TERT immunogenic compositions and methods of treatment using the same

Abstract

To provide methods for treating cancer and in particular immunogenic compositions that treat and provide protection against tumor.SOLUTION: The present invention discloses compositions comprising optimized consensus TERT antigens.SELECTED DRAWING: None

Description

Technical Field

[0001] (Cross - Reference to Related Applications) This application claims priority to U.S. Provisional Patent Application No. 62 / 402,695, filed on September 30, 2016, and U.S. Provisional Patent Application No. 62 / 468,124, filed on March 7, 2017, the entire contents of which are incorporated herein by reference.

[0002] Disclosed herein are compositions and methods for treating cancer, and in particular, immunogenic compositions that treat tumor growth and provide protection against tumor growth.

Background Art

[0003] Cancer is one of the leading causes of death worldwide and in the United States, ranking second among the most common causes of death and accounting for approximately one - quarter of all deaths. Cancer originates from a single cell that has changed from a normal cell to a tumor cell. Such changes are often a multi - step process that progresses from a pre - cancerous lesion to a malignant tumor. This progression involves many factors, including aging, genetic contributions, and exposure to external causative agents such as physical carcinogens (e.g., ultraviolet and ionizing radiation), chemical carcinogens (e.g., asbestos, components of tobacco smoke), and biological carcinogens (e.g., certain viruses, bacteria, and parasites).

[0004] The prevention, diagnosis, and treatment of cancer can take many different forms. Prevention can include screening for predisposing factors (e.g., specific gene variants), modifying habits (e.g., smoking, diet, and exercise levels), and vaccination against viruses (e.g., human papillomavirus, hepatitis B virus). Treatments can include chemotherapy, radiation therapy, and surgical removal of tumors or cancerous tissue. Despite the availability of numerous prevention and treatment methods, such methods often achieve only limited success in the effective prevention and / or treatment of common cancers.

[0005] Accordingly, there is a need for the identification and development of cancer prevention and / or treatment compositions and cancer prevention and / or treatment methods to facilitate the clinical management of protection against disease progression. Further, in subjects suffering from cancer, more effective treatments are sought to delay disease progression and / or reduce mortality.

SUMMARY OF THE INVENTION

[0006] 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 at least 95% identity with 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, the fragment comprising at least 95% of the full-length amino acid sequence.

[0007] 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 at least 95% identity with 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, the fragment comprising at least 95% of the full-length amino acid sequence.

[0008] 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 derived from hepatitis B virus (HBV), hepatitis C virus (HCV), or human papillomavirus (HPV). In one embodiment, the HBV antigen is the HBV core antigen or the HBV surface antigen, or a combination thereof. In one embodiment, the HCV antigen is the HCV NS3 / 4A antigen, the HCV NS5A antigen, the HCV NS5B antigen, the HCV NS4B antigen, or a combination thereof. In one embodiment, the HPV antigen is the HPV type 6 E6 antigen, the HPV type 6 E7 antigen, the HPV type 11 E6 antigen, the HPV type 11 E7 antigen, the HPV type 16 E6 antigen, the HPV type 16 E7 antigen, the HPV type 18 E6 antigen, the HPV type 18 E7 antigen, or a combination thereof.

[0009] In one embodiment, the immunogenic composition comprises an immune checkpoint inhibitor.

[0010] 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 at least 95% identity with 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.

[0011] In one embodiment, the immunogenic composition 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 at least 95% identity with 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, the fragment comprising at least 95% of the full-length nucleotide sequence.

[0012] In one embodiment, the nucleic acid molecule is a plasmid.

[0013] In one embodiment, the immunogenic composition comprises one or more plasmids.

[0014] In one embodiment, the immunogenic composition further comprises an adjuvant. In one embodiment, the adjuvant is IL-12, IL-15, IL-28, or RANTES.

[0015] In one embodiment, the present invention relates to a method for treating or preventing cancer in a subject in need of cancer treatment or prevention, the method comprising administering to the subject an immunogenic composition comprising a nucleic acid molecule encoding a consensus TERT antigen. In one embodiment, the method of administration comprises an electroporation step. In one embodiment, the method further comprises administering an immune checkpoint inhibitor to the subject.

[0016] 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.

[0017] 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.

[0018] 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 at least 95% identity with 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, the fragment comprising at least 95% of the full-length nucleotide sequence.

[0019] 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 at least 95% identity with 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, the fragment comprising at least 95% of the full-length nucleotide sequence.

[0020] In one embodiment, the nucleic acid molecule is a plasmid.

[0021] 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 at least 95% identity with 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, the fragment comprising at least 95% of the full-length amino acid sequence.

[0022] 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 at least 95% identity with 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, the fragment comprising at least 95% of the full-length amino acid sequence. BRIEF DESCRIPTION OF THE DRAWINGS

[0023]

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Mode for Carrying Out the Invention

[0024] The present invention relates to an immunogenic composition comprising a synthetic consensus TERT antigen, alone or in combination with at least one additional cancer antigen or an inhibitor of an immune checkpoint protein, and a method of using the composition for treating a disease or disorder. In one embodiment, the immunogenic composition comprises TERT and at least one additional cancer antigen consensus sequence. Cancer antigen consensus sequences that can be included in the immunogenic composition include, but are not limited to, tyrosinase (Tyr), melanoma preferentially expressed antigen (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 of the prostate (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 of cancer prevention or treatment.

[0025] One method of designing the above nucleic acid and the amino acid sequence of the corresponding recombinant cancer antigen encoded thereby is to introduce mutations that alter specific amino acids in the overall amino acid sequence of the native cancer antigen. The introduction of mutations does not greatly modify the cancer antigen to the extent that the cancer antigen cannot be universally applied among mammalian subjects. For example, it can be applied to human or canine subjects. However, the introduction of such mutations changes the antigen to an extent that the resulting amino acid sequence exceeds the tolerance range, i.e., is changed sufficiently to be regarded as a foreign antigen for eliciting an immune response. Another method could be to create a recombinant cancer antigen having at least 85% to 99% amino acid sequence identity, at least 90% to 98% sequence identity, at least 93% to 98% sequence identity, or at least 95% to 98% sequence identity to the corresponding native cancer antigen. In some examples, the above recombinant cancer antigen has 95%, 96%, 97%, 98% or 99% amino acid sequence identity to the corresponding native cancer antigen. The above native cancer antigen is an antigen normally involved in a specific cancer or tumor. Depending on the cancer antigen, the consensus sequence of the cancer antigen can span across mammals, or within a subclass of the class, or across an entire viral strain or serotype. Some cancer antigens do not vary significantly from the wild-type of the amino acid sequence of the cancer antigen. Some cancer antigens have significantly different nucleic acid / amino acid sequences among species and thus cannot give rise to a consensus sequence. In these examples, a recombinant cancer antigen that elicits an immune response beyond the tolerance range is generated when the recombinant cancer antigen has at least 85% to 99% amino acid sequence identity, at least 90% to 98% sequence identity, at least 93% to 98% sequence identity, at least 95% to 98% sequence identity to the corresponding native cancer antigen. In some examples, the above recombinant cancer antigen has 95%, 96%, 97%, 98% or 99% amino acid sequence identity to the corresponding native cancer antigen. The above-mentioned efforts can be combined such that the final recombinant cancer antigen has similarity expressed as a percentage to the amino acid sequence of the unmutated cancer antigen as discussed above.

[0026] The recombinant cancer antigen described above can induce a response of antigen-specific T cells and / or high-titer antibodies, thereby inducing or eliciting an immune response directed against 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-gamma (IFN-γ) and / or tumor necrosis factor alpha (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 a tumor or cancer expressing the antigen, such as, but not limited to, factors that downregulate presentation by MHC, antigen-specific regulatory T cells (Tregs), cytokines such as PD-L1, FasL, IL-10, and TGF-β, tumor-associated macrophages, tumor-associated fibroblasts, soluble factors produced by immunosuppressive cells, factors that upregulate CTLA-4, PD-1, MDSC, MCP-1, and immune checkpoint molecules.

[0027] The immunogenic composition may further be combined with antibodies against checkpoint inhibitors such as PD-1 and PDL-1 to increase the stimulation of both cellular and humoral immune responses. Using anti-PD-1 or anti-PDL-1 antibodies prevents PD-1 or PDL-1 from suppressing the response of T cells and / or B cells. Overall, designing cancer antigens recognized by the immune system helps overcome other forms of immunosuppression by cancer cells, and these immunogenic compositions can be used in combination with inhibitory or inhibitory therapies (such as anti-PD-1 and anti-PDL-1 antibody therapies) to further increase T cell and / or B cell responses.

[0028] 1. Definitions Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. In case of conflict, the present specification, including definitions, will control. Although methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention, exemplary methods and materials are described below. All publications, patent applications, patents, and other references cited herein are incorporated by reference in their entirety. It should be noted that the materials, methods, and examples disclosed herein are merely exemplary and are not intended to be limiting. The terms used herein are for the purpose of describing particular embodiments only and are not intended to be limiting.

[0029] As used herein, the terms "comprise(s)", "include(s)", "having", "has", "can", "contain(s)", and variations thereof are intended to be transitional phrases, terms, or words that do not preclude the possibility of additional acts or structures and are not limiting. The singular forms "a", "and", and "the" may also have a plural meaning unless the context clearly dictates otherwise. The present disclosure contemplates other embodiments that "comprise", "consist of", and "consist essentially of" the embodiments or elements shown herein, whether or not explicitly described.

[0030] Regarding the description of numerical ranges herein, each number falling therebetween with the same degree of precision is explicitly contemplated. For example, in the case of the range 6 - 9, the numbers 7 and 8 in addition to 6 and 9 are contemplated, and in the case of 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 explicitly contemplated.

[0031] As used herein, "adjuvant" means any molecule added to an immunogenic composition described herein to enhance the immunogenicity of a nucleic acid molecule and an antigen encoded by a coding nucleic acid sequence described below in this specification.

[0032] As used herein, "antibody" means an antibody of class IgG, IgM, IgA, IgD, or IgE, or a fragment or derivative thereof (including Fab, F(ab’)2, Fd), as well as single-chain antibodies, diabodies, bispecific antibodies, bifunctional antibodies, and derivatives thereof. The antibody can be an antibody isolated from a mammalian serum sample, a polyclonal antibody, an affinity-purified antibody, or a mixture thereof, provided that it exhibits sufficient binding specificity for a desired epitope or a sequence derived therefrom.

[0033] As used herein, "coding sequence" or "coding nucleic acid" means a nucleic acid (RNA or DNA molecule) containing a nucleotide sequence encoding a protein. The coding sequence can further include start and stop signals, and is operably linked to regulatory elements including a promoter and a polyadenylation signal capable of inducing expression in a cell of an individual or mammal to which the nucleic acid is administered.

[0034] As used herein, "complement" or "complementary" means that a nucleic acid can form Watson-Crick (e.g., A- / U and C-G) or Hoogsteen-type base pairs between nucleotides or nucleotide analogs of a nucleic acid molecule.

[0035] As used herein, "consensus" or "consensus sequence" means a polypeptide sequence based on the alignment analysis of multiple sequences for the same gene from different organisms. A nucleic acid sequence encoding a consensus polypeptide sequence can be prepared. An immunogenic composition containing a protein comprising a consensus sequence and / or a nucleic acid molecule encoding such a protein can be used to induce broad immunity against an antigen. As used interchangeably herein, "electroporation," "electropermeabilization," "electrokinetic enhancement" ("EP") mean the use of transmembrane electric field pulses to create micropores (pores) in biological membranes. The presence of such micropores enables biomolecules such as plasmids, oligonucleotides, siRNA, drugs, ions, and water to pass from one side of the cell membrane to the other side.

[0036] As used herein, "fragment" with respect to a nucleic acid sequence means a nucleic acid sequence that encodes a polypeptide capable of inducing an immune response in a mammal that cross-reacts with the antigens described herein, or a part thereof. The above fragment can be a DNA fragment selected from at least one of various nucleotide sequences encoding the protein fragments described below. The 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 of the nucleic acid sequences described below. In some embodiments, the fragment comprises 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, at least 1000 nucleotides or more of at least one of the nucleic acid sequences described below.

[0037] "Fragment" or "immunogenic fragment" as used with respect to a polypeptide sequence means a polypeptide capable of inducing an immune response in a mammal that cross-reacts with the antigens described herein. The fragment can be a polypeptide fragment selected from at least one of the following various amino acid sequences. 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 is 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 of the protein sequences disclosed herein.

[0038] As used herein, the term "gene construct" refers to a DNA or RNA molecule containing a nucleotide sequence encoding a protein. The coding sequence includes initiation and termination signals and is operably linked to regulatory elements including a promoter and a polyadenylation signal capable of inducing expression in the cells of an individual to which the nucleic acid molecule is administered. As used herein, the term "expressible form" refers to a gene construct containing the regulatory elements necessary to be operably linked to a coding sequence encoding a protein such that the coding sequence is expressed when present in the cells of an individual.

[0039] 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 the hybridization of a completely complementary sequence to a target nucleic acid is represented by the functional term "substantially homologous". When used with respect to a double-stranded nucleic acid sequence such as a cDNA or genomic clone, the term "substantially homologous" as used herein refers to a probe that can hybridize to the strand of the double-stranded nucleic acid sequence under low stringency conditions. When used with respect to a single-stranded nucleic acid sequence, the term "substantially homologous" as used herein refers to a probe that can hybridize to a single-stranded nucleic acid template sequence under low stringency conditions (i.e., is complementary to the sequence).

[0040] As used herein, "identical" or "identity" in the context of two or more nucleic acid or polypeptide sequences means that the sequences have a specified percentage of identical residues over a specified region. To calculate this percentage, the two sequences are optimally aligned, the two sequences are compared over the specified region, the number of positions at which the identical residues of the two sequences occur 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 percent value of sequence identity. If the lengths of the two sequences are different, or if one or more attached ends are generated by the alignment and only a single sequence is included in the specified comparison region, the residues of the single sequence are included in the denominator but not in the numerator of the calculation. When comparing DNA and RNA, thymine (T) and uracil (U) can be considered equivalent. The calculation of identity can be performed manually or using computer sequence algorithms such as BLAST, BLAST 2.0, etc.

[0041] As used herein, "immune response" means that the host immune system, such as the mammalian immune system, is activated in response to the introduction of an antigen. This immune response can be in cellular or humoral form, or both.

[0042] As used herein, "nucleic acid" or "oligonucleotide" or "polynucleotide" means at least two nucleotides covalently linked to each other. By representing a single strand, the sequence of the complementary strand is also defined. Thus, a nucleic acid includes the complementary strand of the single strand being represented. Many variants of a given nucleic acid can be used for the same purpose as the given nucleic acid. Thus, a nucleic acid includes substantially identical nucleic acids and their complements. A single strand provides a probe that can hybridize to a target sequence under stringent hybridization conditions. Thus, a nucleic acid includes probes that hybridize under stringent hybridization conditions.

