proteins homologous to the TYRP-1 consensus protein, immunogenic fragments of the TYRP-1 consensus protein, and immunogenic fragments of homologous proteins. Such nucleic acid molecules encoding immunogenic proteins having up to 95% homology to the consensus sequence, up to 96% homology to the consensus sequence, up to 97% homology to the consensus sequence, up to 98% homology to the consensus sequence, and up to 99% homology to the consensus sequence can be provided. Similarly, nucleic acid sequences encoding immunogenic fragments as described herein and immunogenic fragments of proteins homologous to the proteins described herein are also provided.

[0210] Some embodiments relate to nucleic acid molecules encoding immunogenic proteins that have 95% homology to the nucleic acid coding sequences herein. Some embodiments relate to nucleic acid molecules encoding immunogenic proteins that have 96% homology to the nucleic acid coding sequences herein. Some embodiments relate to nucleic acid molecules encoding immunogenic proteins that have 97% homology to the nucleic acid coding sequences herein. Some embodiments relate to nucleic acid molecules encoding immunogenic proteins that have 98% homology to the nucleic acid coding sequences herein. Some embodiments relate to nucleic acid molecules encoding immunogenic proteins that have 99% homology to the nucleic acid coding sequences herein. In some embodiments, the nucleic acid molecules having coding sequences disclosed herein that are homologous to the coding sequences of the consensus proteins disclosed herein include a sequence encoding an IgE leader sequence linked to the 5' end of the coding sequence encoding the homologous protein sequence disclosed herein.

[0211] Some embodiments relate to nucleic acid sequences encoding proteins having a specific percent identity to a full-length TYRP-1 consensus protein, immunogenic fragments of a TYRP-1 consensus protein, and immunogenic fragments of a protein having identity to a TYRP-1 consensus protein. Such nucleic acid molecules can be provided that encode immunogenic proteins having up to 80% identity to the full-length TYRP-1 consensus sequence, up to 85% identity to the full-length TYRP-1 consensus sequence, up to 90% identity to the full-length TYRP-1 consensus sequence, up to 91% identity to the full-length TYRP-1 consensus sequence, up to 92% identity to the full-length TYRP-1 consensus sequence, up to 93% identity to the full-length TYRP-1 consensus sequence, up to 94% identity to the full-length TYRP-1 consensus sequence, up to 95% identity to the full-length TYRP-1 consensus sequence, up to 96% identity to the full-length TYRP-1 consensus sequence, up to 97% identity to the full-length TYRP-1 consensus sequence, up to 98% identity to the full-length TYRP-1 consensus sequence, and up to 99% identity to the full-length TYRP-1 consensus sequence. Likewise, nucleic acid sequences encoding immunogenic fragments set forth herein and immunogenic fragments of proteins having similar percent identities to the TYRP-1 proteins set forth herein as specified above are also provided.

[0212] In some embodiments, the nucleic acid sequence does not contain a coding sequence that encodes a leader sequence. In some embodiments, the nucleic acid sequence does not contain a coding sequence that encodes an IgE leader sequence.

[0213] Some embodiments relate to fragments of SEQ ID NO: 3. The fragments can be at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% of SEQ ID NO: 3. The fragments can have at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% homology to the fragments of SEQ ID NO: 3. The fragments can have at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to the fragments of SEQ ID NO: 3. In some embodiments, the fragment includes a sequence encoding a leader sequence, such as, for example, an immunoglobulin leader sequence, such as an IgE leader sequence. In some embodiments, the fragment does not contain a coding sequence encoding a leader sequence. In some embodiments, the fragment does not contain a coding sequence encoding a leader sequence, such as, for example, an IgE leader sequence.

[0214] In addition, the amino acid sequence of the consensus TYRP-1 protein is SEQ ID NO: 4. The amino acid sequence of the consensus TYRP-1 protein linked to the IgE leader sequence is SEQ ID NO: 4. The amino acid sequence of the consensus TYRP-1 protein linked to the IgE leader sequence can be linked to an HA tag.

[0215] Some embodiments relate to proteins homologous to SEQ ID NO: 4. Some embodiments relate to immunogenic proteins having 95% homology to the consensus protein sequence shown in SEQ ID NO: 4. Some embodiments relate to immunogenic proteins having 96% homology to the consensus protein sequence shown in SEQ ID NO: 4. Some embodiments relate to immunogenic proteins having 97% homology to the consensus protein sequence shown in SEQ ID NO: 4. Some embodiments relate to immunogenic proteins having 98% homology to the consensus protein sequence shown in SEQ ID NO: 4. Some embodiments relate to immunogenic proteins having 99% homology to the consensus protein sequence shown in SEQ ID NO: 4.

[0216] Some embodiments relate to a protein identical to SEQ ID NO: 4. Some embodiments relate to an immunogenic protein having an amino acid sequence that is 80% identical to the full-length consensus amino acid sequence as shown in SEQ ID NO: 4. Some embodiments relate to an immunogenic protein having an amino acid sequence that is 85% identical to the full-length consensus amino acid sequence as shown in SEQ ID NO: 4. Some embodiments relate to an immunogenic protein having an amino acid sequence that is 90% identical to the full-length consensus amino acid sequence as shown in SEQ ID NO: 4. Some embodiments relate to an immunogenic protein having an amino acid sequence that is 91% identical to the full-length consensus amino acid sequence as shown in SEQ ID NO: 4. Some embodiments relate to an immunogenic protein having an amino acid sequence that is 92% identical to the full-length consensus amino acid sequence as shown in SEQ ID NO: 4. Some embodiments relate to an immunogenic protein having an amino acid sequence that is 93% identical to the full-length consensus amino acid sequence as shown in SEQ ID NO: 4. Some embodiments relate to immunogenic proteins having an amino acid sequence that is 94% identical to the full-length consensus amino acid sequence as shown in SEQ ID NO: 4. Some embodiments relate to immunogenic proteins having an amino acid sequence that is 95% identical to the full-length consensus amino acid sequence as shown in SEQ ID NO: 4. Some embodiments relate to immunogenic proteins having an amino acid sequence that is 96% identical to the full-length consensus amino acid sequence as shown in SEQ ID NO: 4. Some embodiments relate to immunogenic proteins having an amino acid sequence that is 97% identical to the full-length consensus amino acid sequence as shown in SEQ ID NO: 4. Some embodiments relate to immunogenic proteins having an amino acid sequence that is 98% identical to the full-length consensus amino acid sequence as shown in SEQ ID NO: 4. Some embodiments relate to immunogenic proteins having an amino acid sequence that is 99% identical to the full-length consensus amino acid sequence as shown in SEQ ID NO: 4.

[0217] In some embodiments, the protein does not contain a leader sequence. In some embodiments, the protein does not contain an IgE leader sequence. A fragment of the consensus protein may comprise at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% of the consensus TYRP-1 protein. An immunogenic fragment of SEQ ID NO: 4 may be provided. The immunogenic fragment may comprise at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% of SEQ ID NO: 4. In some embodiments, the fragment includes a leader sequence, such as, for example, an immunoglobulin leader sequence, such as an IgE leader sequence. In some embodiments, the fragment does not contain a leader sequence. In some embodiments, the fragment does not contain a leader sequence, such as, for example, an IgE leader sequence.

[0218] Immunogenic fragments of proteins having amino acid sequences homologous to immunogenic fragments of SEQ ID NO: 4 can be provided. Such immunogenic fragments can comprise at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% of a protein having 95% or greater homology to SEQ ID NO: 4. Some embodiments relate to immunogenic fragments having 96% homology to immunogenic fragments of the consensus protein sequences herein. Some embodiments relate to immunogenic fragments having 97% homology to immunogenic fragments of the consensus protein sequences herein. Some embodiments relate to immunogenic fragments having 98% homology to immunogenic fragments of the consensus protein sequences herein. Some embodiments relate to immunogenic fragments having 99% homology to immunogenic fragments of the consensus protein sequences herein. In some embodiments, the fragments include a leader sequence, such as, for example, an immunoglobulin leader sequence, such as an IgE leader sequence. In some embodiments, the fragments do not contain a leader sequence. In some embodiments, the fragments do not contain a leader sequence, such as, for example, an IgE leader sequence.

[0219] Immunogenic fragments of proteins having an amino acid sequence identical to an immunogenic fragment of SEQ ID NO: 4 can be provided. Such immunogenic fragments can comprise at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% of a protein having 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to the amino acid sequence shown in SEQ ID NO: 4. In some embodiments, the fragments include a leader sequence, such as, for example, an immunoglobulin leader sequence, such as an IgE leader sequence. In some embodiments, the fragments do not contain a leader sequence. In some embodiments, the fragment does not contain a leader sequence, such as, for example, an IgE leader sequence.

[0220] As indicated herein with respect to linking a signal peptide or leader sequence to the N-terminus of a protein, the signal peptide / leader sequence replaces the protein's N-terminal methionine that is encoded by the start codon of the nucleic acid sequence encoding the protein without the signal peptide coding sequence.

[0221] (3) Tyrosinase-related protein 2 (TYRP2)

[0222] The immunogenic compositions of the present invention may comprise the cancer antigen tyrosinase-related protein 2 (TYRP2; also known as dopachrome tautomerase (DCT)), a fragment thereof, or a variant thereof. TYRP2 / DCT, encoded by the TYRP2 / DCT gene, is a 519-amino acid protein expressed in normal and malignant melanocytes and melanoma cells. TYRP2 / DCT is a well-characterized melanocyte-specific enzyme that binds to tyrosinase and TYRP1 in melanocytes and plays a role in the conversion of L-tyrosine to melanin. DCT specifically catalyzes the tautomerization of the melanin precursor L-dopachrome to 5,6-dihydroindole-2-carboxylic acid (DHICA), which is subsequently oxidized by TYRP1 (as discussed above) to form eumelanin. Studies have shown that TYRP2 / DCT may be a mediator of drug resistance in melanoma cells, with specificity for DNA-damaging agents. Since TYRP2 / DCT is frequently reported to be highly expressed in melanoma, this melanocyte-specific enzyme plays an important role in contributing to the intrinsic resistance phenotype of melanoma to various anticancer DNA-damaging drugs.

[0223] As described above for tyrosinase, tyrosinase-related protein 2 (TYRP-2) is also involved in the synthesis of melanin and can be recognized by the immune system of subjects with melanoma. In addition, TYRP-2 can mediate drug resistance in melanoma cells. Therefore, TYRP-2 can be an antigen associated with melanoma.

[0224] The TRYP-2 antigen can induce antigen-specific T cells and / or high-titer antibody responses, thereby inducing or triggering an immune response to a cancer or tumor expressing the antigen or reacting with the cancer or tumor. In some embodiments, the induced or triggered immune response can be a cellular immune response, a humoral immune response, or both a cellular and humoral immune response. In some embodiments, the induced or triggered cellular immune response can include the induction or secretion of interferon-γ (IFN-γ) and / or tumor necrosis factor α (TNF-α). In other embodiments, the induced or elicited immune response can reduce or inhibit one or more immunosuppressive factors that promote the growth of tumors or cancers expressing the antigen, such as, but not limited to, factors that downregulate MHC presentation, factors that upregulate antigen-specific regulatory T cells (Tregs), PD-L1, FasL, cytokines such as IL-10 and TFG-β, tumor-associated macrophages, tumor-associated fibroblasts, soluble factors produced by immunosuppressive cells, CTLA-4, PD-1, MDSC, MCP-1, and immune checkpoint molecules, which are described in more detail below.

[0225] TYRP2 antigens may comprise protein epitopes that make them particularly effective as immunogens against which an anti-TYRP2 immune response can be induced. TYRP2 antigens may comprise full-length translation products, variants thereof, fragments thereof, or combinations thereof. TYRP2 antigens may comprise a consensus protein.