[0043] The nucleic acid can be single-stranded or double-stranded, and can also contain a part of the sequences of both double-stranded and single-stranded. The nucleic acid can be DNA, genomic DNA, cDNA, RNA, or a hybrid, and the nucleic acid can contain a combination of deoxyribonucleotides and ribonucleotides, and can contain a combination of bases including uracil, adenine, thymine, cytosine, guanine, inosine, xanthine, hypoxanthine, isocytosine, and isoguanine. The nucleic acid can be obtained by chemical synthesis methods or recombinant methods.

[0044] As used herein, "functionally linked" means that a gene is expressed under the control of a promoter that is spatially connected to the gene. The 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 gene controlled by the promoter and the promoter in the gene from which the promoter is derived. As is known in the art, changes in this distance can be accommodated without loss of promoter function.

[0045] As used herein, "peptide", "protein", or "polypeptide" can mean a linked sequence of amino acids, and can be natural, synthetic, or a modification of natural and synthetic or a combination thereof.

[0046] As used herein, a "promoter" refers to a synthetic or naturally occurring molecule capable of conferring, activating, or enhancing expression of a nucleic acid in a cell. A promoter can contain one or more specific transcriptional regulatory sequences to further enhance expression and / or modify its spatial and / or temporal expression. A promoter can also contain distal enhancer or repression elements, which can be located as far as thousands of base pairs from the transcription start site. Promoters can be derived from sources including viruses, bacteria, fungi, plants, insects, and animals. A promoter can regulate the expression of genetic components constitutively or differentially with respect to the cell, tissue, or organ in which expression occurs, or with respect to the developmental stage in which expression occurs, or can regulate the expression of genetic components in response to external stimuli, such as physiological stress, pathogens, metal ions, inducers, etc. Representative examples of promoters include a bacteriophage T7 promoter, a bacteriophage T3 promoter, an SP6 promoter, a lac operator promoter, a tac promoter, an SV40 late promoter, an SV40 early promoter, an RSV-TR promoter, a CMV IE promoter, an SV40 early promoter, an SV40 late promoter, and a CMV IE promoter.

[0047] "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. Generally, a signal peptide / leader sequence directs the localization of a protein. As used herein, a signal peptide / leader sequence can facilitate secretion of a protein from the cell in which it is produced. The signal peptide / leader sequence is often cleaved from the rest of the protein (often referred to as the mature protein) upon secretion from the cell. The signal peptide / leader sequence is attached to the amino terminus (i.e., N-terminus) of a protein.

[0048] As used herein, "stringent hybridization conditions" refers to conditions under which, for example, in a complex mixture of nucleic acids, a first nucleic acid sequence (e.g., a probe) hybridizes to a second nucleic acid sequence (e.g., a target). Since stringent conditions are sequence-dependent, they vary depending on the circumstances. Stringent conditions can be selected to be about 5-10 °C lower than the melting temperature (Tm) for a particular sequence at a given ionic strength and pH. Tm can be defined as the temperature at which 50% of the probe complementary to the target (under defined ionic strength, pH, and nucleic acid concentration) hybridizes to the target sequence at equilibrium (since the target sequence is present in excess, at Tm, 50% of the probe is occupied at equilibrium). Stringent conditions can have a sodium ion concentration of less than about 1.0 M at pH 7.0-8.3, e.g., a sodium ion concentration of about 0.01-1.0 M (or other salts), and a temperature of at least about 30 °C for short probes (e.g., 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 the addition of destabilizing agents such as formamide. For selective or specific hybridization, the positive signal can be at least 2-10 times that of background hybridization. Examples of stringent hybridization conditions include the following: incubate at 42 °C in 50% formamide, 5× SSC, and 1% SDS. Or, incubate at 65 °C in 5× SSC, 1% SDS, and wash at 65 °C in 0.2× SSC, 0.1% SDS.

[0049] As used herein, "subject" means a mammal that desires or requires immunization with the immunogenic composition described herein. The mammal can be a human, chimpanzee, dog, cat, horse, cow, mouse, or rat.

[0050] As used herein, "substantially complementary" means that over a region of 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, 450, 540 or more nucleotides or amino acids, the first sequence is 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 to the complement of the second sequence, or that the two sequences hybridize under stringent hybridization conditions.

[0051] As used herein, "substantially identical" means that over a region of 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, 450, 540 or more nucleotides or amino acids, the first sequence and the second sequence 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, or, with respect to nucleic acids, when the first sequence is substantially complementary to the complement of the second sequence.

[0052] As used herein, "treatment" or "treating" can mean protecting an animal from a disease by preventing, suppressing, repressing, or completely eliminating the disease. Preventing a disease involves administering an immunogenic composition of the invention to an animal before the onset of the disease. Suppressing a disease involves administering an immunogenic composition of the invention to an animal after induction of the disease but before the clinical appearance of the disease. Suppressing a disease involves administering an immunogenic composition of the invention to an animal after the clinical appearance of the disease.

[0053] As used herein, "variant" with respect to nucleic acids 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 that is 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 substantially identical sequence thereof.

[0054] As used with respect to a peptide or polypeptide, a "variant" has an amino acid sequence that differs by amino acid insertions, deletions, or conservative substitutions, but retains at least one biological activity. A variant can also mean a protein having an amino acid sequence that is substantially identical to a reference protein and that retains at least one biological activity. Conservative substitution of an amino acid, i.e., substitution of one amino acid for another with similar properties (e.g., hydrophilicity, degree, and distribution of charged regions), is generally recognized in the art as involving minor changes. Such minor changes can be partially identified by considering the hydropathic index of the amino acids, as understood in the art. Kyte et al., J. Mol. Biol. 157:105-132 (1982). The hydropathic index of an amino acid is based on consideration of its hydrophobicity and charge. Amino acids with similar hydropathic indices can be substituted, and it is known in the art that protein function can still be maintained thereafter. In one aspect, amino acids having a hydropathic index of ±2 are substituted. Substitutions that result in a protein retaining a biological function can also be identified using the hydrophilicity of the amino acids. By considering the hydrophilicity of amino acids in relation to a peptide, the local maximum average hydrophilicity of the peptide can be calculated, which is a useful measure that has been reported to correlate well with antigenicity and immunogenicity. U.S. Patent No. 4,554,101 (this document is incorporated herein by reference in its entirety). As understood in the art, substituting amino acids with similar hydrophilicity values results in a peptide that retains biological activities such as immunogenicity. Substitutions can be made using amino acids whose hydrophilicity values are within ±2 of each other. Both the hydrophobic index and the hydrophilicity value of an amino acid are affected by the specific side chain of that amino acid. Consistent with these findings, amino acid substitutions that are compatible with biological function are understood to depend on the relative similarity of the amino acids, particularly the relative similarity of the side chains of the amino acids, which is revealed by hydrophobicity, hydrophilicity, charge, size, and other properties.

[0055] The variant may be a nucleic acid sequence that is substantially identical over the entire length of the complete gene sequence or a fragment thereof. The nucleic acid sequence may be 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 gene sequence or a fragment thereof. The variant may be an amino acid sequence that is substantially identical over the entire length of the amino acid sequence or a fragment thereof. The amino acid sequence may be 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 or a fragment thereof.

[0056] As used herein, "vector" means a nucleic acid sequence containing an origin of replication. The vector can be a viral vector, bacteriophage, bacterial artificial chromosome, or yeast artificial chromosome. The vector can be a DNA vector or an RNA vector. The vector is a self-replicating episomal vector and, in one embodiment, an expression plasmid. The vector can contain or incorporate one or more non-homologous nucleic acid sequences.

[0057] 2. Synthetic consensus TERT The present invention provides an optimized consensus sequence of the TERT antigen. In one embodiment, the antigen encoded by the optimized consensus sequence can induce an immune response in a mammal. In one embodiment, the antigen encoded by the optimized consensus sequence can contain an epitope (s) that is particularly effective as an immunogen capable of inducing an immune response to the antigen.

[0058] The optimized consensus array can be a consensus array derived from two or more native TERT antigens. The optimized consensus array can have modifications for improving the consensus array and / or expression. Modifications include codon optimization, RNA optimization, addition of a Kozak sequence to enhance translation initiation efficiency, and / or addition of an immunoglobulin leader sequence to enhance immunogenicity. The TERT antigen encoded by the optimized consensus array can include a signal peptide such as, but not limited to, an immunoglobulin signal peptide including immunoglobulin E (IgE) or immunoglobulin (IgG) signal peptide. In some embodiments, the antigen encoded by the optimized consensus array can have a hemagglutinin (HA) tag. The TERT antigen encoded by the optimized consensus array can be designed to induce a stronger cellular and / or humoral immune response than the corresponding native antigen. The TERT antigen encoded by the optimized consensus array can be designed to break tolerance and exhibit synergistic effects with anti-cancer immunotherapy.

[0059] In one embodiment, the optimized consensus TERT is designed to break tolerance to native human TERT. In one embodiment, the sequence encoding human optimized consensus TERT is as set forth 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 antigen encoded by human optimized consensus TERT has an amino acid sequence as set forth 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.

[0060] In one embodiment, the optimized consensus TERT is designed to break tolerance to native mouse TERT. In one embodiment, the sequence encoding the mouse optimized consensus TERT is as set forth in SEQ ID NO: 49 or SEQ ID NO: 51. In one embodiment, the antigen encoded by the mouse optimized consensus TERT has the amino acid sequence as set forth in SEQ ID NO: 50 or SEQ ID NO: 52.

[0061] In one embodiment, the optimized consensus TERT is designed to break tolerance to native rhesus TERT. In one embodiment, the sequence encoding the rhesus optimized consensus TERT is as set forth in SEQ ID NO: 53 or SEQ ID NO: 55. In one embodiment, the antigen encoded by the mouse optimized consensus TERT has the amino acid sequence as set forth in SEQ ID NO: 54 or SEQ ID NO: 56.

[0062] In one embodiment, the TERT antigen encoded by the optimized consensus 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 sequences operably linked to the sequence encoding the leader of IgE are as set forth in SEQ ID NO: 45, SEQ ID NO: 49, and SEQ ID NO: 53. In one embodiment, the TERT antigen encoded by the optimized consensus operably linked to the leader sequence of IgE is as set forth in SEQ ID NO: 46, SEQ ID NO: 50, and SEQ ID NO: 54.

[0063] In one embodiment, the regulatory element is a start codon. Thus, in one embodiment, the invention relates to a nucleic acid sequence set forth 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 having a start codon at its 5' end. In one embodiment, the invention relates to an amino acid sequence set forth 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 (e.g., methionine) encoded by the N-terminal start codon.

[0064] In one embodiment, the regulatory element is at least one stop codon. Thus, in one embodiment, the present invention relates to a nucleic acid sequence set forth 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 homolog thereof, operably linked to a nucleotide sequence having 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.

[0065] In one embodiment, the optimized consensus sequence encoding the TERT antigen may encode a peptide having the amino acid sequence set forth 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 set forth 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 set forth 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 having at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity over the entire length of the amino acid sequence set forth 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.

[0066] In some embodiments, the optimized consensus TERT antigen may be encoded by RNA that is a transcription product from a DNA sequence having at least about 96%, 97%, 98%, 99%, or 100% identity over the entire length of the nucleic acid sequence set forth 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 may be encoded by RNA that encodes an amino acid sequence having at least about 96%, 97%, 98%, 99% or 100% identity over the entire length of the amino acid sequence set forth 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.

[0067] The optimized consensus code TERT antigen can be a peptide having the amino acid sequence set forth in SEQ ID NO: 2, SEQ ID NO: 4, SEQ ID NO: 6, or SEQ ID NO: 8. In some embodiments, it can have an amino acid sequence having at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity over the full length of the amino acid sequence set forth 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.

[0068] An immunogenic fragment of a protein having an amino acid sequence homologous to the 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 can be provided. Such an immunogenic fragment 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 that is 95% 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 an immunogenic fragment having 96% homology to an immunogenic fragment of the consensus protein sequence herein. Some embodiments relate to an immunogenic fragment having 97% homology to an immunogenic fragment of the consensus protein sequence herein. Some embodiments relate to an immunogenic fragment having 98% homology to an immunogenic fragment of the consensus protein sequence herein. Some embodiments relate to an immunogenic fragment having 99% homology to an immunogenic fragment of the consensus protein sequence herein. In some embodiments, the immunogenic fragment comprises a leader sequence, such as an immunoglobulin leader, such as an IgE leader. In some embodiments, the immunogenic fragment does not comprise a leader sequence.

[0069] In one embodiment, an immunogenic fragment of an optimized consensus TERT antigen encodes an optimized consensus TERT antigen that encodes at least one immunodominant or sub-immunodominant epitope of the full-length optimized consensus TERT antigen.

[0070] Some embodiments relate to an immunogenic 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 that comprises 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. 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 comprises a sequence encoding a leader sequence, such as an immunoglobulin leader, such as an IgE leader. In some embodiments, the fragment does not comprise a coding sequence encoding a leader sequence.

[0071] 3. Immunogenic composition In one embodiment, the immunogenic composition of the present invention can 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 the TTAGGG tag at the ends of telomeres to prevent cell death due to chromosome shortening. Hyperproliferative cells may exhibit abnormally high expression of hTERT. Aberrant expression of hTERT may also occur in hyperproliferative cells infected with HCV and HPV. Therefore, immunotherapy against HPV and HCV can be improved by targeting cells that express hTERT at abnormal levels. The HPV and HCV antigens are described in more detail below. In one aspect, the hTERT cancer antigen can be further defined by U.S. Patent Application No. 14 / 139,660, filed December 23, 2013, the entire contents of which are incorporated herein by reference.

[0072] 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 monkey TERT (rhTERT).

[0073] By expressing TERT in dendritic cells transfected with the TERT gene, antigen-specific CD8 + cytotoxic T cells can be induced, and CD4 + T cells can be elicited. Therefore, the use of hTERT expression in antigen-presenting cells (APCs) to delay aging and sustain the ability of APCs to present selected antigens can be used in immunotherapy, such as the methods described herein.

[0074] The TERT antigen can be associated with or expressed by any number 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 cancer. Thus, when the TERT antigen described herein is included, such a vaccine can be used to treat subjects suffering from any number 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 cancer.

[0075] The TERT antigen can induce or elicit an immune response that targets or is reactive against cancers or tumors that express the antigen by inducing antigen-specific T cells and / or a high-titer antibody response. In some embodiments, the induced or elicited immune response can be a cellular, humoral, or both cellular and humoral immune response. In some embodiments, the induced or elicited 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 that express antigens, including but not limited to factors that downregulate MHC presentation, factors that upregulate antigen-specific regulatory T cells (Tregs), cytokines such as PD-L1, FasL, IL-10, and TGF-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.

[0076] The TERT antigen can include protein epitopes that make the TERT antigen particularly effective as an immunogen that can induce an anti-TERT immune response against it. The TERT antigen can include the full-length translation product, variants thereof, fragments thereof, or combinations thereof. The TERT antigen can include a consensus protein.

[0077] The nucleic acid sequence encoding the TERT antigen or the consensus TERT antigen can be optimized with respect to codon usage and the corresponding RNA transcript. The nucleic acid encoding the TERT antigen or the consensus TERT antigen can be codon and RNA optimized for expression. In some embodiments, the nucleic acid sequence encoding the TERT antigen or the consensus TERT antigen can include a Kozak sequence (e.g., GCC ACC) to enhance translation efficiency. The nucleic acid encoding the TERT antigen or the consensus TERT antigen can include multiple stop codons (e.g., TGA TGA) to enhance the efficiency of translation termination.

[0078] The nucleic acid encoding the TERT antigen or the consensus TERT antigen may encode the leader sequence of immunoglobulin E (IgE). The nucleic acid encoding the TERT antigen or the consensus TERT antigen can further encode the 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 consensus TERT antigen by a peptide bond, respectively. The nucleic acid encoding the TERT antigen or the consensus TERT antigen may include the nucleotide sequence encoding the IgE leader sequence. In some embodiments, the nucleic acid encoding the TERT antigen or the consensus TERT antigen does not include or has no nucleotides encoding the IgE leader sequence.

[0079] In some embodiments, the nucleic acid encoding the TERT antigen or consensus TERT antigen may be a heterologous nucleic acid sequence and / or may have one or more heterologous nucleic acid sequences. The nucleic acid encoding the TERT antigen or consensus TERT antigen can be mutated relative to the wild-type TERT antigen such that one or more amino acids or residues in the amino acid sequence of the TERT antigen or consensus TERT antigen are each replaced with another amino acid or residue, i.e., substituted. By mutating the nucleic acid encoding the TERT antigen or consensus TERT antigen relative to the wild-type TERT antigen such that one or more residues in the amino acid sequence of the TERT antigen or consensus TERT antigen are each replaced with another residue, i.e., substituted, the immune system in the mammal to which the nucleic acid encoding the TERT antigen or consensus TERT antigen, the TERT antigen or consensus TERT antigen, or a combination thereof is administered can be made non-tolerant to TERT. The nucleic acid encoding the TERT antigen or consensus TERT antigen can be mutated relative to the wild-type TERT antigen such that both arginine 589, aspartic acid 1005, or both arginine 589 and aspartic acid 1005 in the amino acid sequence of the TERT antigen or consensus TERT antigen are replaced with tyrosine residues, i.e., substituted.

[0080] In one aspect, the TERT antigen may be the nucleic acid sequence of SEQ ID NO: 23 or SEQ ID NO: 57, which encode the amino acid sequences of 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 the IgE leader sequence. In one embodiment, the TERT antigen can be linked to the IgE leader sequence and an HA tag. In other embodiments, the TERT antigen may not include and / or not be linked to the IgE leader sequence and / or HA tag.

[0081] In some embodiments, the TERT antigen may be a nucleic acid sequence having 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% identity over the full length of the nucleic acid sequence set forth in SEQ ID NO: 23 or SEQ ID NO: 57. In other embodiments, the TERT antigen may be a nucleic acid sequence encoding an amino acid sequence having 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% identity over the full length of the amino acid sequence set forth in SEQ ID NO: 24 or SEQ ID NO: 58. The TERT antigen may be an amino acid sequence having 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% identity over the full length of the amino acid sequence set forth in SEQ ID NO: 24 or SEQ ID NO: 58.

[0082] 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 acids can be provided that encode immunogenic proteins having 95% or less identity, 96% or less identity, 97% or less identity, 98% or less identity, and 99% or less identity to the sequence. Similarly, nucleic acid sequences encoding immunogenic fragments described herein and immunogenic fragments of proteins homologous to the proteins described herein are provided.

[0083] Some embodiments relate to nucleic acid molecules encoding immunogenic proteins having 95% homology with the nucleic acid coding sequences herein. Some embodiments relate to nucleic acid molecules encoding immunogenic proteins having 96% homology with the nucleic acid coding sequences herein. Some embodiments relate to nucleic acid molecules encoding immunogenic proteins having 97% homology with the nucleic acid coding sequences herein. Some embodiments relate to nucleic acid molecules encoding immunogenic proteins having 98% homology with the nucleic acid coding sequences herein. Some embodiments relate to nucleic acid molecules encoding immunogenic proteins having 99% homology with the nucleic acid coding sequences herein. In some embodiments, a nucleic acid molecule having a coding sequence disclosed herein that is homologous to the coding sequence of the consensus protein disclosed herein comprises a sequence encoding an IgE leader sequence linked to the 5' end of the coding sequence encoding the homologous protein sequence disclosed herein.

[0084] Some embodiments relate to nucleic acid sequences encoding proteins having a specified identity (%) with the full-length TERT antigen, immunogenic fragments of the TERT antigen, and immunogenic fragments of proteins having identity with the TERT antigen. Such nucleic acid molecules can be provided that encode immunogenic proteins having 80% or less identity with the full-length TERT sequence, 85% or less identity with the full-length TERT sequence, 90% or less identity with the full-length TERT sequence, 91% or less identity with the full-length TERT sequence, 92% or less identity with the full-length TERT sequence, 93% or less identity with the full-length TERT sequence, 94% or less identity with the full-length TERT sequence, 95% or less identity with the full-length TERT sequence, 96% or less identity with the full-length TERT sequence, 97% or less identity with the full-length TERT sequence, 98% or less identity with the full-length TERT sequence, and 99% or less identity with the full-length TERT sequence. Similarly, nucleic acid sequences encoding immunogenic fragments described herein and immunogenic fragments of proteins having the same identity (%) as the TERT antigen described herein, as shown above, are also provided.

[0085] In some embodiments, the nucleic acid sequence does not include a coding sequence encoding a leader sequence. In some embodiments, the nucleic acid sequence does not include a coding sequence encoding an IgE leader.

[0086] Some embodiments relate to a fragment of SEQ ID NO: 23 or SEQ ID NO: 57. The fragment may 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 fragment may 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 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: 23 or SEQ ID NO: 57. In some embodiments, the fragment includes a sequence encoding a leader sequence, such as an immunoglobulin leader, such as an IgE leader. In some embodiments, the fragment does not include a coding sequence encoding a leader sequence. In some embodiments, the fragment does not include a coding sequence encoding a leader sequence such as an IgE leader.

[0087] Furthermore, 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 is SEQ ID NO: 24 or SEQ ID NO: 58. The amino acid sequence of the TERT antigen linked to the IgE leader may be linked to an HA tag.

[0088] Some embodiments relate to a protein homologous to SEQ ID NO: 24 or SEQ ID NO: 58. Some embodiments relate to an immunogenic protein having 95% homology with the protein sequence set forth in SEQ ID NO: 24 or SEQ ID NO: 58. Some embodiments relate to an immunogenic protein having 96% homology with the protein sequence set forth in SEQ ID NO: 24 or SEQ ID NO: 58. Some embodiments relate to an immunogenic protein having 97% homology with the protein sequence set forth in SEQ ID NO: 24 or SEQ ID NO: 58. Some embodiments relate to an immunogenic protein having 98% homology with the protein sequence set forth in SEQ ID NO: 24 or SEQ ID NO: 58. Some embodiments relate to an immunogenic protein having 99% homology with the protein sequence set forth in SEQ ID NO: 24 or SEQ ID NO: 58.

[0089] Some embodiments relate to a protein having identity with SEQ ID NO: 24 or SEQ ID NO: 58. Some embodiments relate to an immunogenic protein having an amino acid sequence that has 80% identity with the full-length amino acid sequence set forth in SEQ ID NO: 24 or SEQ ID NO: 58. Some embodiments relate to an immunogenic protein having an amino acid sequence that has 85% identity with the full-length amino acid sequence set forth in SEQ ID NO: 24 or SEQ ID NO: 58. Some embodiments relate to an immunogenic protein having an amino acid sequence that has 90% identity with the full-length amino acid sequence set forth in SEQ ID NO: 24 or SEQ ID NO: 58. Some embodiments relate to an immunogenic protein having an amino acid sequence that has 91% identity with the full-length amino acid sequence set forth in SEQ ID NO: 24 or SEQ ID NO: 58. Some embodiments relate to an immunogenic protein having an amino acid sequence that has 92% identity with the full-length amino acid sequence set forth in SEQ ID NO: 24 or SEQ ID NO: 58. Some embodiments relate to an immunogenic protein having an amino acid sequence that has 93% identity with the full-length amino acid sequence set forth in SEQ ID NO: 24 or SEQ ID NO: 58. Some embodiments relate to an immunogenic protein having an amino acid sequence that has 94% identity with the full-length amino acid sequence set forth in SEQ ID NO: 24 or SEQ ID NO: 58. Some embodiments relate to an immunogenic protein having an amino acid sequence that has 95% identity with the full-length amino acid sequence set forth in SEQ ID NO: 24 or SEQ ID NO: 58. Some embodiments relate to an immunogenic protein having an amino acid sequence that has 96% identity with the full-length amino acid sequence set forth in SEQ ID NO: 24 or SEQ ID NO: 58. Some embodiments relate to an immunogenic protein having an amino acid sequence that has 97% identity with the full-length amino acid sequence set forth in SEQ ID NO: 24 or SEQ ID NO: 58. Some embodiments relate to an immunogenic protein having an amino acid sequence that has 98% identity with the full-length amino acid sequence set forth in SEQ ID NO: 24 or SEQ ID NO: 58. Some embodiments relate to an immunogenic protein having an amino acid sequence that has 99% identity with the full-length amino acid sequence set forth in SEQ ID NO: 24 or SEQ ID NO: 58.

[0090] In some embodiments, the protein does not include a leader sequence. In some embodiments, the protein does not include an IgE leader. A fragment of the protein can include 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 can be provided. The immunogenic fragment can include 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 an immunoglobulin leader, such as an IgE leader. In some embodiments, the fragment does not include a leader sequence. In some embodiments, the fragment does not include a leader sequence such as an IgE leader.

[0091] An immunogenic fragment of a protein having an amino acid sequence homologous to the immunogenic fragment of SEQ ID NO: 24 or SEQ ID NO: 58 can be provided. Such an immunogenic fragment 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 more homology to SEQ ID NO: 18. Some embodiments relate to an immunogenic fragment having 96% homology to an immunogenic fragment of the protein sequences herein. Some embodiments relate to an immunogenic fragment having 97% homology to an immunogenic fragment of the protein sequences herein. Some embodiments relate to an immunogenic fragment having 98% homology to an immunogenic fragment of the protein sequences herein. Some embodiments relate to an immunogenic fragment having 99% homology to an immunogenic fragment of the protein sequences herein. In some embodiments, the fragment comprises a leader sequence, such as an immunoglobulin leader, such as an IgE leader. In some embodiments, the fragment does not comprise a leader sequence. In some embodiments, the fragment does not comprise a leader sequence such as an IgE leader.

[0092] It is possible to provide an immunogenic fragment of a protein having an amino acid sequence identical to the immunogenic fragment of SEQ ID NO: 24 or SEQ ID NO: 58. Such an immunogenic fragment 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% identity to the amino acid sequence set forth in SEQ ID NO: 24 or SEQ ID NO: 58. In some embodiments, the fragment comprises a leader sequence, such as an immunoglobulin leader, such as an IgE leader. In some embodiments, the fragment does not comprise a leader sequence. In some embodiments, the fragment does not comprise a leader sequence such as an IgE leader.

[0093] As referred to herein with respect to ligating a signal peptide or leader sequence to the N-terminus of a protein, the signal peptide / leader sequence replaces the N-terminal methionine of the protein encoded by the nucleic acid sequence encoding that protein that does not include the coding sequence of the signal peptide.