[0226] The nucleic acid sequence encoding the consensus TYRP2 antigen can be optimized for codon usage and corresponding RNA transcripts. The nucleic acid encoding the consensus TYRP2 antigen can be codons and RNA optimized for expression. In some embodiments, the nucleic acid sequence encoding the consensus TYRP2 antigen may include a Kozak sequence (e.g., GCC ACC) to enhance the efficiency of translation. The nucleic acid encoding the consensus TYRP2 antigen may include multiple stop codons (e.g., TGA TGA) to enhance the efficiency of translation termination.

[0227] The nucleic acid encoding the consensus TYRP2 antigen may also encode an immunoglobulin E (IgE) leader sequence. The nucleic acid encoding the consensus TYRP2 antigen may further encode an IgE leader sequence such that the amino acid sequence of the IgE leader sequence is linked to the amino acid sequence of the consensus TYRP2 antigen by a peptide bond. The nucleic acid encoding the consensus TYRP2 antigen may also include a nucleotide sequence encoding the IgE leader sequence. In some embodiments, the nucleic acid encoding the consensus TYRP2 antigen does not have or contains a nucleotide sequence encoding the IgE leader sequence.

[0228] The consensus TYRP2 antigen can be the nucleic acid sequence SEQ ID NO: 5, which encodes the amino acid sequence SEQ ID NO: 6. SEQ ID NO: 5 encodes the consensus TYRP2 protein linked to an IgE leader sequence. The consensus TYRP2 protein can be linked to an IgE leader sequence and an HA tag. In other embodiments, the consensus TYRP2 protein may not contain or be linked to an IgE leader sequence and / or an HA tag.

[0229] In some embodiments, the consensus TYRP2 antigen can be a nucleic acid sequence that is at least about 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the nucleic acid sequence shown in SEQ ID NO: 5. In other embodiments, the consensus TYRP2 antigen can be a nucleic acid sequence that encodes an amino acid sequence that is at least about 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence shown in SEQ ID NO: 6. The consensus TYRP2 antigen can be an amino acid sequence that is at least about 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical over the entire length of the amino acid sequence shown in SEQ ID NO:6.

[0230] Some embodiments relate to nucleic acid sequences encoding proteins homologous to the TYRP2 consensus protein, immunogenic fragments of the TYRP2 consensus protein, and immunogenic fragments of homologous proteins. Such nucleic acid molecules encoding immunogenic proteins having up to 95% homology to the consensus sequence, up to 96% homology to the consensus sequence, up to 97% homology to the consensus sequence, up to 98% homology to the consensus sequence, and up to 99% homology to the consensus sequence can be provided. Similarly, nucleic acid sequences encoding immunogenic fragments as described herein and immunogenic fragments of proteins homologous to the proteins described herein are also provided.

[0231] Some embodiments relate to nucleic acid molecules encoding immunogenic proteins that have 95% homology to the nucleic acid coding sequences herein. Some embodiments relate to nucleic acid molecules encoding immunogenic proteins that have 96% homology to the nucleic acid coding sequences herein. Some embodiments relate to nucleic acid molecules encoding immunogenic proteins that have 97% homology to the nucleic acid coding sequences herein. Some embodiments relate to nucleic acid molecules encoding immunogenic proteins that have 98% homology to the nucleic acid coding sequences herein. Some embodiments relate to nucleic acid molecules encoding immunogenic proteins that have 99% homology to the nucleic acid coding sequences herein. In some embodiments, the nucleic acid molecules having coding sequences disclosed herein that are homologous to the coding sequences of the consensus proteins disclosed herein include a sequence encoding an IgE leader sequence linked to the 5' end of the coding sequence encoding the homologous protein sequence disclosed herein.

[0232] Some embodiments relate to nucleic acid sequences encoding proteins having a specific percent identity to a full-length TYRP2 consensus protein, immunogenic fragments of a TYRP2 consensus protein, and immunogenic fragments of a protein having identity to a TYRP2 consensus protein. Such nucleic acid molecules can be provided that encode immunogenic proteins having up to 80% identity to the full-length TYRP2 consensus sequence, up to 85% identity to the full-length consensus sequence, up to 90% identity to the full-length TYRP2 consensus sequence, up to 91% identity to the full-length TYRP2 consensus sequence, up to 92% identity to the full-length TYRP2 consensus sequence, up to 93% identity to the full-length TYRP2 consensus sequence, up to 94% identity to the full-length TYRP2 consensus sequence, up to 95% identity to the full-length TYRP2 consensus sequence, up to 96% identity to the full-length TYRP2 consensus sequence, up to 97% identity to the full-length TYRP2 consensus sequence, up to 98% identity to the full-length TYRP2 consensus sequence, and up to 99% identity to the full-length TYRP2 consensus sequence. Likewise, nucleic acid sequences encoding immunogenic fragments set forth herein and immunogenic fragments of proteins having similar percent identities to the TYRP2 proteins set forth herein as specified above are also provided.

[0233] In some embodiments, the nucleic acid sequence does not contain a coding sequence that encodes a leader sequence. In some embodiments, the nucleic acid sequence does not contain a coding sequence that encodes an IgE leader sequence.

[0234] Some embodiments relate to fragments of SEQ ID NO: 5. The fragments can be at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% of SEQ ID NO: 5. The fragments can have at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% homology to the fragments of SEQ ID NO: 5. The fragments can have at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to the fragments of SEQ ID NO: 5. In some embodiments, the fragment includes a sequence encoding a leader sequence, such as, for example, an immunoglobulin leader sequence, such as an IgE leader sequence. In some embodiments, the fragment does not contain a coding sequence encoding a leader sequence. In some embodiments, the fragment does not contain a coding sequence encoding a leader sequence, such as, for example, an IgE leader sequence.

[0235] In addition, the amino acid sequence of the consensus TYRP2 protein is SEQ ID NO: 6. The amino acid sequence of the consensus TYRP2 protein linked to the IgE leader sequence is SEQ ID NO: 6. The amino acid sequence of the consensus TYRP2 protein linked to the IgE leader sequence can be linked to an HA tag.

[0236] Some embodiments relate to proteins homologous to SEQ ID NO: 6. Some embodiments relate to immunogenic proteins having 95% homology to the consensus protein sequence shown in SEQ ID NO: 6. Some embodiments relate to immunogenic proteins having 96% homology to the consensus protein sequence shown in SEQ ID NO: 6. Some embodiments relate to immunogenic proteins having 97% homology to the consensus protein sequence shown in SEQ ID NO: 6. Some embodiments relate to immunogenic proteins having 98% homology to the consensus protein sequence shown in SEQ ID NO: 6. Some embodiments relate to immunogenic proteins having 99% homology to the consensus protein sequence shown in SEQ ID NO: 6.

[0237] Some embodiments relate to proteins identical to SEQ ID NO: 6. Some embodiments relate to immunogenic proteins having an amino acid sequence that is 80% identical to the full-length consensus amino acid sequence as shown in SEQ ID NO: 6. Some embodiments relate to immunogenic proteins having an amino acid sequence that is 85% identical to the full-length consensus amino acid sequence as shown in SEQ ID NO: 6. Some embodiments relate to immunogenic proteins having an amino acid sequence that is 90% identical to the full-length consensus amino acid sequence as shown in SEQ ID NO: 6. Some embodiments relate to immunogenic proteins having an amino acid sequence that is 91% identical to the full-length consensus amino acid sequence as shown in SEQ ID NO: 6. Some embodiments relate to immunogenic proteins having an amino acid sequence that is 92% identical to the full-length consensus amino acid sequence as shown in SEQ ID NO: 6. Some embodiments relate to immunogenic proteins having an amino acid sequence that is 93% identical to the full-length consensus amino acid sequence as shown in SEQ ID NO: 6. Some embodiments relate to immunogenic proteins having an amino acid sequence that is 94% identical to the full-length consensus amino acid sequence as shown in SEQ ID NO: 6. Some embodiments relate to immunogenic proteins having an amino acid sequence that is 95% identical to the full-length consensus amino acid sequence as shown in SEQ ID NO: 6. Some embodiments relate to immunogenic proteins having an amino acid sequence that is 96% identical to the full-length consensus amino acid sequence as shown in SEQ ID NO: 6. Some embodiments relate to immunogenic proteins having an amino acid sequence that is 97% identical to the full-length consensus amino acid sequence as shown in SEQ ID NO: 6. Some embodiments relate to immunogenic proteins having an amino acid sequence that is 98% identical to the full-length consensus amino acid sequence as shown in SEQ ID NO: 6. Some embodiments relate to immunogenic proteins having an amino acid sequence that is 99% identical to the full-length consensus amino acid sequence as shown in SEQ ID NO: 6.

[0238] In some embodiments, the protein does not contain a leader sequence. In some embodiments, the protein does not contain an IgE leader sequence. A fragment of the consensus protein can comprise at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% of the consensus protein. An immunogenic fragment of SEQ ID NO: 6 can be provided. The immunogenic fragment may comprise at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% of SEQ ID NO: 6. In some embodiments, the fragment includes a leader sequence, such as, for example, an immunoglobulin leader sequence, such as an IgE leader sequence. In some embodiments, the fragment does not contain a leader sequence. In some embodiments, the fragment does not contain a leader sequence, such as, for example, an IgE leader sequence.

[0239] Immunogenic fragments of proteins having amino acid sequences homologous to immunogenic fragments of SEQ ID NO: 6 can be provided. Such immunogenic fragments can comprise at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% of a protein having 95% or greater homology to SEQ ID NO: 6. Some embodiments relate to immunogenic fragments having 96% homology to immunogenic fragments of the consensus protein sequences herein. Some embodiments relate to immunogenic fragments having 97% homology to immunogenic fragments of the consensus protein sequences herein. Some embodiments relate to immunogenic fragments having 98% homology to immunogenic fragments of the consensus protein sequences herein. Some embodiments relate to immunogenic fragments having 99% homology to immunogenic fragments of the consensus protein sequences herein. In some embodiments, the fragments include a leader sequence, such as, for example, an immunoglobulin leader sequence, such as an IgE leader sequence. In some embodiments, the fragments do not contain a leader sequence. In some embodiments, the fragments do not contain a leader sequence, such as, for example, an IgE leader sequence.

[0240] Immunogenic fragments of proteins having an amino acid sequence identical to an immunogenic fragment of SEQ ID NO: 6 can be provided. Such immunogenic fragments can comprise at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% of a protein having 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to the amino acid sequence shown in SEQ ID NO: 6. In some embodiments, the fragments include a leader sequence, such as, for example, an immunoglobulin leader sequence, such as an IgE leader sequence. In some embodiments, the fragments do not contain a leader sequence. In some embodiments, the fragment does not contain a leader sequence, such as, for example, an IgE leader sequence.

[0241] As indicated herein with respect to linking a signal peptide or leader sequence to the N-terminus of a protein, the signal peptide / leader sequence replaces the protein's N-terminal methionine that is encoded by the start codon of the nucleic acid sequence encoding the protein without the signal peptide coding sequence.