[0094] The fragment of SEQ ID NO: 23 or SEQ ID NO: 57 can comprise 30 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, the fragment of SEQ ID NO: 23 or SEQ ID NO: 57 can comprise 45 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, the fragment of SEQ ID NO: 23 or SEQ ID NO: 57 can comprise 60 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, the fragment of SEQ ID NO: 23 or SEQ ID NO: 57 can comprise 75 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, the fragment of SEQ ID NO: 23 or SEQ ID NO: 57 can comprise 90 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, the fragment of SEQ ID NO: 23 or SEQ ID NO: 57 can comprise 120 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, the fragment of SEQ ID NO: 23 or SEQ ID NO: 57 can comprise 150 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, the fragment of SEQ ID NO: 23 or SEQ ID NO: 57 can comprise 180 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, the fragment of SEQ ID NO: 23 or SEQ ID NO: 57 can comprise 210 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, the fragment of SEQ ID NO: 23 or SEQ ID NO: 57 can comprise 240 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, the fragment of SEQ ID NO: 23 or SEQ ID NO: 57 can comprise 270 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, the fragment of SEQ ID NO: 23 or SEQ ID NO: 57 can comprise 300 or more nucleotides, including a sequence encoding an immunodominant epitope.In some embodiments, the fragment of SEQ ID NO: 23 or SEQ ID NO: 57 can contain 360 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, the fragment of SEQ ID NO: 23 or SEQ ID NO: 57 can contain 420 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, the fragment of SEQ ID NO: 23 or SEQ ID NO: 57 can contain 480 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, the fragment of SEQ ID NO: 23 or SEQ ID NO: 57 can contain 540 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, the fragment of SEQ ID NO: 23 or SEQ ID NO: 57 can contain 600 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, the fragment of SEQ ID NO: 23 or SEQ ID NO: 57 can contain 300 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, the fragment of SEQ ID NO: 23 or SEQ ID NO: 57 can contain 660 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, the fragment of SEQ ID NO: 23 or SEQ ID NO: 57 can contain 720 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, the fragment of SEQ ID NO: 23 or SEQ ID NO: 57 can contain 780 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, the fragment of SEQ ID NO: 23 or SEQ ID NO: 57 can contain 840 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, the fragment of SEQ ID NO: 23 or SEQ ID NO: 57 can contain 900 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, the fragment of SEQ ID NO: 23 or SEQ ID NO: 57 can contain 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 can 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 can 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 can 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 can 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 can 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 can 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 can 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 can 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 can 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 can 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 can 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 can comprise 1680 or more nucleotides, including a sequence encoding an immunodominant epitope.In some embodiments, the fragment of SEQ ID NO: 23 or SEQ ID NO: 57 can contain 1740 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, the fragment of SEQ ID NO: 23 or SEQ ID NO: 57 can contain 1800 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, the fragment of SEQ ID NO: 23 or SEQ ID NO: 57 can contain 1860 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, the fragment of SEQ ID NO: 23 or SEQ ID NO: 57 can contain 1920 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, the fragment of SEQ ID NO: 23 or SEQ ID NO: 57 can contain 1980 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, the fragment of SEQ ID NO: 23 or SEQ ID NO: 57 can contain 2040 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, the fragment of SEQ ID NO: 23 or SEQ ID NO: 57 can contain 2100 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, the fragment of SEQ ID NO: 23 or SEQ ID NO: 57 can contain 2160 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, the fragment of SEQ ID NO: 23 or SEQ ID NO: 57 can contain 2220 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, the fragment of SEQ ID NO: 23 or SEQ ID NO: 57 can contain 2280 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, the fragment of SEQ ID NO: 23 or SEQ ID NO: 57 can contain 2340 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, the fragment of SEQ ID NO: 23 or SEQ ID NO: 57 can contain 2400 or more nucleotides, including a sequence encoding an immunodominant epitope.In some embodiments, the fragment of SEQ ID NO: 23 or SEQ ID NO: 57 can comprise 2460 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, the fragment of SEQ ID NO: 23 or SEQ ID NO: 57 can comprise 2520 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, the fragment of SEQ ID NO: 23 or SEQ ID NO: 57 can comprise 2580 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, the fragment of SEQ ID NO: 23 or SEQ ID NO: 57 can comprise 2640 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, the fragment of SEQ ID NO: 23 or SEQ ID NO: 57 can comprise 2700 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, the fragment of SEQ ID NO: 23 or SEQ ID NO: 57 can comprise 2760 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, the fragment of SEQ ID NO: 23 or SEQ ID NO: 57 can comprise 2820 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, the fragment of SEQ ID NO: 23 or SEQ ID NO: 57 can comprise 2880 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, the fragment of SEQ ID NO: 23 or SEQ ID NO: 57 can comprise 2940 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, the fragment of SEQ ID NO: 23 or SEQ ID NO: 57 can comprise 3000 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, the fragment of SEQ ID NO: 23 or SEQ ID NO: 57 can comprise 3060 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, the fragment of SEQ ID NO: 23 or SEQ ID NO: 57 can comprise 3120 or more nucleotides, including a sequence encoding an immunodominant epitope.In some embodiments, the fragment of SEQ ID NO: 23 or SEQ ID NO: 57 can comprise 3180 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, the fragment of SEQ ID NO: 23 or SEQ ID NO: 57 can comprise 3240 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, the fragment of SEQ ID NO: 23 or SEQ ID NO: 57 can comprise 3300 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, the fragment of SEQ ID NO: 23 or SEQ ID NO: 57 can comprise 3 or more nucleotides. In some embodiments, the fragment of SEQ ID NO: 23 or SEQ ID NO: 57 can comprise 3420 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, the fragment of SEQ ID NO: 23 or SEQ ID NO: 57 can comprise 3480 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, the fragment of SEQ ID NO: 23 or SEQ ID NO: 57 can comprise the coding sequence of the IgE leader sequence. In some embodiments, the fragment of SEQ ID NO: 23 or SEQ ID NO: 57 does not comprise the coding sequence of the IgE leader sequence.

[0095] The fragment has fewer than 60 nucleotides, in some embodiments fewer than 75 nucleotides, in some embodiments fewer than 90 nucleotides, in some embodiments fewer than 120 nucleotides, in some embodiments fewer than 150 nucleotides, in some embodiments fewer than 180 nucleotides, in some embodiments fewer than 210 nucleotides, in some embodiments fewer than 240 nucleotides, in some embodiments fewer than 270 nucleotides, in some embodiments fewer than 300 nucleotides, in some embodiments fewer than 360 nucleotides, in some embodiments fewer than 420 nucleotides, in some embodiments fewer than 480 nucleotides, in some embodiments fewer than 540 nucleotides, in some embodiments fewer than 600 nucleotides, in some embodiments fewer than 660 nucleotides, in some embodiments fewer than 720 nucleotides, in some embodiments fewer than 780 nucleotides, in some embodiments fewer than 840 nucleotides, in some embodiments fewer than 900 nucleotides, in some embodiments fewer than 960 nucleotides, in some embodiments fewer than 1020 nucleotides, in some embodiments fewer than 1080 nucleotides, in some embodiments fewer than 1140 nucleotides, in some embodiments fewer than 1200 nucleotides, in some embodiments fewer than 1260 nucleotides, in some embodiments fewer than 1320 nucleotides, in some embodiments fewer than 1380 nucleotides, in some embodiments fewer than 1440 nucleotides, in some embodiments fewer than 1500 nucleotides, in some embodiments fewer than 1560 nucleotides, in some embodiments fewer than 1620 nucleotides, in some embodiments fewer than 1680 nucleotides, in some embodiments fewer than 1740 nucleotides,In some embodiments, it can include fewer than 1800 nucleotides, in some embodiments, fewer than 1860 nucleotides, in some embodiments, fewer than 1920 nucleotides, in some embodiments, fewer than 1980 nucleotides, in some embodiments, fewer than 2040 nucleotides, in some embodiments, fewer than 2100 nucleotides, in some embodiments, fewer than 2160 nucleotides, in some embodiments, fewer than 2220 nucleotides, in some embodiments, fewer than 2280 nucleotides, in some embodiments, fewer than 2340 nucleotides, in some embodiments, fewer than 2400 nucleotides, in some embodiments, fewer than 2460 nucleotides, in some embodiments, fewer than 2520 nucleotides, in some embodiments, fewer than 2580 nucleotides, in some embodiments, fewer than 2640 nucleotides, in some embodiments, fewer than 2700 nucleotides, in some embodiments, fewer than 2760 nucleotides, in some embodiments, fewer than 2820 nucleotides, in some embodiments, fewer than 2860 nucleotides, in some embodiments, fewer than 2940 nucleotides, in some embodiments, fewer than 3000 nucleotides, in some embodiments, fewer than 3060 nucleotides, in some embodiments, fewer than 3120 nucleotides, in some embodiments, fewer than 3180 nucleotides, in some embodiments, fewer than 3240 nucleotides, in some embodiments, fewer than 3300 nucleotides, in some embodiments, fewer than 3360 nucleotides, in some embodiments, fewer than 3420 nucleotides, in some embodiments, fewer than 3480 nucleotides, and in some embodiments, fewer than 3510 nucleotides.

[0096] The fragment of SEQ ID NO: 24 or SEQ ID NO: 58 can comprise 15 or more amino acids, including a sequence encoding an immunodominant epitope. In some embodiments, the fragment of SEQ ID NO: 24 or SEQ ID NO: 58 can comprise 18 or more amino acids, including a sequence encoding an immunodominant epitope. In some embodiments, the fragment of SEQ ID NO: 24 or SEQ ID NO: 58 can comprise 21 or more amino acids, including a sequence encoding an immunodominant epitope. In some embodiments, the fragment of SEQ ID NO: 24 or SEQ ID NO: 58 can comprise 24 or more amino acids, including a sequence encoding an immunodominant epitope. In some embodiments, the fragment of SEQ ID NO: 24 or SEQ ID NO: 58 can comprise 30 or more amino acids, including a sequence encoding an immunodominant epitope. In some embodiments, the fragment of SEQ ID NO: 24 or SEQ ID NO: 58 can comprise 36 or more amino acids, including a sequence encoding an immunodominant epitope. In some embodiments, the fragment of SEQ ID NO: 24 or SEQ ID NO: 58 can comprise 42 or more amino acids, including a sequence encoding an immunodominant epitope. In some embodiments, the fragment of SEQ ID NO: 24 or SEQ ID NO: 58 can comprise 48 or more amino acids, including a sequence encoding an immunodominant epitope. In some embodiments, the fragment of SEQ ID NO: 24 or SEQ ID NO: 58 can comprise 54 or more amino acids, including a sequence encoding an immunodominant epitope. In some embodiments, the fragment of SEQ ID NO: 24 or SEQ ID NO: 58 can comprise 60 or more amino acids, including a sequence encoding an immunodominant epitope. In some embodiments, the fragment of SEQ ID NO: 24 or SEQ ID NO: 58 can comprise 66 or more amino acids, including a sequence encoding an immunodominant epitope. In some embodiments, the fragment of SEQ ID NO: 24 or SEQ ID NO: 58 can comprise 72 or more amino acids, including a sequence encoding an immunodominant epitope. In some embodiments, the fragment of SEQ ID NO: 24 or SEQ ID NO: 58 can comprise 90 or more amino acids, including a sequence encoding an immunodominant epitope.In some embodiments, the fragment of SEQ ID NO: 24 or SEQ ID NO: 58 can contain 120 or more amino acids, including a sequence encoding an immunodominant epitope. In some embodiments, the fragment of SEQ ID NO: 24 or SEQ ID NO: 58 can contain 150 or more amino acids, including a sequence encoding an immunodominant epitope. In some embodiments, the fragment of SEQ ID NO: 24 or SEQ ID NO: 58 can contain 180 or more amino acids, including a sequence encoding an immunodominant epitope. In some embodiments, the fragment of SEQ ID NO: 24 or SEQ ID NO: 58 can contain 210 or more amino acids, including a sequence encoding an immunodominant epitope. In some embodiments, the fragment of SEQ ID NO: 24 or SEQ ID NO: 58 can contain 240 or more amino acids, including a sequence encoding an immunodominant epitope. In some embodiments, the fragment of SEQ ID NO: 24 or SEQ ID NO: 58 can contain 270 or more amino acids, including a sequence encoding an immunodominant epitope. In some embodiments, the fragment of SEQ ID NO: 24 or SEQ ID NO: 58 can contain 300 or more amino acids, including a sequence encoding an immunodominant epitope. In some embodiments, the fragment of SEQ ID NO: 24 or SEQ ID NO: 58 can contain 330 or more amino acids, including a sequence encoding an immunodominant epitope. In some embodiments, the fragment of SEQ ID NO: 24 or SEQ ID NO: 58 can contain 360 or more amino acids, including a sequence encoding an immunodominant epitope. In some embodiments, the fragment of SEQ ID NO: 24 or SEQ ID NO: 58 can contain 390 or more amino acids, including a sequence encoding an immunodominant epitope. In some embodiments, the fragment of SEQ ID NO: 24 or SEQ ID NO: 58 can contain 420 or more amino acids, including a sequence encoding an immunodominant epitope. In some embodiments, the fragment of SEQ ID NO: 24 or SEQ ID NO: 58 can contain 450 or more amino acids, including a sequence encoding an immunodominant epitope.In some embodiments, the fragment of SEQ ID NO: 24 or SEQ ID NO: 58 can contain 480 or more amino acids, including a sequence encoding an immunodominant epitope. In some embodiments, the fragment of SEQ ID NO: 24 or SEQ ID NO: 58 can contain 510 or more amino acids, including a sequence encoding an immunodominant epitope. In some embodiments, the fragment of SEQ ID NO: 24 or SEQ ID NO: 58 can contain 540 or more amino acids, including a sequence encoding an immunodominant epitope. In some embodiments, the fragment of SEQ ID NO: 24 or SEQ ID NO: 58 can contain 570 or more amino acids, including a sequence encoding an immunodominant epitope. In some embodiments, the fragment of SEQ ID NO: 24 or SEQ ID NO: 58 can contain 600 or more amino acids, including a sequence encoding an immunodominant epitope. In some embodiments, the fragment of SEQ ID NO: 24 or SEQ ID NO: 58 can contain 630 or more amino acids, including a sequence encoding an immunodominant epitope. In some embodiments, the fragment of SEQ ID NO: 24 or SEQ ID NO: 58 can contain 660 or more amino acids, including a sequence encoding an immunodominant epitope. In some embodiments, the fragment of SEQ ID NO: 24 or SEQ ID NO: 58 can contain 690 or more amino acids, including a sequence encoding an immunodominant epitope. In some embodiments, the fragment of SEQ ID NO: 24 or SEQ ID NO: 58 can contain 720 or more amino acids, including a sequence encoding an immunodominant epitope. In some embodiments, the fragment of SEQ ID NO: 24 or SEQ ID NO: 58 can contain 750 or more amino acids, including a sequence encoding an immunodominant epitope. In some embodiments, the fragment of SEQ ID NO: 24 or SEQ ID NO: 58 can contain 780 or more amino acids, including a sequence encoding an immunodominant epitope. In some embodiments, the fragment of SEQ ID NO: 24 or SEQ ID NO: 58 can contain 810 or more amino acids, including a sequence encoding an immunodominant epitope.In some embodiments, the fragment of SEQ ID NO: 24 or SEQ ID NO: 58 can contain 840 or more amino acids, including a sequence encoding an immunodominant epitope. In some embodiments, the fragment of SEQ ID NO: 24 or SEQ ID NO: 58 can contain 870 or more amino acids, including a sequence encoding an immunodominant epitope. In some embodiments, the fragment of SEQ ID NO: 24 or SEQ ID NO: 58 can contain 900 or more amino acids, including a sequence encoding an immunodominant epitope. In some embodiments, the fragment of SEQ ID NO: 24 or SEQ ID NO: 58 can contain 930 or more amino acids, including a sequence encoding an immunodominant epitope. In some embodiments, the fragment of SEQ ID NO: 24 or SEQ ID NO: 58 can contain 960 or more amino acids, including a sequence encoding an immunodominant epitope. In some embodiments, the fragment of SEQ ID NO: 24 or SEQ ID NO: 58 can contain 990 or more amino acids, including a sequence encoding an immunodominant epitope. In some embodiments, the fragment of SEQ ID NO: 24 or SEQ ID NO: 58 can contain 1020 or more amino acids, including a sequence encoding an immunodominant epitope. In some embodiments, the fragment of SEQ ID NO: 24 or SEQ ID NO: 58 can contain 1050 or more amino acids, including a sequence encoding an immunodominant epitope. In some embodiments, the fragment of SEQ ID NO: 24 or SEQ ID NO: 58 can contain 1080 or more amino acids, including a sequence encoding an immunodominant epitope. In some embodiments, the fragment of SEQ ID NO: 24 or SEQ ID NO: 58 can contain 1110 or more amino acids, including a sequence encoding an immunodominant epitope. In some embodiments, the fragment of SEQ ID NO: 24 or SEQ ID NO: 58 can contain 1140 or more amino acids, including a sequence encoding an immunodominant epitope. In some embodiments, the fragment of SEQ ID NO: 24 or SEQ ID NO: 58 can contain 1170 or more amino acids, including a sequence encoding an immunodominant epitope.In some embodiments, the fragment of SEQ ID NO: 24 or SEQ ID NO: 58 can contain 1200 or more amino acids, including a sequence encoding an immunodominant epitope. In some embodiments, the fragment of SEQ ID NO: 24 or SEQ ID NO: 58 can contain 1230 or more amino acids, including a sequence encoding an immunodominant epitope. In some embodiments, the fragment of SEQ ID NO: 24 or SEQ ID NO: 58 can contain 1260 or more amino acids, including a sequence encoding an immunodominant epitope. In some embodiments, the fragment of SEQ ID NO: 24 or SEQ ID NO: 58 can contain 1290 or more amino acids, including a sequence encoding an immunodominant epitope. In some embodiments, the fragment of SEQ ID NO: 24 or SEQ ID NO: 58 can contain 1320 or more amino acids, including a sequence encoding an immunodominant epitope. In some embodiments, the fragment of SEQ ID NO: 24 or SEQ ID NO: 58 can contain 1350 or more amino acids, including a sequence encoding an immunodominant epitope. In some embodiments, the fragment of SEQ ID NO: 24 or SEQ ID NO: 58 can contain 1380 or more amino acids, including a sequence encoding an immunodominant epitope. In some embodiments, the fragment of SEQ ID NO: 24 or SEQ ID NO: 58 can contain 1410 or more amino acids, including a sequence encoding an immunodominant epitope. In some embodiments, the fragment of SEQ ID NO: 24 or SEQ ID NO: 58 can contain 1440 or more amino acids, including a sequence encoding an immunodominant epitope. In some embodiments, the fragment of SEQ ID NO: 24 or SEQ ID NO: 58 can contain 1470 or more amino acids, including a sequence encoding an immunodominant epitope. In some embodiments, the fragment of SEQ ID NO: 24 or SEQ ID NO: 58 can contain 1500 or more amino acids, including a sequence encoding an immunodominant epitope. In some embodiments, the fragment of SEQ ID NO: 24 or SEQ ID NO: 58 can contain the coding sequence of the IgE leader sequence. In some embodiments, the fragment of SEQ ID NO: 24 or SEQ ID NO: 58 does not contain the coding sequence of the IgE leader sequence.

[0097] The fragment has fewer than 24 amino acids, in some embodiments fewer than 30 amino acids, in some embodiments fewer than 36 amino acids, in some embodiments fewer than 42 amino acids, in some embodiments fewer than 48 amino acids, in some embodiments fewer than 54 amino acids, in some embodiments fewer than 60 amino acids, in some embodiments fewer than 72 amino acids, in some embodiments fewer than 90 amino acids, in some embodiments fewer than 120 amino acids, in some embodiments fewer than 150 amino acids, in some embodiments fewer than 180 amino acids, in some embodiments fewer than 210 amino acids, in some embodiments fewer than 240 amino acids, in some embodiments fewer than 260 amino acids, in some embodiments fewer than 290 amino acids, in some embodiments fewer than 320 amino acids, in some embodiments fewer than 350 amino acids, in some embodiments fewer than 380 amino acids, in some embodiments fewer than 410 amino acids, in some embodiments fewer than 440 amino acids, in some embodiments fewer than 470 amino acids, in some embodiments fewer than 500 amino acids, in some embodiments fewer than 530 amino acids, in some embodiments fewer than 560 amino acids, in some embodiments fewer than 590 amino acids, in some embodiments fewer than 620 amino acids, in some embodiments fewer than 650 amino acids, in some embodiments fewer than 680 amino acids, in some embodiments fewer than 710 amino acids, in some embodiments fewer than 740 amino acids, in some embodiments fewer than 770 amino acids, in some embodiments fewer than 800 amino acids, in some embodiments fewer than 830 amino acids, in some embodiments fewer than 860 amino acids, in some embodiments fewer than 890 amino acids, in some embodiments fewer than 920 amino acids, in some embodiments fewer thanIn some embodiments, the amino acid sequence may comprise fewer than 950 amino acids, in some embodiments, fewer than 980 amino acids, in some embodiments, fewer than 1010 amino acids, in some embodiments, fewer than 1040 amino acids, in some embodiments, fewer than 1070 amino acids, in some embodiments, fewer than 1200 amino acids, in some embodiments, fewer than 1230 amino acids, in some embodiments, fewer than 1260 amino acids, in some embodiments, fewer than 1290 amino acids, in some embodiments, fewer than 1320 amino acids, in some embodiments, fewer than 1350 amino acids, in some embodiments, fewer than 1380 amino acids, in some embodiments, fewer than 1410 amino acids, in some embodiments, fewer than 1440 amino acids, in some embodiments, fewer than 1470 amino acids, and in some embodiments, fewer than 1500 amino acids.

[0098] In one embodiment, the TERT antigen is a synthetic consensus TERT antigen. In certain embodiments, the synthetic consensus TERT has 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 compared to natural TERT antigen.

[0099] In one embodiment, the consensus hTERT antigen may be the nucleic acid sequence of SEQ ID NO: 45 encoding the amino acid sequence of SEQ ID NO: 46. SEQ ID NO: 45 encodes a 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 may not contain or be linked to an IgE leader sequence and / or an HA tag. In one embodiment, the consensus hTERT antigen may be the nucleic acid sequence of SEQ ID NO: 47 encoding the amino acid sequence of SEQ ID NO: 48. SEQ ID NO: 47 encodes a consensus hTERT antigen that does not contain or is not linked to an IgE leader sequence and / or an HA tag.

[0100] In one embodiment, the consensus mTERT antigen may be the nucleic acid sequence of SEQ ID NO: 49 encoding the amino acid sequence of SEQ ID NO: 50. SEQ ID NO: 49 encodes a 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 may not contain or be linked to an IgE leader sequence and / or an HA tag. In one embodiment, the consensus mTERT antigen may be the nucleic acid sequence of SEQ ID NO: 51 encoding the amino acid sequence of SEQ ID NO: 52. SEQ ID NO: 51 encodes a consensus mTERT antigen that does not contain or is not linked to an IgE leader sequence and / or an HA tag.

[0101] In one embodiment, the consensus rhTERT antigen may be the nucleic acid sequence of SEQ ID NO: 53 encoding the amino acid sequence of SEQ ID NO: 54. SEQ ID NO: 53 encodes a 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 may not contain or be linked to an IgE leader sequence and / or an HA tag. In one embodiment, the consensus rhTERT antigen may be the nucleic acid sequence of SEQ ID NO: 55 encoding the amino acid sequence of SEQ ID NO: 56. SEQ ID NO: 55 encodes a consensus rhTERT antigen that does not contain or is not linked to an IgE leader sequence and / or an HA tag.

[0102] In some embodiments, the consensus TERT antigen is a nucleic acid sequence having 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% identity over the full length of the nucleic acid sequences set forth 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 is a nucleic acid sequence encoding an amino acid sequence having 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% identity over the full length of the amino acid sequences set forth 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 may be an amino acid sequence having 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% identity over the full length of the amino acid sequences set forth 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.

[0103] Some embodiments relate to nucleic acid sequences encoding a protein homologous to the consensus TERT antigen, an immunogenic fragment of the consensus TERT antigen, and an immunogenic fragment of a homologous protein. Such nucleic acids can be provided that encode an immunogenic protein having 95% or less homology to the sequence, 96% or less homology to the sequence, 97% or less homology to the sequence, 98% or less homology to the sequence, and 99% or less homology. Similarly, nucleic acid sequences encoding the immunogenic fragments described herein and immunogenic fragments of proteins homologous to the proteins described herein are also provided.

[0104] Some embodiments relate to nucleic acid molecules encoding an immunogenic protein having 95% homology to the nucleic acid coding sequences herein. Some embodiments relate to nucleic acid molecules encoding an immunogenic protein having 96% homology to the nucleic acid coding sequences herein. Some embodiments relate to nucleic acid molecules encoding an immunogenic protein having 97% homology to the nucleic acid coding sequences herein. Some embodiments relate to nucleic acid molecules encoding an immunogenic protein having 98% homology to the nucleic acid coding sequences herein. Some embodiments relate to nucleic acid molecules encoding an immunogenic protein having 99% homology to the nucleic acid coding sequences herein. In some embodiments, a nucleic acid molecule having a coding sequence homologous to the coding sequence of the consensus protein disclosed herein includes a sequence encoding an IgE leader sequence linked to the 5' end of the coding sequence encoding the homologous protein sequence disclosed herein.

[0105] Some embodiments relate to nucleic acid sequences encoding a protein having a specific identity (%) to the full-length consensus TERT antigen, an immunogenic fragment of the consensus TERT antigen, and an immunogenic fragment of a protein having identity to the consensus TERT antigen. Such nucleic acid molecules can be provided that encode immunogenic proteins having 80% or less identity to the full-length consensus TERT sequence, 85% or less identity to the full-length consensus TERT sequence, 90% or less identity to the full-length consensus TERT sequence, 91% or less identity to the full-length consensus TERT sequence, 92% or less identity to the full-length consensus TERT sequence, 93% or less identity to the full-length consensus TERT sequence, 94% or less identity to the full-length consensus TERT sequence, 95% or less identity to the full-length consensus TERT sequence, 96% or less identity to the full-length consensus TERT sequence, 97% or less identity to the full-length consensus TERT sequence, 98% or less identity to the full-length consensus TERT sequence, and 99% or less identity to the full-length consensus TERT sequence. Similarly, provided are nucleic acid sequences encoding the immunogenic fragments described herein and immunogenic fragments of proteins having the same identity (%) as the consensus TERT antigen described herein as shown above.

[0106] In some embodiments, the nucleic acid sequence does not include a coding sequence encoding a leader sequence. In some embodiments, the nucleic acid sequence does not include a coding sequence encoding an IgE leader.

[0107] 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 may 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. The fragment may 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. 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: 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 fragment comprises a coding sequence encoding a leader sequence, such as an immunoglobulin leader, such as an IgE leader. In some embodiments, the fragment does not comprise a coding sequence encoding a leader sequence. In some embodiments, the fragment does not comprise a coding sequence encoding a leader sequence such as an IgE leader.

[0108] Furthermore, 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 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 may be linked to the HA tag.

[0109] Some embodiments relate to a protein 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: 5). Some embodiments relate to an immunogenic protein having 95% homology with the protein sequence set forth 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 an immunogenic protein having 96% homology with the protein sequence set forth 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 an immunogenic protein having 97% homology with the protein sequence set forth 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 an immunogenic protein having 98% homology with the protein sequence set forth in SEQ ID NO: 46. Some embodiments relate to an immunogenic protein having 99% homology with the protein sequence set forth 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.

[0110] Some embodiments relate to a protein 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 an immunogenic protein having an amino acid sequence with 80% identity to the full-length amino acid sequence set forth 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 an immunogenic protein having an amino acid sequence with 85% identity to the full-length amino acid sequence set forth 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 an immunogenic protein having an amino acid sequence with 90% identity to the full-length amino acid sequence set forth 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 an immunogenic protein having an amino acid sequence with 91% identity to the full-length amino acid sequence set forth 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 an immunogenic protein having an amino acid sequence with 92% identity to the full-length amino acid sequence set forth 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 an immunogenic protein having an amino acid sequence with 93% identity to the full-length amino acid sequence set forth 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 an immunogenic protein having an amino acid sequence with 94% identity to the full-length amino acid sequence set forth 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 an immunogenic protein having an amino acid sequence with 95% identity to the full-length amino acid sequence set forth 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 an immunogenic protein having an amino acid sequence that has 96% identity with the full-length amino acid sequence set forth 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 an immunogenic protein having an amino acid sequence that has 97% identity with the full-length amino acid sequence set forth 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 an immunogenic protein having an amino acid sequence that has 98% identity with the full-length amino acid sequence set forth 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 an immunogenic protein having an amino acid sequence that has 99% identity with the full-length amino acid sequence set forth 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.

[0111] In some embodiments, the protein does not include a leader sequence. In some embodiments, the protein does not include an IgE leader. Fragments of the protein can include 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 can include 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 an immunoglobulin leader, such as an IgE leader. In some embodiments, the fragment does not include a leader sequence. In some embodiments, the fragment does not include a leader sequence such as an IgE leader.

[0112] An immunogenic fragment of a protein having an amino acid sequence homologous to the 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 can be provided. Such an immunogenic fragment 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 more homology with 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 an immunogenic fragment having 96% homology with the immunogenic fragment of the protein sequences herein. Some embodiments relate to an immunogenic fragment having 97% homology with the immunogenic fragment of the protein sequences herein. Some embodiments relate to an immunogenic fragment having 98% homology with the immunogenic fragment of the protein sequences herein. Some embodiments relate to an immunogenic fragment having 99% homology with the immunogenic fragment of the protein sequences herein. In some embodiments, the fragment comprises a leader sequence, such as an immunoglobulin leader, such as an IgE leader. In some embodiments, the fragment does not comprise a leader sequence. In some embodiments, the fragment does not comprise a leader sequence such as an IgE leader.

[0113] An immunogenic fragment of a protein having an amino acid sequence having identity with the 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 can be provided. Such an immunogenic fragment has 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% identity with the amino acid sequence set forth 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 an immunoglobulin leader, such as an IgE leader. In some embodiments, the fragment does not include a leader sequence. In some embodiments, the fragment does not include a leader sequence such as an IgE leader.

[0114] When referring herein to ligating a signal peptide or leader sequence to the N-terminus of a protein, the signal peptide / leader sequence replaces the methionine at the N-terminus of the protein encoded by the start codon of the nucleic acid sequence encoding that protein, which does not include the coding sequence of the signal peptide.

[0115] 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 can contain 30 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, 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 can contain 45 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, 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 can contain 60 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, 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 can contain 75 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, 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 can contain 90 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, 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 can contain 120 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, 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 can contain 150 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, 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 can contain 180 or more nucleotides, including a sequence encoding an immunodominant epitope. In some embodiments, 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 can contain 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 can 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 can 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 can 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 can 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 can 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 can 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 can 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 can 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 can 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 can contain 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 can contain 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 can contain 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 can contain 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 can contain 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 can contain 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 can contain 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 can contain 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 can contain 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 can 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 can 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 can 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 can 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 can 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 can 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 can 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 can 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 can 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 can 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 can 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 can 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 can 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 can 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 can 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 can 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 can 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 can 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 can comprise 2,280 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 comprises 2,340 or more nucleotides, including a sequence encoding an immunodominant epitope. It can include. 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 can include 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 can include 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 can include 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 can include 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 can include 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 can include 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 can include 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 can include 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 can include 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 can 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 can 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 can 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 can 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 can 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 can 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 can 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 can 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 can 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 can comprise 3,480 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 can comprise a coding sequence of an IgE leader sequence. 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 does not comprise a coding sequence of an IgE leader sequence.