[0242] (4) Melanoma-associated antigen 4 (MAGEA4)

[0243] The immunogenic compositions of the present invention may comprise the cancer antigen melanoma-associated antigen 4 (MAGEA4), a fragment thereof, or a variant thereof. MAGE-A4, encoded by the MAGE-A4 gene, is a 317-amino acid protein expressed in male germ cells and in tumor cells of various histological types, such as gastrointestinal, esophageal, and lung cancers. MAGE-A4 binds to the oncoprotein Gankyrin. This specific binding of MAGE-A4 is mediated by its C-terminus. Studies have shown that exogenous MAGE-A4 can partially inhibit the anchorage-independent growth of cells overexpressing Gankyrin in vitro and inhibit the formation of migratory tumors from these cells in nude mice. This inhibition is dependent on the binding between MAGE-A4 and Gankyrin, suggesting that the interaction between Gankyrin and MAGE-A4 inhibits Gankyrin-mediated carcinogenesis. MAGE expression in tumor tissue may not be a cause, but rather a consequence of tumorigenesis, and that MAGE genes participate in immune processes by targeting early-stage tumor cells for destruction.

[0244] Melanoma-associated antigen 4 (MAGEA4) protein may be involved in embryonic development as well as tumor transformation and / or progression. MAGEA4 is normally expressed in the testis and placenta. However, MAGEA4 can be expressed in many different types of tumors, such as melanoma, head and neck squamous cell carcinoma, lung cancer, and breast cancer. Therefore, MAGEA4 may be an antigen associated with a variety of tumors.

[0245] The MAGEA4 antigen can induce antigen-specific T cells and / or high-titer antibody responses, thereby inducing or triggering an immune response to a cancer or tumor expressing the antigen or to the cancer or tumor reaction. In some embodiments, the induced or triggered immune response can be a cellular immune response, a humoral immune response, or both a cellular and humoral immune response. In some embodiments, the induced or triggered cellular immune response can include the induction or secretion of interferon-γ (IFN-γ) and / or tumor necrosis factor α (TNF-α). In other embodiments, the induced or triggered immune response can reduce or inhibit one or more immunosuppressive factors that promote the growth of tumors or cancers expressing the antigen, for example, but not limited to factors that downregulate MHC presentation, factors that upregulate antigen-specific regulatory T cells (Treg), PD-L1, FasL, cytokines such as IL-10 and TFG-β, tumor-associated macrophages, tumor-associated fibroblasts, soluble factors produced by immunosuppressive cells, CTLA-4, PD-1, MDSC, MCP-1, and immune checkpoint molecules, which are described in more detail below.

[0246] MAGEA4 antigens may comprise protein epitopes that make them particularly effective as immunogens against which an anti-MAGEA4 immune response can be induced. MAGEA4 antigens may comprise full-length translation products, variants thereof, fragments thereof, or combinations thereof. MAGEA4 antigens may comprise a consensus protein.

[0247] The nucleic acid sequence encoding the consensus MAGEA4 antigen can be optimized for codon usage and corresponding RNA transcripts. The nucleic acid encoding the consensus MAGEA4 antigen can be codon and RNA optimized for expression. In some embodiments, the nucleic acid sequence encoding the consensus MAGEA4 antigen can include a Kozak sequence (e.g., GCC ACC) to enhance translation efficiency. The nucleic acid encoding the consensus MAGEA4 antigen can include multiple stop codons (e.g., TGA TGA) to enhance translation termination efficiency.

[0248] The nucleic acid encoding the consensus MAGEA4 antigen may also encode an immunoglobulin E (IgE) leader sequence. The nucleic acid encoding the consensus Tyr antigen may further encode an IgE leader sequence such that the amino acid sequence of the IgE leader sequence is linked to the amino acid sequence of the consensus MAGEA4 antigen via a peptide bond. The nucleic acid encoding the consensus MAGEA4 antigen may also include a nucleotide sequence encoding the IgE leader sequence. In some embodiments, the nucleic acid encoding the consensus MAGEA4 antigen does not have or contains a nucleotide sequence encoding the IgE leader sequence.

[0249] The consensus MAGEA4 antigen can be the nucleic acid sequence SEQ ID NO:7, which encodes the amino acid sequence SEQ ID NO:8. SEQ ID NO:7 encodes the consensus MAGEA4 protein linked to an IgE leader sequence. The consensus MAGEA4 protein can be linked to an IgE leader sequence and an HA tag. In other embodiments, the consensus MAGEA4 protein may not contain or be linked to an IgE leader sequence and / or an HA tag.

[0250] In some embodiments, the consensus MAGEA4 antigen can be a nucleic acid sequence that is at least about 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the nucleic acid sequence shown in SEQ ID NO: 7. In other embodiments, the consensus MAGEA4 antigen can be a nucleic acid sequence that encodes an amino acid sequence that is at least about 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence shown in SEQ ID NO: 8. The consensus MAGEA4 antigen can be an amino acid sequence that is at least about 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical over the entire length of the amino acid sequence shown in SEQ ID NO:8.

[0251] Some embodiments relate to nucleic acid sequences encoding proteins homologous to the MAGEA4 consensus protein, immunogenic fragments of the MAGEA4 consensus protein, and immunogenic fragments of homologous proteins. Such nucleic acid molecules can be provided that encode immunogenic proteins having up to 95% homology to the consensus sequence, up to 96% homology to the consensus sequence, up to 97% homology to the consensus sequence, up to 98% homology to the consensus sequence, and up to 99% homology to the consensus sequence. Similarly, nucleic acid sequences encoding immunogenic fragments as described herein and immunogenic fragments of proteins homologous to the proteins described herein are also provided.

[0252] Some embodiments relate to nucleic acid molecules encoding immunogenic proteins that have 95% homology to the nucleic acid coding sequences herein. Some embodiments relate to nucleic acid molecules encoding immunogenic proteins that have 96% homology to the nucleic acid coding sequences herein. Some embodiments relate to nucleic acid molecules encoding immunogenic proteins that have 97% homology to the nucleic acid coding sequences herein. Some embodiments relate to nucleic acid molecules encoding immunogenic proteins that have 98% homology to the nucleic acid coding sequences herein. Some embodiments relate to nucleic acid molecules encoding immunogenic proteins that have 99% homology to the nucleic acid coding sequences herein. In some embodiments, the nucleic acid molecules having coding sequences disclosed herein that are homologous to the coding sequences of the consensus proteins disclosed herein include a sequence encoding an IgE leader sequence linked to the 5' end of the coding sequence encoding the homologous protein sequence disclosed herein.

[0253] Some embodiments relate to nucleic acid sequences encoding proteins having a specified percentage identity to the full-length MAGEA4 consensus protein, immunogenic fragments of the MAGEA4 consensus protein, and immunogenic fragments of proteins having identity to the MAGEA4 consensus protein. Such nucleic acid molecules can be provided that encode immunogenic proteins having up to 80% identity to the full-length MAGEA4 consensus sequence, up to 85% identity to the full-length consensus sequence, up to 90% identity to the full-length MAGEA4 consensus sequence, up to 91% identity to the full-length MAGEA4 consensus sequence, up to 92% identity to the full-length MAGEA4 consensus sequence, up to 93% identity to the full-length MAGEA4 consensus sequence, up to 94% identity to the full-length MAGEA4 consensus sequence, up to 95% identity to the full-length MAGEA4 consensus sequence, up to 96% identity to the full-length MAGEA4 consensus sequence, up to 97% identity to the full-length MAGEA4 consensus sequence, up to 98% identity to the full-length MAGEA4 consensus sequence, and up to 99% identity to the full-length MAGEA4 consensus sequence. Likewise, nucleic acid sequences encoding immunogenic fragments set forth herein and immunogenic fragments of proteins having similar percent identities to the MAGEA4 proteins set forth herein as specified above are also provided.

[0254] In some embodiments, the nucleic acid sequence does not contain a coding sequence that encodes a leader sequence. In some embodiments, the nucleic acid sequence does not contain a coding sequence that encodes an IgE leader sequence.

[0255] Some embodiments relate to fragments of SEQ ID NO: 7. The fragments can be at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% of SEQ ID NO: 7. The fragments can have at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% homology to the fragments of SEQ ID NO: 7. The fragments can have at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to the fragments of SEQ ID NO: 7. In some embodiments, the fragment includes a sequence encoding a leader sequence, such as, for example, an immunoglobulin leader sequence, such as an IgE leader sequence. In some embodiments, the fragment does not contain a coding sequence encoding a leader sequence. In some embodiments, the fragment does not contain a coding sequence encoding a leader sequence, such as, for example, an IgE leader sequence.

[0256] In addition, the amino acid sequence of the consensus MAGEA4 protein is SEQ ID NO: 8. The amino acid sequence of the consensus MAGEA4 protein linked to the IgE leader sequence is SEQ ID NO: 8. The amino acid sequence of the consensus MAGEA4 protein linked to the IgE leader sequence can be linked to an HA tag.

[0257] Some embodiments relate to proteins homologous to SEQ ID NO: 8. Some embodiments relate to immunogenic proteins having 95% homology to the consensus protein sequence shown in SEQ ID NO: 8. Some embodiments relate to immunogenic proteins having 96% homology to the consensus protein sequence shown in SEQ ID NO: 8. Some embodiments relate to immunogenic proteins having 97% homology to the consensus protein sequence shown in SEQ ID NO: 8. Some embodiments relate to immunogenic proteins having 98% homology to the consensus protein sequence shown in SEQ ID NO: 8. Some embodiments relate to immunogenic proteins having 99% homology to the consensus protein sequence shown in SEQ ID NO: 8.

[0258] Some embodiments relate to proteins identical to SEQ ID NO: 8. Some embodiments relate to immunogenic proteins having an amino acid sequence that is 80% identical to the full-length consensus amino acid sequence as shown in SEQ ID NO: 8. Some embodiments relate to immunogenic proteins having an amino acid sequence that is 85% identical to the full-length consensus amino acid sequence as shown in SEQ ID NO: 8. Some embodiments relate to immunogenic proteins having an amino acid sequence that is 90% identical to the full-length consensus amino acid sequence as shown in SEQ ID NO: 8. Some embodiments relate to immunogenic proteins having an amino acid sequence that is 91% identical to the full-length consensus amino acid sequence as shown in SEQ ID NO: 8. Some embodiments relate to immunogenic proteins having an amino acid sequence that is 92% identical to the full-length consensus amino acid sequence as shown in SEQ ID NO: 8. Some embodiments relate to immunogenic proteins having an amino acid sequence that is 93% identical to the full-length consensus amino acid sequence as shown in SEQ ID NO: 8. Some embodiments relate to immunogenic proteins having an amino acid sequence that is 94% identical to the full-length consensus amino acid sequence as shown in SEQ ID NO: 8. Some embodiments relate to immunogenic proteins having an amino acid sequence that is 95% identical to the full-length consensus amino acid sequence as shown in SEQ ID NO: 8. Some embodiments relate to immunogenic proteins having an amino acid sequence that is 96% identical to the full-length consensus amino acid sequence as shown in SEQ ID NO: 8. Some embodiments relate to immunogenic proteins having an amino acid sequence that is 97% identical to the full-length consensus amino acid sequence as shown in SEQ ID NO: 8. Some embodiments relate to immunogenic proteins having an amino acid sequence that is 98% identical to the full-length consensus amino acid sequence as shown in SEQ ID NO: 8. Some embodiments relate to immunogenic proteins having an amino acid sequence that is 99% identical to the full-length consensus amino acid sequence as shown in SEQ ID NO: 8.

[0259] In some embodiments, the protein does not contain a leader sequence. In some embodiments, the protein does not contain an IgE leader sequence. A fragment of the consensus protein can comprise at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% of the consensus protein. An immunogenic fragment of SEQ ID NO: 8 can be provided. The immunogenic fragment may comprise at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% of SEQ ID NO: 8. In some embodiments, the fragment includes a leader sequence, such as, for example, an immunoglobulin leader sequence, such as an IgE leader sequence. In some embodiments, the fragment does not contain a leader sequence. In some embodiments, the fragment does not contain a leader sequence, such as, for example, an IgE leader sequence.