[0116] The fragment has fewer than 60 nucleotides, in some embodiments fewer than 75 nucleotides, in some embodiments fewer than 90 nucleotides, in some embodiments fewer than 120 nucleotides, in some embodiments fewer than 150 nucleotides, in some embodiments fewer than 180 nucleotides, in some embodiments fewer than 210 nucleotides, in some embodiments fewer than 240 nucleotides, in some embodiments fewer than 270 nucleotides, in some embodiments fewer than 300 nucleotides, in some embodiments fewer than 360 nucleotides, in some embodiments fewer than 420 nucleotides, in some embodiments fewer than 480 nucleotides, in some embodiments fewer than 540 nucleotides, in some embodiments fewer than 600 nucleotides, in some embodiments fewer than 660 nucleotides, in some embodiments fewer than 720 nucleotides, in some embodiments fewer than 780 nucleotides, in some embodiments fewer than 840 nucleotides, in some embodiments fewer than 900 nucleotides, in some embodiments fewer than 960 nucleotides, in some embodiments fewer than 1020 nucleotides, in some embodiments fewer than 1080 nucleotides, in some embodiments fewer than 1140 nucleotides, in some embodiments fewer than 1200 nucleotides, in some embodiments fewer than 1260 nucleotides, in some embodiments fewer than 1320 nucleotides, in some embodiments fewer than 1380 nucleotides, in some embodiments fewer than 1440 nucleotides, in some embodiments fewer than 1500 nucleotides, in some embodiments fewer than 1560 nucleotides, in some embodiments fewer than 1620 nucleotides, in some embodiments fewer than 1680 nucleotides, in some embodiments fewer than 1740 nucleotides,In some embodiments, it can include fewer than 1800 nucleotides, in some embodiments, fewer than 1860 nucleotides, in some embodiments, fewer than 1920 nucleotides, in some embodiments, fewer than 1980 nucleotides, in some embodiments, fewer than 2040 nucleotides, in some embodiments, fewer than 2100 nucleotides, in some embodiments, fewer than 2160 nucleotides, in some embodiments, fewer than 2220 nucleotides, in some embodiments, fewer than 2280 nucleotides, in some embodiments, fewer than 2340 nucleotides, in some embodiments, fewer than 2400 nucleotides, in some embodiments, fewer than 2460 nucleotides, in some embodiments, fewer than 2520 nucleotides, in some embodiments, fewer than 2580 nucleotides, in some embodiments, fewer than 2640 nucleotides, in some embodiments, fewer than 2700 nucleotides, in some embodiments, fewer than 2760 nucleotides, in some embodiments, fewer than 2820 nucleotides, in some embodiments, fewer than 2860 nucleotides, in some embodiments, fewer than 2940 nucleotides, in some embodiments, fewer than 3000 nucleotides, in some embodiments, fewer than 3060 nucleotides, in some embodiments, fewer than 3120 nucleotides, in some embodiments, fewer than 3180 nucleotides, in some embodiments, fewer than 3240 nucleotides, in some embodiments, fewer than 3300 nucleotides, in some embodiments, fewer than 3360 nucleotides, in some embodiments, fewer than 3420 nucleotides, in some embodiments, fewer than 3480 nucleotides, and in some embodiments, fewer than 3510 nucleotides.,

[0117] 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 comprise 15 or more amino acids, including a sequence that comprises an immunodominant epitope. 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 can comprise 18 or more amino acids, including a sequence that comprises an immunodominant epitope. 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 can comprise 21 or more amino acids, including a sequence that comprises an immunodominant epitope. 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 can comprise 24 or more amino acids, including a sequence that comprises an immunodominant epitope. 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 can comprise 30 or more amino acids, including a sequence that comprises an immunodominant epitope. 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 can comprise 36 or more amino acids, including a sequence that comprises an immunodominant epitope. 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 can comprise 42 or more amino acids, including a sequence that comprises an immunodominant epitope. 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 can comprise 48 or more amino acids, including a sequence that comprises an immunodominant epitope. 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 can comprise 54 or more amino acids, including a sequence that comprises an immunodominant epitope.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 can comprise 60 or more amino acids, including a sequence that comprises an immunodominant epitope. 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 can comprise 66 or more amino acids, including a sequence that comprises an immunodominant epitope. 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 can comprise 72 or more amino acids, including a sequence that comprises an immunodominant epitope. 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 can comprise 90 or more amino acids, including a sequence that comprises an immunodominant epitope. 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 can comprise 120 or more amino acids, including a sequence that comprises an immunodominant epitope. 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 can comprise 150 or more amino acids, including a sequence that comprises an immunodominant epitope. 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 can comprise 180 or more amino acids, including a sequence that comprises an immunodominant epitope. 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 can comprise 210 or more amino acids, including a sequence that comprises an immunodominant epitope. 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 can comprise 240 or more amino acids, including a sequence that comprises an immunodominant epitope.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 can comprise 270 or more amino acids, including a sequence comprising the immunodominant epitope. 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 can comprise 300 or more amino acids, including a sequence comprising the immunodominant epitope. 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 can comprise 330 or more amino acids, including a sequence comprising the immunodominant epitope. 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 can comprise 360 or more amino acids, including a sequence comprising the immunodominant epitope. 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 can comprise 390 or more amino acids, including a sequence comprising the immunodominant epitope. 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 can comprise 420 or more amino acids, including a sequence comprising the immunodominant epitope. 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 can comprise 450 or more amino acids, including a sequence comprising the immunodominant epitope. 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 can comprise 480 or more amino acids, including a sequence comprising the immunodominant epitope. 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 can contain 510 or more amino acids, including a sequence that includes an immunodominant epitope.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 can comprise 540 or more amino acids, comprising a sequence that includes an immunodominant epitope. 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 can comprise 570 or more amino acids, comprising a sequence that includes an immunodominant epitope. 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 can comprise 600 or more amino acids, comprising a sequence that includes an immunodominant epitope. 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 can comprise 630 or more amino acids, comprising a sequence that includes an immunodominant epitope. 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 can comprise 660 or more amino acids, comprising a sequence that includes an immunodominant epitope. 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 can comprise 690 or more amino acids, comprising a sequence that includes an immunodominant epitope. 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 can comprise 720 or more amino acids, comprising a sequence that includes an immunodominant epitope. 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 can comprise 750 or more amino acids, comprising a sequence that includes an immunodominant epitope. 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 can comprise 780 or more amino acids, comprising a sequence that includes an immunodominant epitope.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 can comprise 810 or more amino acids, including a sequence that comprises an immunodominant epitope. 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 can comprise 840 or more amino acids, including a sequence that comprises an immunodominant epitope. 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 can comprise 870 or more amino acids, including a sequence that comprises an immunodominant epitope. 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 can comprise 900 or more amino acids, including a sequence that comprises an immunodominant epitope. 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 can comprise 930 or more amino acids, including a sequence that comprises an immunodominant epitope. 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 can comprise 960 or more amino acids, including a sequence that comprises an immunodominant epitope. 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 can comprise 990 or more amino acids, including a sequence that comprises an immunodominant epitope. 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 can comprise 1020 or more amino acids, including a sequence that comprises an immunodominant epitope. 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 can comprise 1050 or more amino acids, including a sequence that comprises an immunodominant epitope.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 can comprise 1080 or more amino acids, comprising a sequence that includes an immunodominant epitope. 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 can comprise 1110 or more amino acids, comprising a sequence that includes an immunodominant epitope. 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 can comprise 1140 or more amino acids, comprising a sequence that includes an immunodominant epitope. 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 can comprise 1170 or more amino acids, comprising a sequence that includes an immunodominant epitope. In some embodiments, SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, SEQ ID NO:. 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 can comprise 1200 or more amino acids, including a sequence comprising the immunodominant epitope. 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 can comprise 1230 or more amino acids, including a sequence comprising the immunodominant epitope. 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 can comprise 1260 or more amino acids, including a sequence comprising the immunodominant epitope. 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 can comprise 1290 or more amino acids, including a sequence comprising the immunodominant epitope. 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 can comprise 1320 or more amino acids, including a sequence comprising the immunodominant epitope. 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 can comprise 1350 or more amino acids, including a sequence comprising the immunodominant epitope. 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 can comprise 1380 or more amino acids, including a sequence comprising the immunodominant epitope. 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 can comprise 1410 or more amino acids, including a sequence comprising the immunodominant epitope. 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 can comprise 1440 or more amino acids, including a sequence comprising the immunodominant epitope.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 can comprise 1470 or more amino acids, including a sequence comprising an immunodominant epitope. 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 can comprise 1500 or more amino acids, including a sequence comprising an immunodominant epitope. 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 can comprise a coding sequence for an IgE leader sequence. 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 does not comprise a coding sequence for an IgE leader sequence.

[0118] The fragment has fewer than 24 amino acids, in some embodiments fewer than 30 amino acids, in some embodiments fewer than 36 amino acids, in some embodiments fewer than 42 amino acids, in some embodiments fewer than 48 amino acids, in some embodiments fewer than 54 amino acids, in some embodiments fewer than 60 amino acids, in some embodiments fewer than 72 amino acids, in some embodiments fewer than 90 amino acids, in some embodiments fewer than 120 amino acids, in some embodiments fewer than 150 amino acids, in some embodiments fewer than 180 amino acids, in some embodiments fewer than 210 amino acids, in some embodiments fewer than 240 amino acids, in some embodiments fewer than 260 amino acids, in some embodiments fewer than 290 amino acids, in some embodiments fewer than 320 amino acids, in some embodiments fewer than 350 amino acids, in some embodiments fewer than 380 amino acids, in some embodiments fewer than 410 amino acids, in some embodiments fewer than 440 amino acids, in some embodiments fewer than 470 amino acids, in some embodiments fewer than 500 amino acids, in some embodiments fewer than 530 amino acids, in some embodiments fewer than 560 amino acids, in some embodiments fewer than 590 amino acids, in some embodiments fewer than 620 amino acids, in some embodiments fewer than 650 amino acids, in some embodiments fewer than 680 amino acids, in some embodiments fewer than 710 amino acids, in some embodiments fewer than 740 amino acids, in some embodiments fewer than 770 amino acids, in some embodiments fewer than 800 amino acids, in some embodiments fewer than 830 amino acids, in some embodiments fewer than 860 amino acids, in some embodiments fewer than 890 amino acids, in some embodiments fewer than 920 amino acids, in some embodiments fewer thanIn some embodiments, the amino acid sequence may comprise fewer than 950 amino acids, in some embodiments, fewer than 980 amino acids, in some embodiments, fewer than 1010 amino acids, in some embodiments, fewer than 1040 amino acids, in some embodiments, fewer than 1070 amino acids, in some embodiments, fewer than 1200 amino acids, in some embodiments, fewer than 1230 amino acids, in some embodiments, fewer than 1260 amino acids, in some embodiments, fewer than 1290 amino acids, in some embodiments, fewer than 1320 amino acids, in some embodiments, fewer than 1350 amino acids, in some embodiments, fewer than 1380 amino acids, in some embodiments, fewer than 1410 amino acids, in some embodiments, fewer than 1440 amino acids, in some embodiments, fewer than 1470 amino acids, and in some embodiments, fewer than 1500 amino acids.

[0119] The present invention relates to an anti-cancer immunogenic composition. The immunogenic composition may comprise one or more cancer antigens. The immunogenic composition can prevent tumor growth. The immunogenic composition can reduce tumor growth. The immunogenic composition can prevent tumor cell metastasis. Depending on the cancer antigen, the immunogenic composition can be intended to treat, but is 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.

[0120] The first step in developing the immunogenic composition is to identify cancer antigens that are not recognized by the immune system and are self-antigens. The identified cancer antigens are then modified from self-antigens to foreign antigens so that they can be recognized by the immune system. Redesigning the nucleic acid and amino acid sequences of recombinant cancer antigens from self-antigens to foreign antigens allows the immune system to exceed its tolerance range. To exceed this tolerance range, several redesign strategies, such as those described below, can be applied to cancer antigens.

[0121] The recombinant cancer antigen of the immunogenic composition is not recognized as a self-antigen and thus exceeds the tolerance range. Deviation from the tolerance range induces a response of antigen-specific T cells and / or high-titer antibodies, thereby inducing or eliciting an immune response directed against or reactive to the 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-gamma (IFN-γ) and / or tumor necrosis factor alpha (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 the tumor or cancer expressing the antigen, including, but not limited to, factors that downregulate presentation by MHC, antigen-specific regulatory T cells (Tregs), cytokines such as PD-L1, FasL, IL-10, and TGF-β, tumor-associated macrophages, tumor-associated fibroblasts, soluble factors produced by immunosuppressive cells, factors that upregulate CTLA-4, PD-1, MDSC, MCP-1, and immune checkpoint molecules.

[0122] In certain embodiments, the immunogenic composition can mediate tumor elimination or prevent the growth of tumor cells by inducing the following. That is, (1) a humoral immune response via B cells that blocks the production of monocyte chemoattractant protein-1 (MCP-1), thereby inhibiting myeloid derived suppressor cells (MDSC) and suppressing tumor growth, and (2) CD8 to attack and kill tumor cells +such as an increase in cytotoxic T lymphocytes (CTLs), (3) an increase in helper T cell responses, and (4) an increase in inflammatory responses mediated by IFN-γ and TFN-α, or a combination of the above items. The above immunogenic composition can increase the tumor-free survival rate by 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%, 41%, 42%, 43%, 44%, and 45%. The above immunogenic composition can reduce the tumor size 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 above immunogenic composition can prevent and inhibit an increase in monocyte chemoattractant protein-1 (MCP-1), a cytokine secreted by myeloid-derived suppressor cells. The above immunogenic composition can increase the tumor survival rate 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%.

[0123] The above immunogenic composition can increase the cellular immune response in a subject administered with 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 as compared to the cellular immune response in a subject not administered with the immunogenic composition. In some embodiments, the above immunogenic composition can increase the cellular immune response in a subject administered with the immunogenic composition 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, 4500-fold, 4600-fold, 4700-fold, 4800-fold, 4900-fold, 5000-fold, 5100-fold, 5200-fold, 5300-fold, 5400-fold, 5500-fold, 5600-fold, 5700-fold, 5800-fold, 5900-fold, or 6000-fold as compared to the cellular immune response in a subject not administered with the immunogenic composition.

[0124] The above immunogenic composition can increase the interferon-gamma (IFN-γ) concentration in a subject administered with 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 as compared to the IFN-γ concentration in a subject not administered with the immunogenic composition. In some embodiments, the above immunogenic composition can increase the IFN-γ concentration in a subject administered with the immunogenic composition 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, 4500-fold, 4600-fold, 4700-fold, 4800-fold, 4900-fold, 5000-fold, 5100-fold, 5200-fold, 5300-fold, 5400-fold, 5500-fold, 5600-fold, 5700-fold, 5800-fold, 5900-fold, or 6000-fold as compared to the IFN-γ concentration in a subject not administered with the immunogenic composition.

[0125] The above immunogenic composition can be a nucleic acid vaccine. In one embodiment, the nucleic acid vaccine is a DNA vaccine. The 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 hereby incorporated by reference in their entirety. The above DNA vaccine can further contain an element or reagent that inhibits the integration of the vaccine into the chromosome.

[0126] The above immunogenic composition can be an RNA vaccine. The RNA vaccine can be introduced into cells. The RNA vaccine can further contain an element or reagent that inhibits the integration of the RNA vaccine into the chromosome.

[0127] The above immunogenic composition can be, but is not limited to, for example, live attenuated vaccines, vaccines using recombinant vectors for delivering antigens, subunit vaccines, and glycoprotein vaccines such as those described in 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, 5,242,829, 5,294,441, 5,294,548, 5,310,668, 5,387,744, 5,389,368, 5,424,065, 5,451,499, 5,453,364, 5,462,734, 5,470,734, 5,474,935, 5,482,713, 5,591,439, 5,643,579, 5,650,309, 5,698,202, 5,955,088, 6,034,298, 6,042,836, 6,156,319, and 6,589,529, and the above patents are hereby incorporated by reference in their entirety.

[0128] The immunogenic composition of the present invention can have the characteristics required of an effective vaccine, such as being safe so that the vaccine itself does not cause disease or death; being protective against disease; inducing neutralizing antibodies; inducing a defensive response of T cells; and being easy to administer, having few side effects, being biologically stable, and having a low cost per dose. By containing the cancer antigens discussed below, the above immunogenic composition can achieve some or all of these characteristics.

[0129] As will be described in more detail below, the above immunogenic composition can further comprise one or more inhibitors of one or more immune checkpoint molecules (i.e., immune checkpoint inhibitors). Immune checkpoint molecules will be described in more detail below. An immune checkpoint inhibitor is any nucleic acid or protein that prevents the suppression of any element in the immune system, such as presentation by MHC class, presentation and / or differentiation by T cells, presentation and / or differentiation by B cells, any cytokine, chemokine, or signaling for the proliferation and / or differentiation of immune cells. Also as will be described in more detail below, the above immunogenic composition may further be combined with antibodies against checkpoint inhibitors such as PD-1, PDL-1, CTLA4, TIM3, and / or LAG3 to increase the stimulation of both cellular and humoral immune responses. By using antibodies targeting immune checkpoint proteins, it is prevented that the immune checkpoint proteins suppress the responses of T cells and / or B cells.

[0130] An immunogenic composition can also include an antigen, or a fragment or variant thereof. The antigen can be anything that induces an immune response in a subject. The antigen can be a nucleic acid sequence, an amino acid sequence, or a combination thereof. The nucleic acid sequence can be DNA, RNA, cDNA, variants thereof, fragments thereof, or combinations thereof. The nucleic acid can also include a further sequence encoding a linker or tag sequence linked to the antigen by a peptide bond. The amino acid sequence can be a protein, a peptide, variants thereof, fragments thereof, or combinations thereof.

[0131] The antigen can be included, for example, in proteins, nucleic acids, or fragments thereof, or variants thereof, or combinations thereof, derived from any number of organisms such as viruses, parasites, bacteria, fungi, or mammals. The antigen can also be related to autoimmune diseases, allergies, or asthma. In other embodiments, the antigen can be related to cancer, herpes, influenza, hepatitis B, hepatitis C, human papillomavirus (HPV), or human immunodeficiency virus (HIV).

[0132] Some antigens can induce a strong immune response. Other antigens can induce a weak immune response. The antigen can induce a stronger immune response when combined with the TERT antigen.

[0133] a. Cancer antigen The above immunogenic composition can contain one or more cancer antigens. The cancer antigen can be a nucleic acid sequence, an amino acid sequence, or a combination thereof. The above nucleic acid sequence can be DNA, RNA, cDNA, their variants, their fragments, or a combination thereof. The above nucleic acid sequence can also contain a further sequence encoding a linker or tag sequence linked to the cancer antigen by a peptide bond. The above amino acid sequence can be a protein, a peptide, their variants, their fragments, or a combination thereof. The cancer antigen can be a recombinant cancer antigen.

[0134] In the context of the present invention, "tumor antigen" or "hyperproliferative disease antigen" or "antigen associated with a hyperproliferative disease" refers to an antigen common to a specific hyperproliferative disease such as cancer. The antigens considered herein are merely examples. This list is not intended to be exclusive, and additional examples will be immediately apparent to those skilled in the art.

[0135] A cancer antigen, or tumor antigen, is a protein produced by tumor cells that induces an immune response, particularly a T cell-mediated immune response. The selection of the antigen-binding portion of the present invention is determined according to the specific 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, α-fetoprotein (AFP), lectin-reactive AFP, thyroglobulin, RAGE-1, MN-CA IX, human telomerase reverse transcriptase, RU1, RU2(AS), intestinal carboxylesterase, mut hsp70-2, M-CSF, prostase, prostate-specific antigen (PSA), PAP, NY-ESO-1, LAGE-1a, p53, prostain, PSMA, Her2 / neu, survivin and telomerase, prostate cancer tumor antigen-1 (PCTA-1), MAGE, ELF2M, neutrophil elastase, ephrin B2, CD22, insulin growth factor (IGF)-I, IGF-II, IGF-I receptor and mesothelin.

[0136] In one embodiment, the tumor antigen comprises one or more antigenic cancer epitopes associated with malignant tumors. Malignant tumors express many proteins that can serve as target antigens for immune attack. These molecules include, but are not limited to, tissue-specific antigens such as MART-1, tyrosinase, and GP100 in melanoma, and prostatic acid phosphatase (PAP) and prostate-specific antigen (PSA) in prostate cancer. Other target molecules belong to the group of transformation-associated molecules, such as the oncogene HER-2 / Neu / ErbB-2. Yet another group of target antigens is carcinoembryonic antigen (CEA), such as oncofetal antigen. In B-cell lymphomas, tumor-specific idiotypic immunoglobulins constitute truly tumor-specific immunoglobulin antigens unique to individual tumors. B-cell differentiation antigens, such as CD19, CD20, and CD37, are other candidate target antigens in B-cell lymphomas. Some of these antigens (CEA, HER-2, CD19, CD20, idiotypes) have been used as targets for passive immunotherapy using monoclonal antibodies, but with limited success.

[0137] The type of tumor antigen referred to in the present invention may be a tumor-specific antigen (TSA) or a tumor-associated antigen (TAA). TSAs are unique to tumor cells and do not occur in other cells in the body. TAA-associated antigens are not unique to tumor cells and are also expressed in normal cells under conditions that cannot induce a state of immune tolerance to the antigen. Antigen expression in tumors can occur under conditions that allow the immune system to respond to the antigen. TAA may be an antigen expressed in normal cells during fetal development, when the immune system is immature and unable to respond, or it may be an antigen that is normally present at very low levels in normal cells but expressed at significantly higher levels in tumor cells.

[0138] Non-limiting examples of TSA antigens 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-family antigens such as MAGE-1, MAGE-3, BAGE, GAGE-1, GAGE-2, p15; fetal antigens overexpressed such as CEA; tumor genes overexpressed and mutated tumor suppressor genes such as p53, Ras, HER-2 / neu; unique tumor antigens generated by chromosomal translocation; BCR-ABL, E2A-PRL, H4-RET, IGH-IGK, MYL-RAR, etc.; 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, CA19-9, CA72-4, CAM17.1, NuMa, K-ras, β-catenin, CDK4, Mum-1, p15, p16, 43-9F, 5T4, 791Tgp72, alpha-fetoprotein, β-HCG, BCA225, BTAA, CA125, CA15-3\CA27.29\BCAA, CA195, CA242, 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.

[0139] One or more antibodies targeting the TERT antigen and optionally one or more immune checkpoint proteins can be associated with or combined with a tumor antigen, or a fragment or variant thereof. A cancer marker is a known protein that is present in or upregulated in specific cancer cells. A cancer vaccine can be produced by a method of generating an antigen representative of such a marker that breaks tolerance to self. Such a cancer vaccine can include the 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.

[0140] (1) Tyrosinase (Tyr) The immunogenic composition of the present invention can include the cancer antigen tyrosinase (Tyr), a fragment thereof, or a variant thereof. Tyrosinase is a copper-containing enzyme having tyrosine hydroxylase and dopa oxidase catalytic activities that can be found in microorganisms, plants, and animal tissues. Specifically, tyrosinase catalyzes the production of melanin or other pigments by the oxidation of phenols such as tyrosine. Mutations in the TYR gene result in oculocutaneous albinism in mammals, and non-pathological gene polymorphisms in the TYR gene contribute to changes in skin pigmentation.

[0141] Furthermore, in cancers or tumors such as melanoma, tyrosinase can become uncontrolled, leading to an increase in melanin synthesis. Thus, tyrosinase can be a cancer antigen associated with melanoma. In subjects suffering from melanoma, tyrosinase can be a target for cytotoxic T cell recognition. However, in some cases, the immune response against cancers or tumors (including melanoma) is suppressed, resulting in a microenvironment that supports tumor formation and / or growth, and thus can lead to disease progression.

[0142] Immunosuppression can be promoted by myeloid-derived suppressor cells (MDSC), which are a heterogeneous population of immature macrophages, granulocytes, dendritic cells, and myeloid cells. Myeloid cells can be a heterogeneous population of myeloid progenitor cells and immature myeloid cells (IMC). Markers of MDSC include the expression of Gr-1 and CD11b (i.e., Gr-1 + and CD11b + cells).

[0143] The circulation of MDSC is increased due to chronic infection, and the expansion of MDSC numbers can be associated with autoimmunity and inflammation. In particular, the proliferation of MDSC (presence in tumor or cancerous tissue) promotes tumor growth and avoidance of and / or immune modulation from immune detection, and thus, MDSC can affect the immune response to anti-cancer vaccines.

[0144] MDSC is regulated by Regulator of G-protein signaling 2 (Rgs2), which can be highly expressed in tumor-derived MDSC. Rgs2 can also be widely expressed in various cells such as myeloid cells, for example. MDSC from mice with tumors can exhibit different effects compared to MDSC from mice without tumors. One such difference can be the upregulation of chemokine MCP-1 production, which is secreted by MDSC. MCP-1 can promote cell migration through signal transduction via CCR2, a G-protein coupled receptor (GPCR) present on monocytes, endothelial cells, and T cells. Thus, MCP-1 can cause the migration of endothelial cells and thereby promote angiogenesis. Blocking of MCP-1 via neutralizing antibodies can inhibit angiogenesis and thus lead to a reduction in tumor metastasis and an increase in survival rate. Thus, MCP-1 can be considered an angiogenesis factor. In addition to secreting MCP-1, MDSC secrete growth factors and thereby can further contribute to tumor growth.

[0145] The Tyr antigen can induce antigen-specific T cells and / or high-titer antibody responses, thereby inducing or eliciting an immune response directed against or reactive to a cancer or tumor expressing said 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 immune response can include the induction or secretion of interferon-gamma (IFN-γ) and / or tumor necrosis factor alpha (TNF-α). In other embodiments, the induced or elicited immune response can include, but is not limited to, factors that downregulate presentation by MHC, antigen-specific regulatory T cells (Tregs), cytokines such as PD-L1, FasL, IL-10, and TGF-β, tumor-associated macrophages, tumor-associated fibroblasts, soluble factors produced by immunosuppressive cells, and factors that upregulate immune checkpoint molecules such as CTLA-4, PD-1, MDSC, MCP-1, and the like, one or more immunosuppressive factors that promote the growth of a tumor or cancer expressing said antigen can be reduced or inhibited, and the immune checkpoint molecules will be described in more detail later.

[0146] As demonstrated herein, the Tyr antigen induces antigen-specific T cells and high-titer antibody responses against cancer cells or tumor cells (e.g., melanoma cells). Specifically, the Tyr antigen (1) blocks the production of monocyte chemoattractant protein-1 (MCP-1), thereby inhibiting myeloid-derived suppressor cells (MDSC) and generating antibodies that suppress tumor growth through a humoral immune response mediated by B cell responses, (2) attacks and kills tumor cells for CD8 +It is an important target for immune-mediated tumor elimination by inducing an increase in cytotoxic T lymphocytes (CTLs) such as, (3) an increase in helper T cell responses, and (4) an increase in inflammatory responses mediated by IFN-γ and TFN-α, or a combination of the above items. Thus, an immunogenic composition containing a Tyr antigen (e.g., a consensus Tyr antigen, which will be described in more detail later) prevents immunosuppression by reducing the number of MDSCs present in cancer or tumor tissue, and blocks angiogenesis of cancer or tumor tissue by reducing the production or secretion of MCP-1. Therefore, these immunogenic compositions provide a protective immune response against tumor formation and tumor growth. Accordingly, any user can design the immunogenic composition of the present invention to contain a Tyr antigen that provides broad immunity against tumor formation, tumor metastasis, and tumor growth.

[0147] The Tyr antigen can contain protein epitopes that make the antigen particularly effective as an immunogen capable of inducing an anti-Tyr immune response against it. The Tyr antigen can include the full-length translation product, variants thereof, fragments thereof, or combinations thereof. The Tyr antigen can include a consensus protein.

[0148] The nucleic acid sequence encoding the consensus Tyr antigen can be optimized with respect to codon usage frequency and the corresponding RNA transcript. 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 improve the efficiency of translation. The nucleic acid encoding the consensus Tyr antigen can include multiple stop codons (e.g., TGA TGA) to improve the efficiency of translation termination.

[0149] The nucleic acid encoding the consensus Tyr antigen can also encode an immunoglobulin E (IgE) leader sequence. The nucleic acid encoding the consensus Tyr antigen can further encode the 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 by a peptide bond. The nucleic acid encoding the consensus Tyr antigen can also include the nucleotide sequence encoding the IgE leader sequence. In some embodiments, the nucleic acid encoding the consensus Tyr antigen does not have, i.e., does not include, the nucleotide sequence encoding the IgE leader sequence.

[0150] The consensus Tyr antigen can be nucleic acid sequence SEQ ID NO:1, and SEQ ID NO:1 encodes amino acid sequence SEQ ID NO:2. SEQ ID NO:1 encodes a consensus Tyr protein that is linked to the IgE leader sequence. The consensus Tyr protein can be linked to the IgE leader sequence and an HA tag. In other embodiments, the consensus Tyr protein may not have the IgE leader sequence and / or the HA tag, i.e., may not be linked to the sequence and / or the tag.