[0260] Immunogenic fragments of proteins having amino acid sequences homologous to immunogenic fragments of SEQ ID NO: 8 can be provided. Such immunogenic fragments can comprise at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% of a protein having 95% or greater homology to SEQ ID NO: 8. Some embodiments relate to immunogenic fragments having 96% homology to immunogenic fragments of the consensus protein sequences herein. Some embodiments relate to immunogenic fragments having 97% homology to immunogenic fragments of the consensus protein sequences herein. Some embodiments relate to immunogenic fragments having 98% homology to immunogenic fragments of the consensus protein sequences herein. Some embodiments relate to immunogenic fragments having 99% homology to immunogenic fragments of the consensus protein sequences herein. In some embodiments, the fragments include a leader sequence, such as, for example, an immunoglobulin leader sequence, such as an IgE leader sequence. In some embodiments, the fragments do not contain a leader sequence. In some embodiments, the fragments do not contain a leader sequence, such as, for example, an IgE leader sequence.

[0261] Immunogenic fragments of proteins having an amino acid sequence identical to an immunogenic fragment of SEQ ID NO: 8 can be provided. Such immunogenic fragments can comprise at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% of a protein having 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to the amino acid sequence shown in SEQ ID NO: 8. In some embodiments, the fragments include a leader sequence, such as, for example, an immunoglobulin leader sequence, such as an IgE leader sequence. In some embodiments, the fragments do not contain a leader sequence. In some embodiments, the fragment does not contain a leader sequence, such as, for example, an IgE leader sequence.

[0262] As indicated herein with respect to linking a signal peptide or leader sequence to the N-terminus of a protein, the signal peptide / leader sequence replaces the protein's N-terminal methionine that is encoded by the start codon of the nucleic acid sequence encoding the protein without the signal peptide coding sequence.

[0263] (5) Growth hormone-releasing hormone (GHRH)

[0264] The immunogenic compositions of the present invention may comprise the cancer antigen growth hormone-releasing hormone (GHRH; also known as growth-hormone-releasing factor (GRF or GHRF) or somatokinin), a fragment thereof, or a variant thereof. GHRH is a 44 amino acid peptide hormone produced in the arcuate nucleus of the hypothalamus. GHRH is secreted by the hypothalamus and stimulates the release of growth hormone, a regulator of growth, metabolism, and body structure, from the pituitary gland. GHRH also stimulates the production of growth hormone. Antagonists of GHRH can inhibit the growth of a variety of cancers, such as osteosarcoma, glioblastoma, prostate cancer, kidney cancer, pancreatic cancer, colorectal cancer, and breast cancer. Therefore, GHRH may be an antigen associated with a variety of tumors.

[0265] GHRH antigen can induce antigen-specific T cells and / or high-titer antibody responses, thereby inducing or triggering an immune response to a cancer or tumor expressing the antigen or to the cancer or tumor reaction. In some embodiments, the immune response induced or triggered can be a cellular immune response, a humoral immune response, or both a cellular and humoral immune response. In some embodiments, the cellular immune response induced or triggered may include the induction or secretion of interferon-γ (IFN-γ) and / or tumor necrosis factor α (TNF-α). In other embodiments, the immune response induced or triggered can reduce or inhibit one or more immunosuppressive factors that promote the growth of tumors or cancers expressing the antigen, for example, but not limited to factors that downregulate MHC presentation, factors that upregulate antigen-specific regulatory T cells (Treg), PD-L1, FasL, cytokines such as IL-10 and TFG-β, tumor-associated macrophages, tumor-associated fibroblasts, soluble factors produced by immunosuppressive cells, CTLA-4, PD-1, MDSC, MCP-1, and immune checkpoint molecules, described in more detail below.

[0266] GHRH antigens may comprise protein epitopes that make them particularly effective as immunogens against which an anti-GHRH immune response can be induced. GHRH antigens may comprise full-length translation products, variants thereof, fragments thereof, or combinations thereof. GHRH antigens may comprise a consensus protein.

[0267] The nucleic acid sequence encoding the consensus GHRH antigen can be optimized for codon usage and corresponding RNA transcripts. The nucleic acid encoding the consensus GHRH antigen can be codons and RNA optimized for expression. In some embodiments, the nucleic acid sequence encoding the consensus GHRH antigen can include a Kozak sequence (e.g., GCC ACC) to enhance the efficiency of translation. The nucleic acid encoding the consensus GHRH antigen can include multiple stop codons (e.g., TGA TGA) to enhance the efficiency of translation termination.

[0268] The nucleic acid encoding the consensus GHRH antigen may also encode an immunoglobulin E (IgE) leader sequence. The nucleic acid encoding the consensus GHRH antigen may further encode an IgE leader sequence such that the amino acid sequence of the IgE leader sequence is linked to the amino acid sequence of the consensus GHRH antigen via a peptide bond. The nucleic acid encoding the consensus GHRH antigen may also include a nucleotide sequence encoding the IgE leader sequence. In some embodiments, the nucleic acid encoding the consensus GHRH antigen is devoid of or does not contain a nucleotide sequence encoding the IgE leader sequence.

[0269] The consensus GHRH antigen can be the nucleic acid sequence of SEQ ID NO:9, which encodes the amino acid sequence of SEQ ID NO:10. SEQ ID NO:9 encodes the consensus GHRH protein linked to an IgE leader sequence. The consensus GHRH protein can be linked to an IgE leader sequence and an HA tag. In other embodiments, the consensus GHRH protein may not contain or be linked to an IgE leader sequence and / or an HA tag.

[0270] In some embodiments, the consensus GHRH antigen can be a nucleic acid sequence that is at least about 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical over the entire length of the nucleic acid sequence shown in SEQ ID NO: 9. In other embodiments, the consensus GHRH antigen can be a nucleic acid sequence that encodes an amino acid sequence that is at least about 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical over the entire length of the amino acid sequence shown in SEQ ID NO: 10. The consensus GHRH antigen can be an amino acid sequence that is at least about 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical over the entire length of the amino acid sequence shown in SEQ ID NO: 10.

[0271] Some embodiments relate to nucleic acid sequences encoding proteins homologous to the GHRH consensus protein, immunogenic fragments of the GHRH consensus protein, and immunogenic fragments of homologous proteins. Such nucleic acid molecules encoding immunogenic proteins having up to 95% homology to the consensus sequence, up to 96% homology to the consensus sequence, up to 97% homology to the consensus sequence, up to 98% homology to the consensus sequence, and up to 99% homology to the consensus sequence can be provided. Similarly, nucleic acid sequences encoding immunogenic fragments as set forth herein and immunogenic fragments of proteins homologous to the proteins set forth herein are also provided.

[0272] Some embodiments relate to nucleic acid molecules encoding immunogenic proteins that have 95% homology to the nucleic acid coding sequences herein. Some embodiments relate to nucleic acid molecules encoding immunogenic proteins that have 96% homology to the nucleic acid coding sequences herein. Some embodiments relate to nucleic acid molecules encoding immunogenic proteins that have 97% homology to the nucleic acid coding sequences herein. Some embodiments relate to nucleic acid molecules encoding immunogenic proteins that have 98% homology to the nucleic acid coding sequences herein. Some embodiments relate to nucleic acid molecules encoding immunogenic proteins that have 99% homology to the nucleic acid coding sequences herein. In some embodiments, the nucleic acid molecules having coding sequences disclosed herein that are homologous to the coding sequences of the consensus proteins disclosed herein include a sequence encoding an IgE leader sequence linked to the 5' end of the coding sequence encoding the homologous protein sequence disclosed herein.

[0273] Some embodiments relate to nucleic acid sequences encoding proteins having a specific percentage identity to a full-length GHRH consensus protein, immunogenic fragments of a GHRH consensus protein, and immunogenic fragments of a protein having identity to a GHRH consensus protein. Such nucleic acid molecules can be provided that encode immunogenic proteins having up to 80% identity to the full-length GHRH consensus sequence, up to 85% identity to the full-length consensus sequence, up to 90% identity to the full-length GHRH consensus sequence, up to 91% identity to the full-length GHRH consensus sequence, up to 92% identity to the full-length GHRH consensus sequence, up to 93% identity to the full-length GHRH consensus sequence, up to 94% identity to the full-length GHRH consensus sequence, up to 95% identity to the full-length GHRH consensus sequence, up to 96% identity to the full-length GHRH consensus sequence, up to 97% identity to the full-length GHRH consensus sequence, up to 98% identity to the full-length GHRH consensus sequence, and up to 99% identity to the full-length GHRH consensus sequence. Similarly, nucleic acid sequences encoding immunogenic fragments as set forth herein and immunogenic fragments of proteins having similar percentages of identity to the GHRH proteins set forth herein as specified above are also provided.

[0274] In some embodiments, the nucleic acid sequence does not contain a coding sequence that encodes a leader sequence. In some embodiments, the nucleic acid sequence does not contain a coding sequence that encodes an IgE leader sequence.

[0275] Some embodiments relate to fragments of SEQ ID NO: 9. The fragments can be at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% of SEQ ID NO: 9. The fragments can have at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% homology to the fragments of SEQ ID NO: 9. The fragments can have at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to the fragments of SEQ ID NO: 9. In some embodiments, the fragment includes a sequence encoding a leader sequence, such as, for example, an immunoglobulin leader sequence, such as an IgE leader sequence. In some embodiments, the fragment does not contain a coding sequence encoding a leader sequence. In some embodiments, the fragment does not contain a coding sequence encoding a leader sequence, such as, for example, an IgE leader sequence.

[0276] In addition, the amino acid sequence of the consensus GHRH protein is SEQ ID NO: 10. The amino acid sequence of the consensus GHRH protein linked to the IgE leader sequence is SEQ ID NO: 10. The amino acid sequence of the consensus GHRH protein linked to the IgE leader sequence can be linked to an HA tag.

[0277] Some embodiments relate to proteins homologous to SEQ ID NO: 10. Some embodiments relate to immunogenic proteins having 95% homology to the consensus protein sequence shown in SEQ ID NO: 10. Some embodiments relate to immunogenic proteins having 96% homology to the consensus protein sequence shown in SEQ ID NO: 10. Some embodiments relate to immunogenic proteins having 97% homology to the consensus protein sequence shown in SEQ ID NO: 10. Some embodiments relate to immunogenic proteins having 98% homology to the consensus protein sequence shown in SEQ ID NO: 10. Some embodiments relate to immunogenic proteins having 99% homology to the consensus protein sequence shown in SEQ ID NO: 10.

[0278] Some embodiments relate to proteins identical to SEQ ID NO: 10. Some embodiments relate to immunogenic proteins having an amino acid sequence that is 80% identical to the full-length consensus amino acid sequence as shown in SEQ ID NO: 10. Some embodiments relate to immunogenic proteins having an amino acid sequence that is 85% identical to the full-length consensus amino acid sequence as shown in SEQ ID NO: 10. Some embodiments relate to immunogenic proteins having an amino acid sequence that is 90% identical to the full-length consensus amino acid sequence as shown in SEQ ID NO: 10. Some embodiments relate to immunogenic proteins having an amino acid sequence that is 91% identical to the full-length consensus amino acid sequence as shown in SEQ ID NO: 10. Some embodiments relate to immunogenic proteins having an amino acid sequence that is 92% identical to the full-length consensus amino acid sequence as shown in SEQ ID NO: 10. Some embodiments relate to immunogenic proteins having an amino acid sequence that is 93% identical to the full-length consensus amino acid sequence as shown in SEQ ID NO: 10. Some embodiments relate to immunogenic proteins having an amino acid sequence that is 94% identical to the full-length consensus amino acid sequence as shown in SEQ ID NO: 10. Some embodiments relate to immunogenic proteins having an amino acid sequence that is 95% identical to the full-length consensus amino acid sequence as shown in SEQ ID NO: 10. Some embodiments relate to immunogenic proteins having an amino acid sequence that is 96% identical to the full-length consensus amino acid sequence as shown in SEQ ID NO: 10. Some embodiments relate to immunogenic proteins having an amino acid sequence that is 97% identical to the full-length consensus amino acid sequence as shown in SEQ ID NO: 10. Some embodiments relate to immunogenic proteins having an amino acid sequence that is 98% identical to the full-length consensus amino acid sequence as shown in SEQ ID NO: 10. Some embodiments relate to immunogenic proteins having an amino acid sequence that is 99% identical to the full-length consensus amino acid sequence as shown in SEQ ID NO: 10.

[0279] In some embodiments, the protein does not contain a leader sequence. In some embodiments, the protein does not contain an IgE leader sequence. A fragment of the consensus protein can comprise at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% of the consensus protein. An immunogenic fragment of SEQ ID NO: 10 can be provided. The immunogenic fragment may comprise at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% of SEQ ID NO: 10. In some embodiments, the fragment includes a leader sequence, such as, for example, an immunoglobulin leader sequence, such as an IgE leader sequence. In some embodiments, the fragment does not contain a leader sequence. In some embodiments, the fragment does not contain a leader sequence, such as, for example, an IgE leader sequence.

[0280] Immunogenic fragments of proteins having amino acid sequences homologous to immunogenic fragments of SEQ ID NO: 10 can be provided. Such immunogenic fragments can comprise at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% of a protein having 95% or greater homology to SEQ ID NO: 10. Some embodiments relate to immunogenic fragments having 96% homology to immunogenic fragments of the consensus protein sequences herein. Some embodiments relate to immunogenic fragments having 97% homology to immunogenic fragments of the consensus protein sequences herein. Some embodiments relate to immunogenic fragments having 98% homology to an immunogenic fragment of a consensus protein sequence herein. Some embodiments relate to immunogenic fragments having 99% homology to an immunogenic fragment of a consensus protein sequence herein. In some embodiments, the fragment includes a leader sequence, such as, for example, an immunoglobulin leader sequence, such as an IgE leader sequence. In some embodiments, the fragment does not contain a leader sequence. In some embodiments, the fragment does not contain a leader sequence, such as, for example, an IgE leader sequence.

[0281] Immunogenic fragments of proteins having an amino acid sequence identical to an immunogenic fragment of SEQ ID NO: 10 can be provided. Such immunogenic fragments can comprise at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% of a protein having 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to the amino acid sequence shown in SEQ ID NO: 10. In some embodiments, the fragments include a leader sequence, such as, for example, an immunoglobulin leader sequence, such as an IgE leader sequence. In some embodiments, the fragments do not contain a leader sequence. In some embodiments, the fragment does not contain a leader sequence, such as, for example, an IgE leader sequence.

[0282] As indicated herein with respect to linking a signal peptide or leader sequence to the N-terminus of a protein, the signal peptide / leader sequence replaces the protein's N-terminal methionine that is encoded by the start codon of the nucleic acid sequence encoding the protein without the signal peptide coding sequence.

[0283] (6)MART-1 / Melan-A

[0284] The immunogenic compositions of the present invention may comprise the cancer antigen MART-1 (also known as Melan-A), a fragment thereof or a variant thereof. MART-1, encoded by the MLANA gene, is a 118-amino acid protein containing a single transmembrane domain and is expressed in most melanoma cells. MART-1 forms a complex with structural proteins and affects the transport and processing required for its expression, stability, melanosome structure and maturation. Therefore, MART-1 is necessary for regulating pigmentation in mammals. Defects in melanosome maturation are associated with susceptibility to cancer. MART-1 can be expressed in many types of cancer (including but not limited to melanoma).

[0285] Melan-A, also known as melanoma antigen recognized by T cells (MART-1), is a melanocyte differentiation antigen and is found in normal skin, retina and melanocytes. Melan-a can be associated with the endoplasmic reticulum and melanosomes. Melan-A can be recognized as an antigen on melanoma cells by cytotoxic T cells, but can also be associated with other tumors such as clear cell sarcoma and melanin-type neurofibroma with melanocyte origin or differentiation (i.e., cells have melanosomes). Therefore, Melan-A can be an antigen associated with a variety of tumors derived from cells with melanosomes.

[0286] Melan-A antigen can induce antigen-specific T cells and / or high-titer antibody responses, thereby inducing or triggering an immune response to a cancer or tumor expressing the antigen or to the cancer or tumor reaction. In some embodiments, the immune response induced or triggered can be a cellular immune response, a humoral immune response, or both a cellular and humoral immune response. In some embodiments, the cellular immune response induced or triggered may include the induction or secretion of interferon-γ (IFN-γ) and / or tumor necrosis factor α (TNF-α). In other embodiments, the immune response induced or triggered can reduce or inhibit one or more immunosuppressive factors that promote the growth of tumors or cancers expressing the antigen, for example, but not limited to factors that downregulate MHC presentation, factors that upregulate antigen-specific regulatory T cells (Treg), PD-L1, FasL, cytokines such as IL-10 and TFG-β, tumor-associated macrophages, tumor-associated fibroblasts, soluble factors produced by immunosuppressive cells, CTLA-4, PD-1, MDSC, MCP-1, and immune checkpoint molecules, described in more detail below.

[0287] Melan-A antigens may comprise protein epitopes that make them particularly effective as immunogens against which an anti-Melan-A immune response can be induced. Melan-A antigens may comprise full-length translation products, variants thereof, fragments thereof, or combinations thereof. Melan-A antigens may comprise a consensus protein.

[0288] The nucleic acid sequence encoding the consensus Melan-A antigen can be optimized for codon usage and corresponding RNA transcripts. The nucleic acid encoding the consensus Melan-A antigen can be codons and RNA optimized for expression. In some embodiments, the nucleic acid sequence encoding the consensus Melan-A antigen can include a Kozak sequence (e.g., GCC ACC) to enhance the efficiency of translation. The nucleic acid encoding the consensus Melan-A antigen can include multiple stop codons (e.g., TGA TGA) to enhance the efficiency of translation termination.

[0289] The nucleic acid encoding the consensus Melan-A antigen may also encode an immunoglobulin E (IgE) leader sequence. The nucleic acid encoding the consensus Melan-A antigen may further encode an IgE leader sequence such that the amino acid sequence of the IgE leader sequence is linked to the amino acid sequence of the consensus Melan-A antigen via a peptide bond. The nucleic acid encoding the consensus Melan-A antigen may also include a nucleotide sequence encoding the IgE leader sequence. In some embodiments, the nucleic acid encoding the consensus Melan-A antigen does not have or contains a nucleotide sequence encoding the IgE leader sequence.

[0290] The consensus Melan-A antigen can be the nucleic acid sequence SEQ ID NO: 11, which encodes the amino acid sequence SEQ ID NO: 12. SEQ ID NO: 11 encodes the consensus Melan-A protein linked to an IgE leader sequence. The consensus Melan-A protein can be linked to an IgE leader sequence and an HA tag. In other embodiments, the consensus Melan-A protein may not contain or be linked to an IgE leader sequence and / or an HA tag.

[0291] In some embodiments, the consensus Melan-A antigen can be a nucleic acid sequence that is at least about 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical over the entire length of the nucleic acid sequence shown in SEQ ID NO: 11. In other embodiments, the consensus Melan-A antigen can be a nucleic acid sequence that encodes an amino acid sequence that is at least about 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical over the entire length of the amino acid sequence shown in SEQ ID NO: 12. The consensus Melan-A antigen can be an amino acid sequence that is at least about 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical over the entire length of the amino acid sequence shown in SEQ ID NO:12.

[0292] Some embodiments relate to nucleic acid sequences encoding proteins homologous to the Melan-A consensus protein, immunogenic fragments of the Melan-A consensus protein, and immunogenic fragments of homologous proteins. Such nucleic acid molecules encoding immunogenic proteins having up to 95% homology to the consensus sequence, up to 96% homology to the consensus sequence, up to 97% homology to the consensus sequence, up to 98% homology to the consensus sequence, and up to 99% homology to the consensus sequence can be provided. Similarly, nucleic acid sequences encoding immunogenic fragments as set forth herein and immunogenic fragments of proteins homologous to the proteins set forth herein are also provided.

[0293] Some embodiments relate to nucleic acid molecules encoding immunogenic proteins that have 95% homology to the nucleic acid coding sequences herein. Some embodiments relate to nucleic acid molecules encoding immunogenic proteins that have 96% homology to the nucleic acid coding sequences herein. Some embodiments relate to nucleic acid molecules encoding immunogenic proteins that have 97% homology to the nucleic acid coding sequences herein. Some embodiments relate to nucleic acid molecules encoding immunogenic proteins that have 98% homology to the nucleic acid coding sequences herein. Some embodiments relate to nucleic acid molecules encoding immunogenic proteins that have 99% homology to the nucleic acid coding sequences herein. In some embodiments, the nucleic acid molecules having coding sequences disclosed herein that are homologous to the coding sequences of the consensus proteins disclosed herein include a sequence encoding an IgE leader sequence linked to the 5' end of the coding sequence encoding the homologous protein sequence disclosed herein.

[0294] Some embodiments relate to nucleic acid sequences encoding proteins having a specific percent identity to a full-length Melan-A consensus protein, immunogenic fragments of a Melan-A consensus protein, and immunogenic fragments of proteins having identity to a Melan-A consensus protein. Such nucleic acid molecules can be provided that encode immunogenic proteins having up to 80% identity to the full-length Melan-A consensus sequence, up to 85% identity to the full-length Melan-A consensus sequence, up to 90% identity to the full-length Melan-A consensus sequence, up to 91% identity to the full-length Melan-A consensus sequence, up to 92% identity to the full-length Melan-A consensus sequence, up to 93% identity to the full-length Melan-A consensus sequence, up to 94% identity to the full-length Melan-A consensus sequence, up to 95% identity to the full-length Melan-A consensus sequence, up to 96% identity to the full-length Melan-A consensus sequence, up to 97% identity to the full-length Melan-A consensus sequence, up to 98% identity to the full-length Melan-A consensus sequence, and up to 99% identity to the full-length Melan-A consensus sequence. Likewise, nucleic acid sequences encoding immunogenic fragments set forth herein and immunogenic fragments of proteins having similar percent identities to the Melan-A proteins set forth herein as specified above are also provided.

[0295] In some embodiments, the nucleic acid sequence does not contain a coding sequence that encodes a leader sequence. In some embodiments, the nucleic acid sequence does not contain a coding sequence that encodes an IgE leader sequence.

[0296] Some embodiments relate to fragments of SEQ ID NO: 11. The fragments can be at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% of SEQ ID NO: 11. The fragments can be at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% homologous to a fragment of SEQ ID NO: 11. The fragment may be at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to a fragment of SEQ ID NO: 11. In some embodiments, the fragment includes a sequence encoding a leader sequence, such as, for example, an immunoglobulin leader sequence, such as an IgE leader sequence. In some embodiments, the fragment does not contain a coding sequence encoding a leader sequence. In some embodiments, the fragment does not contain a coding sequence encoding a leader sequence, such as, for example, an IgE leader sequence.

[0297] In addition, the amino acid sequence of the consensus Melan-A protein is SEQ ID NO: 12. The amino acid sequence of the consensus Melan-A protein linked to the IgE leader sequence is SEQ ID NO: 12. The amino acid sequence of the consensus Melan-A protein linked to the IgE leader sequence can be linked to an HA tag.

[0298] Some embodiments relate to proteins homologous to SEQ ID NO: 12. Some embodiments relate to immunogenic proteins having 95% homology to the consensus protein sequence shown in SEQ ID NO: 12. Some embodiments relate to immunogenic proteins having 96% homology to the consensus protein sequence shown in SEQ ID NO: 12. Some embodiments relate to immunogenic proteins having 97% homology to the consensus protein sequence shown in SEQ ID NO: 12. Some embodiments relate to immunogenic proteins having 98% homology to the consensus protein sequence shown in SEQ ID NO: 12. Some embodiments relate to immunogenic proteins having 99% homology to the consensus protein sequence shown in SEQ ID NO: 12.

[0299] Some embodiments relate to a protein identical to SEQ ID NO: 12. Some embodiments relate to an immunogenic protein having an amino acid sequence that is 80% identical to the full-length consensus amino acid sequence as shown in SEQ ID NO: 12. Some embodiments relate to an immunogenic protein having an amino acid sequence that is 85% identical to the full-length consensus amino acid sequence as shown in SEQ ID NO: 12. Some embodiments relate to an immunogenic protein having an amino acid sequence that is 90% identical to the full-length consensus amino acid sequence as shown in SEQ ID NO: 12. Some embodiments relate to an immunogenic protein having an amino acid sequence that is 91% identical to the full-length consensus amino acid sequence as shown in SEQ ID NO: 12. Some embodiments relate to an immunogenic protein having an amino acid sequence that is 92% identical to the full-length consensus amino acid sequence as shown in SEQ ID NO: 12. Some embodiments relate to an immunogenic protein having an amino acid sequence that is 93% identical to the full-length consensus amino acid sequence as shown in SEQ ID NO: 12. Some embodiments relate to immunogenic proteins having an amino acid sequence that is 94% identical to the full-length consensus amino acid sequence as shown in SEQ ID NO: 12. Some embodiments relate to immunogenic proteins having an amino acid sequence that is 95% identical to the full-length consensus amino acid sequence as shown in SEQ ID NO: 12. Some embodiments relate to immunogenic proteins having an amino acid sequence that is 96% identical to the full-length consensus amino acid sequence as shown in SEQ ID NO: 12. Some embodiments relate to immunogenic proteins having an amino acid sequence that is 97% identical to the full-length consensus amino acid sequence as shown in SEQ ID NO: 12. Some embodiments relate to immunogenic proteins having an amino acid sequence that is 98% identical to the full-length consensus amino acid sequence as shown in SEQ ID NO: 12. Some embodiments relate to immunogenic proteins having an amino acid sequence that is 99% identical to the full-length consensus amino acid sequence as shown in SEQ ID NO: 12.

[0300] In some embodiments, the protein does not contain a leader sequence. In some embodiments, the protein does not contain an IgE leader sequence. A fragment of the consensus protein can comprise at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% of the consensus protein. An immunogenic fragment of SEQ ID NO: 12 can be provided. The immunogenic fragment may comprise at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% of SEQ ID NO: 12. In some embodiments, the fragment includes a leader sequence, such as, for example, an immunoglobulin leader sequence, such as an IgE leader sequence. In some embodiments, the fragment does not contain a leader sequence. In some embodiments, the fragment does not contain a leader sequence, such as, for example, an IgE leader sequence.

[0301] Immunogenic fragments of proteins having amino acid sequences homologous to immunogenic fragments of SEQ ID NO: 12 can be provided. Such immunogenic fragments can comprise at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% of a protein having 95% or greater homology to SEQ ID NO: 12. Some embodiments relate to immunogenic fragments having 96% homology to immunogenic fragments of the consensus protein sequences herein. Some embodiments relate to immunogenic fragments having 97% homology to immunogenic fragments of the consensus protein sequences herein. Some embodiments relate to immunogenic fragments having 98% homology to an immunogenic fragment of a consensus protein sequence herein. Some embodiments relate to immunogenic fragments having 99% homology to an immunogenic fragment of a consensus protein sequence herein. In some embodiments, the fragment includes a leader sequence, such as, for example, an immunoglobulin leader sequence, such as an IgE leader sequence. In some embodiments, the fragment does not contain a leader sequence. In some embodiments, the fragment does not contain a leader sequence, such as, for example, an IgE leader sequence.

[0302] Immunogenic fragments of proteins having an amino acid sequence identical to an immunogenic fragment of SEQ ID NO: 12 can be provided. Such immunogenic fragments can comprise at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% of a protein having 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to the amino acid sequence shown in SEQ ID NO: 12. In some embodiments, the fragments include a leader sequence, such as, for example, an immunoglobulin leader sequence, such as an IgE leader sequence. In some embodiments, the fragments do not contain a leader sequence. In some embodiments, the fragment does not contain a leader sequence, such as, for example, an IgE leader sequence.

[0303] As indicated herein with respect to linking a signal peptide or leader sequence to the N-terminus of a protein, the signal peptide / leader sequence replaces the protein's N-terminal methionine that is encoded by the start codon of the nucleic acid sequence encoding the protein without the signal peptide coding sequence.

[0304] (7)NY-ESO-1

[0305] The immunogenic compositions of the present invention may comprise the cancer antigen New York Esophageal Cancer-1 (NY-ESO-1; also known as CTAG1), fragments thereof, or variants thereof. NY-ESO-1, encoded by the CTAG1B gene, is a 180-amino acid protein with a glycine-rich N-terminal region and an extremely hydrophobic C-terminal region. NY-ESO-1 has limited expression in normal tissues but is frequently found in cancer. NY-ESO-1 is expressed in a variety of cancers, including, but not limited to, bladder, colorectal, esophageal, gastric, liver, head and neck cancers, melanoma, non-small cell lung cancer, ovarian cancer, pancreatic cancer, synovial cancer, and prostate cancer.

[0306] Cancer-testis antigen (NY-ESO-1) is expressed in the testes and ovaries. NY-ESO-1 is associated with various cancers and can induce humoral immune responses. Subjects with cancer or tumors can develop immunogenicity against NY-ESO-1. Therefore, NY-ESO-1 may be an antigen associated with various tumors.

[0307] The NY-ESO-1 antigen can induce antigen-specific T cell and / or high-titer antibody responses, thereby inducing or eliciting an immune response to or reactive with a cancer or tumor expressing the antigen. In some embodiments, the induced or elicited immune response can be a cellular immune response, a humoral immune response, or both a cellular and humoral immune response. In some embodiments, the induced or elicited cellular immune response can include the induction or secretion of interferon-γ (IFN-γ) and / or tumor necrosis factor α (TNF-α). In other embodiments, the induced or elicited immune response can reduce or inhibit one or more immunosuppressive factors that promote the growth of tumors or cancers expressing the antigen, such as, but not limited to, factors that downregulate MHC presentation, factors that upregulate antigen-specific regulatory T cells (Tregs), PD-L1, FasL, cytokines such as IL-10 and TFG-β, tumor-associated macrophages, tumor-associated fibroblasts, soluble factors produced by immunosuppressive cells, CTLA-4, PD-1, MDSC, MCP-1, and immune checkpoint molecules, which are described in more detail below.

[0308] The NY-ESO-1 antigen can enhance the cellular immune response of a subject administered with the NY-ESO-1 antigen by about 50-fold to about 6,000-fold, about 50-fold to about 5,500-fold, about 50-fold to about 5,000-fold, about 50-fold to about 4,500-fold, about 100-fold to about 6,000-fold, about 150-fold to about 6,000-fold, about 200-fold to about 6,000-fold, about 250-fold to about 6,000-fold, or about 300-fold to about 6,000-fold, compared to the cellular immune response of a subject not administered with the NY-ESO-1 antigen. In some embodiments, the NY-ESO-1 antigen can enhance the cellular immune response of a subject administered the NY-ESO-1 antigen by about 50-fold, 100-fold, 150-fold, 200-fold, 250-fold, 300-fold, 350-fold, 400-fold, 450-fold, 500-fold, 550-fold, 600-fold, 650-fold, 700-fold, 750-fold, 800-fold, 850-fold, 900-fold, 950-fold, 1000-fold, 1100-fold, 1200-fold, 1300-fold, 1400-fold, 1500-fold, 1600-fold, 1700-fold, 1800-fold, 1900-fold, 2000-fold, 2100-fold, 2200-fold, 2300-fold, 2400-fold, 2500-fold, 2600-fold, 2700-fold, 2800-fold, 2900-fold, 3000-fold, 3100-fold, 3200-fold, 3300-fold, 3400-fold, 3500-fold, 3600-fold, 3700-fold, 3800-fold, 3900-fold, 4000-fold, 4100-fold, 4200-fold, 4300-fold, 4400-fold 00 times, 2200 times, 2300 times, 2400 times, 2500 times, 2600 times, 2700 times, 2800 times, 2900 times, 3000 times, 3100 times, 3200 times, 3300 times, 3400 times, 3500 times, 3600 times, 3700 times, 3800 times, 3900 times, 4000 times, 4100 times, 4200 times, 4300 times, 4400 times, 4500 times, 4600 times, 4700 times, 4800 times, 4900 times, 5000 times, 5100 times, 5200 times, 5300 times, 5400 times, 5500 times, 5600 times, 5700 times, 5800 times, 5900 times or 6000 times.

[0309] The NY-ESO-1 antigen can increase interferon gamma (IFN-γ) levels in a subject administered with the NY-ESO-1 antigen by about 50-fold to about 6,000-fold, about 50-fold to about 5,500-fold, about 50-fold to about 5,000-fold, about 50-fold to about 4,500-fold, about 100-fold to about 6,000-fold, about 150-fold to about 6,000-fold, about 200-fold to about 6,000-fold, about 250-fold to about 6,000-fold, or about 300-fold to about 6,000-fold, compared to IFN-γ levels in a subject not administered with the NY-ESO-1 antigen. In some embodiments, the NY-ESO-1 antigen can increase IFN-γ levels in a subject administered with the NY-ESO-1 antigen by about 50-fold, 100-fold, 150-fold, 200-fold, 250-fold, 300-fold, 350-fold, 400-fold, 450-fold, 500-fold, 550-fold, 600-fold, 650-fold, 700-fold, 750-fold, 800-fold, 850-fold, 900-fold, 950-fold, 1000-fold, 1100-fold, 1200-fold, 1300-fold, 1400-fold, 1500-fold, 1600-fold, 1700-fold, 1800-fold, 1900-fold, 2000-fold, 2100-fold, 2200-fold, 2300-fold, 2400-fold, 2500-fold, 2600-fold, 2700-fold, 2800-fold, 2900-fold, 3000-fold, 3100-fold, 3200-fold, 3300-fold, 3400-fold, 3500-fold, 3600-fold, 3700-fold, 3800-fold, 3900-fold, 4000-fold, 4100-fold, 4200-fold, 4300-fold, 4400-fold 100 times, 2200 times, 2300 times, 2400 times, 2500 times, 2600 times, 2700 times, 2800 times, 2900 times, 3000 times, 3100 times, 3200 times, 3300 times, 3400 times, 3500 times, 3600 times, 3700 times, 3800 times, 3900 times, 4000 times, 4100 times, 4200 times, 4300 times, 4400 times, 4500 times, 4600 times, 4700 times, 4800 times, 4900 times, 5000 times, 5100 times, 5200 times, 5300 times, 5400 times, 5500 times, 5600 times, 5700 times, 5800 times, 5900 times or 6000 times.

[0310] NY-ESO-1 antigens may comprise protein epitopes that make them particularly effective as immunogens against which an anti-NY-ESO-1 immune response can be induced. NY-ESO-1 antigens may comprise full-length translation products, variants thereof, fragments thereof, or combinations thereof. NY-ESO-1 antigens may comprise consensus proteins.

[0311] Nucleic acid sequences encoding the consensus NY-ESO-1 antigen can be optimized for codon usage and corresponding RNA transcripts. Nucleic acids encoding the consensus NY-ESO-1 antigen can be codon- and RNA-optimized for expression. In some embodiments, the nucleic acid sequence encoding the consensus NY-ESO-1 antigen can include a Kozak sequence (e.g., GCC ACC) to enhance translation efficiency. Nucleic acids encoding the consensus NY-ESO-1 antigen can include multiple stop codons (e.g., TGA TGA) to enhance translation termination efficiency.

[0312] The nucleic acid encoding the consensus NY-ESO-1 antigen may also encode an immunoglobulin E (IgE) leader sequence. The nucleic acid encoding the consensus NY-ESO-1 antigen may further encode an IgE leader sequence such that the amino acid sequence of the IgE leader sequence is linked to the amino acid sequence of the consensus NY-ESO-1 antigen via a peptide bond. The nucleic acid encoding the consensus NY-ESO-1 antigen may also include a nucleotide sequence encoding the IgE leader sequence. In some embodiments, the nucleic acid encoding the consensus NY-ESO-1 antigen lacks or contains a nucleotide sequence encoding the IgE leader sequence.

[0313] The consensus NY-ESO-1 antigen can be the nucleic acid sequence of SEQ ID NO: 13, which encodes the amino acid sequence of SEQ ID NO: 14. SEQ ID NO: 13 encodes the consensus NY-ESO-1 protein linked to an IgE leader sequence. The consensus NY-ESO-1 protein can be linked to an IgE leader sequence and an HA tag. In other embodiments, the consensus NY-ESO-1 protein may not contain or be linked to an IgE leader sequence and / or an HA tag.

[0314] In some embodiments, the consensus NY-ESO-1 antigen can be a nucleic acid sequence that is at least about 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical over the entire length of the nucleic acid sequence shown in SEQ ID NO: 13. In other embodiments, the consensus NY-ESO-1 antigen can be a nucleic acid sequence that encodes an amino acid sequence that is at least about 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical over the entire length of the amino acid sequence shown in SEQ ID NO: 14. The consensus NY-ESO-1 antigen can be an amino acid sequence that is at least about 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical over the entire length of the amino acid sequence shown in SEQ ID NO: 14.

[0315] Some embodiments relate to nucleic acid sequences encoding proteins homologous to the NY-ESO-1 consensus protein, immunogenic fragments of the NY-ESO-1 consensus protein, and immunogenic fragments of homologous proteins. Such nucleic acid molecules can be provided that encode immunogenic proteins having up to 95% homology to the consensus sequence, up to 96% homology to the consensus sequence, up to 97% homology to the consensus sequence, up to 98% homology to the consensus sequence, and up to 99% homology to the consensus sequence. Similarly, nucleic acid sequences encoding immunogenic fragments as described herein and immunogenic fragments of proteins homologous to the proteins described herein are also provided.

[0316] Some embodiments relate to nucleic acid molecules encoding immunogenic proteins that have 95% homology to the nucleic acid coding sequences herein. Some embodiments relate to nucleic acid molecules encoding immunogenic proteins that have 96% homology to the nucleic acid coding sequences herein. Some embodiments relate to nucleic acid molecules encoding immunogenic proteins that have 97% homology to the nucleic acid coding sequences herein. Some embodiments relate to nucleic acid molecules encoding immunogenic proteins that have 98% homology to the nucleic acid coding sequences herein. Some embodiments relate to nucleic acid molecules encoding immunogenic proteins that have 99% homology to the nucleic acid coding sequences herein. In some embodiments, the nucleic acid molecules having coding sequences disclosed herein that are homologous to the coding sequences of the consensus proteins disclosed herein include a sequence encoding an IgE leader sequence linked to the 5' end of the coding sequence encoding the homologous protein sequence disclosed herein.

[0317] Some embodiments relate to nucleic acid sequences encoding proteins having a specific percentage identity to a full-length NY-ESO-1 consensus protein, immunogenic fragments of a NY-ESO-1 consensus protein, and immunogenic fragments of a protein identical to a NY-ESO-1 consensus protein. Such nucleic acid molecules can be provided that encode immunogenic proteins having up to 80% identity to the full-length NY-ESO-1 consensus sequence, up to 85% identity to the full-length consensus sequence, up to 90% identity to the full-length NY-ESO-1 consensus sequence, up to 91% identity to the full-length NY-ESO-1 consensus sequence, up to 92% identity to the full-length NY-ESO-1 consensus sequence, up to 93% identity to the full-length NY-ESO-1 consensus sequence, up to 94% identity to the full-length NY-ESO-1 consensus sequence, up to 95% identity to the full-length NY-ESO-1 consensus sequence, up to 96% identity to the full-length NY-ESO-1 consensus sequence, up to 97% identity to the full-length NY-ESO-1 consensus sequence, up to 98% identity to the full-length NY-ESO-1 consensus sequence, and up to 99% identity to the full-length NY-ESO-1 consensus sequence. Likewise, nucleic acid sequences encoding immunogenic fragments set forth herein and immunogenic fragments of proteins having similar percent identities to the NY-ESO-1 proteins set forth herein as specified above are also provided.

[0318] In some embodiments, the nucleic acid sequence does not contain a coding sequence that encodes a leader sequence. In some embodiments, the nucleic acid sequence does not contain a coding sequence that encodes an IgE leader sequence.

[0319] Some embodiments relate to fragments of SEQ ID NO: 13. The fragments can be at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% of SEQ ID NO: 13. The fragments can be at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% homologous to a fragment of SEQ ID NO: 13. The fragment may be at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to a fragment of SEQ ID NO: 13. In some embodiments, the fragment includes a sequence encoding a leader sequence, such as, for example, an immunoglobulin leader sequence, such as an IgE leader sequence. In some embodiments, the fragment does not contain a coding sequence encoding a leader sequence. In some embodiments, the fragment does not contain a coding sequence encoding a leader sequence, such as, for example, an IgE leader sequence.

[0320] In addition, the amino acid sequence of the consensus NY-ESO-1 protein is SEQ ID NO: 14. The amino acid sequence of the consensus NY-ESO-1 protein linked to the IgE leader sequence is SEQ ID NO: 14. The amino acid sequence of the consensus NY-ESO-1 protein linked to the IgE leader sequence can be linked to an HA tag.

[0321] Some embodiments relate to proteins homologous to SEQ ID NO: 14. Some embodiments relate to immunogenic proteins having 95% homology to the consensus protein sequence shown in SEQ ID NO: 14. Some embodiments relate to immunogenic proteins having 96% homology to the consensus protein sequence shown in SEQ ID NO: 14. Some embodiments relate to immunogenic proteins having 97% homology to the consensus protein sequence shown in SEQ ID NO: 14. Some embodiments relate to immunogenic proteins having 98% homology to the consensus protein sequence shown in SEQ ID NO: 14. Some embodiments relate to immunogenic proteins having 99% homology to the consensus protein sequence shown in SEQ ID NO: 14.

[0322] Some embodiments relate to a protein identical to SEQ ID NO: 14. Some embodiments relate to an immunogenic protein having an amino acid sequence that is 80% identical to the full-length consensus amino acid sequence as shown in SEQ ID NO: 14. Some embodiments relate to an immunogenic protein having an amino acid sequence that is 85% identical to the full-length consensus amino acid sequence as shown in SEQ ID NO: 14. Some embodiments relate to an immunogenic protein having an amino acid sequence that is 90% identical to the full-length consensus amino acid sequence as shown in SEQ ID NO: 14. Some embodiments relate to an immunogenic protein having an amino acid sequence that is 91% identical to the full-length consensus amino acid sequence as shown in SEQ ID NO: 14. Some embodiments relate to an immunogenic protein having an amino acid sequence that is 92% identical to the full-length consensus amino acid sequence as shown in SEQ ID NO: 14. Some embodiments relate to an immunogenic protein having an amino acid sequence that is 93% identical to the full-length consensus amino acid sequence as shown in SEQ ID NO: 14. Some embodiments relate to immunogenic proteins having an amino acid sequence that is 94% identical to the full-length consensus amino acid sequence as shown in SEQ ID NO: 14. Some embodiments relate to immunogenic proteins having an amino acid sequence that is 95% identical to the full-length consensus amino acid sequence as shown in SEQ ID NO: 14. Some embodiments relate to immunogenic proteins having an amino acid sequence that is 96% identical to the full-length consensus amino acid sequence as shown in SEQ ID NO: 14. Some embodiments relate to immunogenic proteins having an amino acid sequence that is 97% identical to the full-length consensus amino acid sequence as shown in SEQ ID NO: 14. Some embodiments relate to immunogenic proteins having an amino acid sequence that is 98% identical to the full-length consensus amino acid sequence as shown in SEQ ID NO: 14. Some embodiments relate to immunogenic proteins having an amino acid sequence that is 99% identical to the full-length consensus amino acid sequence as shown in SEQ ID NO: 14.

[0323] In some embodiments, the protein does not contain a leader sequence. In some embodiments, the protein does not contain an IgE leader sequence. A fragment of the consensus protein can comprise at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% of the consensus protein. An immunogenic fragment of SEQ ID NO: 14 can be provided. The immunogenic fragment may comprise at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% of SEQ ID NO: 14. In some embodiments, the fragment includes a leader sequence, such as, for example, an immunoglobulin leader sequence, such as an IgE leader sequence. In some embodiments, the fragment does not contain a leader sequence. In some embodiments, the fragment does not contain a leader sequence, such as, for example, an IgE leader sequence.

[0324] Immunogenic fragments of proteins having amino acid sequences homologous to immunogenic fragments of SEQ ID NO: 14 can be provided. Such immunogenic fragments can comprise at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% of a protein having 95% or greater homology to SEQ ID NO: 14. Some embodiments relate to immunogenic fragments having 96% homology to immunogenic fragments of the consensus protein sequences herein. Some embodiments relate to immunogenic fragments having 97% homology to immunogenic fragments of the consensus protein sequences herein. Some embodiments relate to immunogenic fragments having 98% homology to an immunogenic fragment of a consensus protein sequence herein. Some embodiments relate to immunogenic fragments having 99% homology to an immunogenic fragment of a consensus protein sequence herein. In some embodiments, the fragment includes a leader sequence, such as, for example, an immunoglobulin leader sequence, such as an IgE leader sequence. In some embodiments, the fragment does not contain a leader sequence. In some embodiments, the fragment does not contain a leader sequence, such as, for example, an IgE leader sequence.

[0325] Immunogenic fragments of proteins having an amino acid sequence identical to an immunogenic fragment of SEQ ID NO: 14 can be provided. Such immunogenic fragments can comprise at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% of a protein having 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to the amino acid sequence shown in SEQ ID NO: 14. In some embodiments, the fragments include a leader sequence, such as, for example, an immunoglobulin leader sequence, such as an IgE leader sequence. In some embodiments, the fragments do not contain a leader sequence. In some embodiments, the fragment does not contain a leader sequence, such as, for example, an IgE leader sequence.

[0326] As indicated herein with respect to linking a signal peptide or leader sequence to the N-terminus of a protein, the signal peptide / leader sequence replaces the protein's N-terminal methionine that is encoded by the start codon of the nucleic acid sequence encoding the protein without the signal peptide coding sequence.

[0327] (8)NY-ESO-2

[0328] The immunogenic compositions of the present invention may comprise the cancer antigen New York Esop...

Claims

1. An immunogenic composition comprising a nucleic acid molecule encoding a consensus mouse TERT antigen, wherein the consensus mouse TERT antigen comprises an amino acid sequence selected from the group consisting of: a) an amino acid sequence selected from the group consisting of SEQ ID NO: 50 and SEQ ID NO: 52, b) an amino acid sequence having 95% or greater identity to an amino acid sequence selected from the group consisting of SEQ ID NO:50 and SEQ ID NO:52, and c) a fragment of an amino acid sequence selected from the group consisting of SEQ ID NO: 50 and SEQ ID NO: 52, wherein the fragment comprises at least 95% of the full-length amino acid sequence.

2. The immunogenic composition of claim 1, wherein the immunogenic composition further comprises one or more nucleotide sequences encoding one or more amino acid sequences selected from the group consisting of: a) an amino acid sequence selected from the group consisting of SEQ ID NO:2, SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:8, SEQ ID NO:10, SEQ ID NO:12, SEQ ID NO:14, SEQ ID NO:16, SEQ ID NO:18, SEQ ID NO:20, SEQ ID NO:22, SEQ ID NO:24, SEQ ID NO:58, SEQ ID NO:60, SEQ ID NO:62, SEQ ID NO:64, SEQ ID NO:66, SEQ ID NO:68, SEQ ID NO:70; b) an amino acid sequence having 95% or greater identity to an amino acid sequence selected from the group consisting of SEQ ID NO:2, SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:8, SEQ ID NO:10, SEQ ID NO:12, SEQ ID NO:14, SEQ ID NO:16, SEQ ID NO:18, SEQ ID NO:20, SEQ ID NO:22, SEQ ID NO:24, SEQ ID NO:58, SEQ ID NO:60, SEQ ID NO:62, SEQ ID NO:64, SEQ ID NO:66, SEQ ID NO:68, SEQ ID NO:70, and ... c) a fragment of an amino acid sequence selected from the group consisting of SEQ ID NO:2, SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:8, SEQ ID NO:10, SEQ ID NO:12, SEQ ID NO:14, SEQ ID NO:16, SEQ ID NO:18, SEQ ID NO:20, SEQ ID NO:22, SEQ ID NO:24, SEQ ID NO:58, SEQ ID NO:60, SEQ ID NO:62, SEQ ID NO:64, SEQ ID NO:66, SEQ ID NO:68, SEQ ID NO:70, wherein the fragment comprises at least 95% of the full-length amino acid sequence.

3. The immunogenic composition of claim 1, further comprising a nucleic acid encoding one or more antigens selected from the group consisting of MAGE A1, gp100, viral antigens, and combinations thereof.

4. The immunogenic composition of claim 3, wherein the viral antigen is an antigen from hepatitis B virus (HBV), hepatitis C virus (HCV) or human papillomavirus (HPV).

5. The immunogenic composition of claim 4, wherein the HBV antigen is HBV core antigen or HBV surface antigen or a combination thereof.

6. The immunogenic composition of claim 5, wherein the HCV antigen is HCV NS34A antigen, HCV NS5A antigen, HCV NS5B antigen, HCV NS4B antigen, or a combination thereof.

7. The immunogenic composition of claim 4, wherein the HPV antigen is HPV type 6 E6 antigen, HPV type 6 E7 antigen, HPV type 11 E6 antigen, HPV type 11 E7 antigen, HPV type 16 E6 antigen, HPV type 16 E7 antigen, HPV type 18 E6 antigen, HPV type 18 E7 antigen, or a combination thereof.

8. The immunogenic composition of claim 1, further comprising an immune checkpoint inhibitor.

9. The immunogenic composition of claim 1, wherein the nucleic acid molecule comprises a nucleotide sequence selected from the group consisting of: a) a nucleotide sequence selected from the group consisting of SEQ ID NO:49 and SEQ ID NO:51, b) a nucleotide sequence having 95% or greater identity with a nucleotide sequence selected from the group consisting of SEQ ID NO:49 and SEQ ID NO:51, and c) a fragment of a nucleotide sequence selected from the group consisting of SEQ ID NO: 49 and SEQ ID NO: 51, wherein the fragment comprises at least 95% of the full-length nucleotide sequence.

10. The immunogenic composition of claim 2, wherein the immunogenic composition comprises one or more nucleotide sequences selected from the group consisting of: a) a nucleotide sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 3, SEQ ID NO: 5, SEQ ID NO: 7, SEQ ID NO: 9, SEQ ID NO: 11, SEQ ID NO: 13, SEQ ID NO: 15, SEQ ID NO: 17, SEQ ID NO: 19, SEQ ID NO: 21, SEQ ID NO: 23, SEQ ID NO: 57, SEQ ID NO: 59, SEQ ID NO: 61, SEQ ID NO: 63, SEQ ID NO: 65, SEQ ID NO: 67 and SEQ ID NO: 69; b) a nucleotide sequence having 95% or greater identity to a nucleotide sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 3, SEQ ID NO: 5, SEQ ID NO: 7, SEQ ID NO: 9, SEQ ID NO: 11, SEQ ID NO: 13, SEQ ID NO: 15, SEQ ID NO: 17, SEQ ID NO: 19, SEQ ID NO: 21, SEQ ID NO: 23, SEQ ID NO: 57, SEQ ID NO: 59, SEQ ID NO: 61, SEQ ID NO: 63, SEQ ID NO: 65, SEQ ID NO: 67, and SEQ ID NO: 69, and c) a fragment of a nucleotide sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:5, SEQ ID NO:7, SEQ ID NO:9, SEQ ID NO:11, SEQ ID NO:13, SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO:19, SEQ ID NO:21, SEQ ID NO:23, SEQ ID NO:57, SEQ ID NO:59, SEQ ID NO:61, SEQ ID NO:63, SEQ ID NO:65, SEQ ID NO:67 and SEQ ID NO:69, wherein the fragment comprises at least 95% of the full-length nucleotide sequence.

11. The immunogenic composition of claim 1, wherein the nucleic acid is a plasmid.

12. The immunogenic composition of claim 1, wherein the composition comprises one or more plasmids.

13. The immunogenic composition of claim 1, further comprising an adjuvant.

14. The immunogenic composition of claim 13, wherein the adjuvant is IL-12, IL-15, IL-28 or RANTES.

15. A method of treating or preventing cancer in a subject in need thereof, the method comprising administering to the subject the immunogenic composition of claim 1.

16. The method of claim 15, wherein administering comprises an electroporation step.

17. The method of claim 15, further comprising administering an immune checkpoint inhibitor to the subject.

18. The method of claim 17, wherein the immunogenic composition and the immune checkpoint inhibitor are administered to the subject in a single formulation.

19. The method of claim 17, wherein the immunogenic composition and the immune checkpoint inhibitor are administered separately to the subject.

20. The method of claim 15, wherein the cancer is selected from the group consisting of melanoma, head and neck cancer, prostate cancer, liver cancer, cervical cancer, recurrent respiratory papillomatosis (RRP), anal cancer, blood cancer, ovarian cancer, and combinations thereof.

21. A nucleic acid molecule encoding a mouse TERT antigen, comprising one or more nucleotide sequences selected from the group consisting of: a) a nucleotide sequence selected from the group consisting of SEQ ID NO:49 and SEQ ID NO:51, b) a nucleotide sequence having 95% or greater identity with a nucleotide sequence selected from the group consisting of SEQ ID NO:49 and SEQ ID NO:51, and c) a fragment of a nucleotide sequence selected from the group consisting of SEQ ID NO: 49 and SEQ ID NO: 51, wherein the fragment comprises at least 95% of the full-length nucleotide sequence.

22. The nucleic acid molecule according to claim 21, further comprising one or more nucleotide sequences selected from the group consisting of: a) a nucleotide sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 3, SEQ ID NO: 5, SEQ ID NO: 7, SEQ ID NO: 9, SEQ ID NO: 11, SEQ ID NO: 13, SEQ ID NO: 15, SEQ ID NO: 17, SEQ ID NO: 19, SEQ ID NO: 21, SEQ ID NO: 23, SEQ ID NO: 57, SEQ ID NO: 59, SEQ ID NO: 61, SEQ ID NO: 63, SEQ ID NO: 65, SEQ ID NO: 67 and SEQ ID NO: 69; b) a nucleotide sequence having 95% or greater identity to a nucleotide sequence selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 3, SEQ ID NO: 5, SEQ ID NO: 7, SEQ ID NO: 9, SEQ ID NO: 11, SEQ ID NO: 13, SEQ ID NO: 15, SEQ ID NO: 17, SEQ ID NO: 19, SEQ ID NO: 21, SEQ ID NO: 23, SEQ ID NO: 57, SEQ ID NO: 59, SEQ ID NO: 61, SEQ ID NO: 63, SEQ ID NO: 65, SEQ ID NO: 67, and SEQ ID NO: 69, and c) a fragment of a nucleotide sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:5, SEQ ID NO:7, SEQ ID NO:9, SEQ ID NO:11, SEQ ID NO:13, SEQ ID NO:15, SEQ ID NO:17, SEQ ID NO:19, SEQ ID NO:21, SEQ ID NO:23, SEQ ID NO:57, SEQ ID NO:59, SEQ ID NO:61, SEQ ID NO:63, SEQ ID NO:65, SEQ ID NO:67 and SEQ ID NO:69, wherein the fragment comprises at least 95% of the full-length nucleotide sequence.

23. The nucleic acid molecule of claim 21, wherein the nucleic acid molecule is a plasmid.

24. An amino acid molecule of mouse TERT antigen, comprising one or more amino acid sequences selected from the group consisting of: a) an amino acid sequence selected from the group consisting of SEQ ID NO: 50 and SEQ ID NO: 52, b) an amino acid sequence having 95% or greater identity to an amino acid sequence selected from the group consisting of SEQ ID NO:50 and SEQ ID NO:52, and c) a fragment of an amino acid sequence selected from the group consisting of SEQ ID NO: 50 and SEQ ID NO: 52, wherein the fragment comprises at least 95% of the full-length amino acid sequence.

25. The amino acid molecule of claim 24, further comprising one or more amino acid sequences selected from the group consisting of: a) an amino acid sequence selected from the group consisting of SEQ ID NO:2, SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:8, SEQ ID NO:10, SEQ ID NO:12, SEQ ID NO:14, SEQ ID NO:16, SEQ ID NO:18, SEQ ID NO:20, SEQ ID NO:22, SEQ ID NO:24, SEQ ID NO:58, SEQ ID NO:60, SEQ ID NO:62, SEQ ID NO:64, SEQ ID NO:66, SEQ ID NO:68, SEQ ID NO:70; b) an amino acid sequence having 95% or greater identity to an amino acid selected from the group consisting of SEQ ID NO:2, SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:8, SEQ ID NO:10, SEQ ID NO:12, SEQ ID NO:14, SEQ ID NO:16, SEQ ID NO:18, SEQ ID NO:20, SEQ ID NO:22, SEQ ID NO:24, SEQ ID NO:58, SEQ ID NO:60, SEQ ID NO:62, SEQ ID NO:64, SEQ ID NO:66, SEQ ID NO:68, SEQ ID NO:70, and ... c) a fragment of an amino acid sequence selected from the group consisting of SEQ ID NO:2, SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:8, SEQ ID NO:10, SEQ ID NO:12, SEQ ID NO:14, SEQ ID NO:16, SEQ ID NO:18, SEQ ID NO:20, SEQ ID NO:22, SEQ ID NO:24, SEQ ID NO:58, SEQ ID NO:60, SEQ ID NO:62, SEQ ID NO:64, SEQ ID NO:66, SEQ ID NO:68, SEQ ID NO:70, wherein the fragment comprises at least 95% of the full-length amino acid sequence.

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