[0151] In some embodiments, the consensus Tyr antigen can be a nucleic acid sequence having 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% identity over the full length of the nucleic acid sequence shown in SEQ ID NO: 1. In other embodiments, the consensus Tyr antigen can be a nucleic acid sequence encoding an amino acid sequence having 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% identity over the full length of the amino acid sequence shown in SEQ ID NO: 2. The consensus Tyr antigen can be an amino acid sequence having 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% identity over the full length of the amino acid sequence shown in SEQ ID NO: 2.

[0152] Some embodiments relate to proteins homologous to the Tyr consensus protein, immunogenic fragments of the Tyr consensus protein, and nucleic acid sequences encoding 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 of the proteins shown herein and immunogenic fragments of proteins homologous to the proteins shown herein are also provided.

[0153] Some embodiments relate to nucleic acid molecules encoding immunogenic proteins having 95% homology to the nucleic acid coding sequences herein. Some embodiments relate to nucleic acid molecules encoding immunogenic proteins having 96% homology to the nucleic acid coding sequences herein. Some embodiments relate to nucleic acid molecules encoding immunogenic proteins having 97% homology to the nucleic acid coding sequences herein. Some embodiments relate to nucleic acid molecules encoding immunogenic proteins having 98% homology to the nucleic acid coding sequences herein. Some embodiments relate to nucleic acid molecules encoding immunogenic proteins having 99% homology to the nucleic acid coding sequences herein. In some embodiments, a nucleic acid molecule having a coding sequence disclosed herein that is homologous to the coding sequence of the consensus protein disclosed herein includes a sequence encoding an IgE leader sequence that is linked to the 5' end of the coding sequence encoding the homologous protein sequence disclosed herein.

[0154] Some embodiments relate to nucleic acid sequences encoding proteins having a specified identity, expressed as a percentage relative to the full-length Tyr consensus protein, immunogenic fragments of the Tyr consensus protein, and immunogenic fragments of proteins having identity to the Tyr consensus protein. Such nucleic acid molecules can encode immunogenic proteins having up to 80% identity to the full-length Tyr consensus sequence, up to 85% identity to the full-length 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. Similarly, nucleic acid sequences encoding immunogenic fragments of proteins having identity, expressed as a percentage similar to that shown above, to the immunogenic fragments shown herein and to the Tyr proteins shown herein are also provided.

[0155] In some embodiments, the nucleic acid sequence does not have a coding sequence encoding a leader sequence. In some embodiments, the nucleic acid sequence does not have a coding sequence encoding an IgE leader.

[0156] Some embodiments relate to the fragment of SEQ ID NO: 1. 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: 1. The fragment can be at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% homologous to the fragment of SEQ ID NO: 1. 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 the fragment of SEQ ID NO: 1. In some embodiments, the fragment comprises a sequence encoding a leader sequence that is an immunoglobulin leader such as, for example, an IgE leader. In some embodiments, the fragment does not have a coding sequence encoding a leader sequence. In some embodiments, the fragment does not have a coding sequence encoding a leader sequence such as, for example, an IgE leader.

[0157] Furthermore, 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 is SEQ ID NO: 2. The amino acid sequence of the consensus Tyr protein linked to the IgE leader may be linked to an HA tag.

[0158] Some embodiments relate to a protein homologous to SEQ ID NO: 2. Some embodiments relate to an immunogenic protein having 95% homology to the consensus protein sequence set forth in SEQ ID NO: 2. Some embodiments relate to an immunogenic protein having 96% homology to the consensus protein sequence set forth in SEQ ID NO: 2. Some embodiments relate to an immunogenic protein having 97% homology to the consensus protein sequence set forth in SEQ ID NO: 2. Some embodiments relate to an immunogenic protein having 98% homology to the consensus protein sequence set forth in SEQ ID NO: 2. Some embodiments relate to an immunogenic protein having 99% homology to the consensus protein sequence set forth in SEQ ID NO: 2.

[0159] Some embodiments relate to a protein that is identical to SEQ ID NO: 2. Some embodiments relate to an immunogenic protein having an amino acid sequence that is 80% identical to the full-length consensus amino acid sequence set forth in SEQ ID NO: 2. Some embodiments relate to an immunogenic protein having an amino acid sequence that is 85% identical to the full-length consensus amino acid sequence set forth in SEQ ID NO: 2. Some embodiments relate to an immunogenic protein having an amino acid sequence that is 90% identical to the full-length consensus amino acid sequence set forth in SEQ ID NO: 2. Some embodiments relate to an immunogenic protein having an amino acid sequence that is 91% identical to the full-length consensus amino acid sequence set forth in SEQ ID NO: 2. Some embodiments relate to an immunogenic protein having an amino acid sequence that is 92% identical to the full-length consensus amino acid sequence set forth in SEQ ID NO: 2. Some embodiments relate to an immunogenic protein having an amino acid sequence that is 93% identical to the full-length consensus amino acid sequence set forth in SEQ ID NO: 2. Some embodiments relate to an immunogenic protein having an amino acid sequence that is 94% identical to the full-length consensus amino acid sequence set forth in SEQ ID NO: 2. Some embodiments relate to an immunogenic protein having an amino acid sequence that is 95% identical to the full-length consensus amino acid sequence set forth in SEQ ID NO: 2. Some embodiments relate to an immunogenic protein having an amino acid sequence that is 96% identical to the full-length consensus amino acid sequence set forth in SEQ ID NO: 2. Some embodiments relate to an immunogenic protein having an amino acid sequence that is 97% identical to the full-length consensus amino acid sequence set forth in SEQ ID NO: 2. Some embodiments relate to an immunogenic protein having an amino acid sequence that is 98% identical to the full-length consensus amino acid sequence set forth in SEQ ID NO: 2. Some embodiments relate to an immunogenic protein having an amino acid sequence that is 99% identical to the full-length consensus amino acid sequence set forth in SEQ ID NO: 2.

[0160] In some embodiments, the protein does not have a leader sequence. In some embodiments, the protein does not have an IgE leader. Fragments 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: 2 can be provided. The immunogenic fragment 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 SEQ ID NO: 2. In some embodiments, the fragment comprises a leader sequence that is an immunoglobulin leader such as, for example, an IgE leader. In some embodiments, the fragment does not have a leader sequence. In some embodiments, the fragment does not have a leader sequence such as, for example, an IgE leader.

[0161] An immunogenic fragment of a protein having an amino acid sequence homologous to the immunogenic fragment of SEQ ID NO: 2 can be provided. Such an immunogenic fragment 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 that is at least 95% homologous to SEQ ID NO: 2. Some embodiments relate to an immunogenic fragment having 96% homology to an immunogenic fragment of a consensus protein sequence herein. Some embodiments relate to an immunogenic fragment having 97% homology to an immunogenic fragment of a consensus protein sequence herein. Some embodiments relate to an immunogenic fragment having 98% homology to an immunogenic fragment of a consensus protein sequence herein. Some embodiments relate to an immunogenic fragment having 99% homology to an immunogenic fragment of a consensus protein sequence herein. In some embodiments, the fragment comprises a leader sequence that is an immunoglobulin leader, such as an IgE leader. In some embodiments, the fragment does not have a leader sequence. In some embodiments, the fragment does not have a leader sequence such as an IgE leader, for example.

[0162] It is possible to provide an immunogenic fragment of a protein having the same amino acid sequence as the immunogenic fragment of SEQ ID NO: 2. Such an immunogenic fragment is 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% identical to the amino acid sequence shown in SEQ ID NO: 2. In some embodiments, the fragment comprises a leader sequence that is an immunoglobulin leader, such as an IgE leader. In some embodiments, the fragment does not have a leader sequence. In some embodiments, the fragment does not have a leader sequence such as an IgE leader.

[0163] As referred to herein with respect to the ligation of a signal peptide or leader sequence to the N-terminus of a protein, the signal peptide / leader sequence replaces the N-terminal methionine of the protein encoded by the nucleic acid sequence encoding a protein that does not have a signal peptide coding sequence, by the start codon.

[0164] (2) Tyrosinase-related protein 1 (TYRP1) The immunogenic composition of the present invention can include the cancer antigen tyrosinase-related protein 1 (TYRP1), a fragment thereof, or a variant thereof. TYRP1 is encoded by the TYRP1 gene and is a 75 kDa transmembrane glycoprotein that is expressed in both normal and malignant melanogenic cells and melanoma cells. TYRP1, like tyrosinase, contains a modified called the M box that can bind to the microphthalmia transcription factor (MITF), and MITF plays a central role in melanogenic cells in pigmentation, cell proliferation, and differentiation. TYRP1 can help stabilize tyrosinase and can form heterodimers, and the dimers can prevent early death of melanogenic cells by attenuating tyrosinase-mediated cytotoxicity.

[0165] As described above for tyrosinase, tyrosinase-related protein 1 (TYRP-1) is also involved in the synthesis and pigment mechanism of melanin in melanogenic cells and can be recognized by the immune system in subjects affected by melanoma. Thus, TYRP-1 can be an antigen related to melanoma.

[0166] The TYRP-1 antigen can induce antigen-specific T cells and / or a high-titer antibody response, thereby inducing or eliciting an immune response that targets or is reactive to cancer or tumors expressing said 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-gamma (IFN-γ) and / or tumor necrosis factor alpha (TNF-α). In other embodiments, the induced or elicited immune response can include, but is not limited to, factors that downregulate presentation by MHC, antigen-specific regulatory T cells (Tregs), cytokines such as PD-L1, FasL, IL-10, and TGF-β, tumor-associated macrophages, tumor-associated fibroblasts, soluble factors produced by immunosuppressive cells, CTLA-4, PD-1, MDSC, MCP-1, and factors that upregulate immune checkpoint molecules, etc. One or more immunosuppressive factors that promote the growth of tumors or cancers expressing said antigen can be reduced or inhibited, and the immune checkpoint molecules will be described in more detail later.

[0167] The TYRP-1 antigen can include protein epitopes that make the antigen particularly effective as an immunogen capable of inducing an anti-TYRP-1 immune response against the antigen. The TYRP-1 antigen can include the full-length translation product, variants thereof, fragments thereof, or combinations thereof. The TYRP-1 antigen can include a consensus protein.

[0168] The nucleic acid sequence encoding the consensus TYRP-1 antigen can be optimized with respect to codon usage frequency and the corresponding RNA transcript. The nucleic acid encoding the consensus TYRP-1 antigen can be codons and RNA optimized for expression. In some embodiments, the nucleic acid sequence encoding the consensus TYRP-1 antigen can include a Kozak sequence (e.g., GCC ACC) to improve translation efficiency. The nucleic acid encoding the consensus TYRP-1 antigen can include multiple stop codons (e.g., TGA TGA) to improve the efficiency of translation termination.

[0169] The nucleic acid encoding the consensus TYRP-1 antigen can also encode an immunoglobulin E (IgE) leader sequence. The nucleic acid encoding the consensus TYRP-1 antigen can 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 can 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, i.e., does not include, a nucleotide sequence encoding the IgE leader sequence.

[0170] The consensus TYRP-1 antigen can be nucleic acid sequence SEQ ID NO: 3, and SEQ ID NO: 3 encodes amino acid sequence SEQ ID NO: 4. SEQ ID NO: 3 encodes a 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 have, i.e., may not be linked to, an IgE leader sequence and / or an HA tag.

[0171] In some embodiments, the consensus TYRP-1 antigen can be a nucleic acid sequence having 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% identity over the full 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 encoding an amino acid sequence having 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% identity over the full length of the amino acid sequence shown in SEQ ID NO: 4. The consensus TYRP-1 antigen can be an amino acid sequence having 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% identity over the full length of the amino acid sequence shown in SEQ ID NO: 4.

[0172] Some embodiments relate to proteins homologous to the TYRP-1 consensus protein, immunogenic fragments of the TYRP-1 consensus protein, and nucleic acid sequences encoding 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 of the proteins shown herein and immunogenic fragments of proteins homologous to the proteins shown herein are also provided.

[0173] Some embodiments relate to nucleic acid molecules encoding immunogenic proteins having 95% homology to the nucleic acid coding sequences herein. Some embodiments relate to nucleic acid molecules encoding immunogenic proteins having 96% homology to the nucleic acid coding sequences herein. Some embodiments relate to nucleic acid molecules encoding immunogenic proteins having 97% homology to the nucleic acid coding sequences herein. Some embodiments relate to nucleic acid molecules encoding immunogenic proteins having 98% homology to the nucleic acid coding sequences herein. Some embodiments relate to nucleic acid molecules encoding immunogenic proteins having 99% homology to the nucleic acid coding sequences herein. In some embodiments, a nucleic acid molecule having a coding sequence disclosed herein that is homologous to the coding sequence of the consensus protein disclosed herein includes a sequence encoding an IgE leader sequence that is linked to the 5' end of the coding sequence encoding the homologous protein sequence disclosed herein.

[0174] Some embodiments relate to proteins having a specified identity expressed as a percentage to the full-length TYRP-1 consensus protein, immunogenic fragments of the TYRP-1 consensus protein, and nucleic acid sequences encoding immunogenic fragments of proteins having identity to the TYRP-1 consensus protein. Such nucleic acid molecules can 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. Similarly, nucleic acid sequences encoding immunogenic fragments of proteins having an identity expressed as a percentage similar to that shown above with respect to the immunogenic fragments shown herein and the TYRP-1 protein shown herein are also provided.

[0175] In some embodiments, the nucleic acid sequence does not have a coding sequence encoding a leader sequence. In some embodiments, the nucleic acid sequence does not have a coding sequence encoding an IgE leader.

[0176] Some embodiments relate to the fragment of SEQ ID NO: 3. 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: 3. The fragment can be at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% homologous to the fragment of SEQ ID NO: 3. 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 the fragment of SEQ ID NO: 3. In some embodiments, the fragment comprises a sequence encoding a leader sequence, which is an immunoglobulin leader such as, for example, an IgE leader. In some embodiments, the fragment does not have a coding sequence encoding a leader sequence. In some embodiments, the fragment does not have a coding sequence encoding a leader sequence such as, for example, an IgE leader.

[0177] Furthermore, 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 is SEQ ID NO: 4. The amino acid sequence of the consensus TYRP-1 protein linked to the IgE leader may be linked to the HA tag.

[0178] Some embodiments relate to a protein homologous to SEQ ID NO: 4. Some embodiments relate to an immunogenic protein having 95% homology to the consensus protein sequence set forth in SEQ ID NO: 4. Some embodiments relate to an immunogenic protein having 96% homology to the consensus protein sequence set forth in SEQ ID NO: 4. Some embodiments relate to an immunogenic protein having 97% homology to the consensus protein sequence set forth in SEQ ID NO: 4. Some embodiments relate to an immunogenic protein having 98% homology to the consensus protein sequence set forth in SEQ ID NO: 4. Some embodiments relate to an immunogenic protein having 99% homology to the consensus protein sequence set forth in SEQ ID NO: 4.

[0179] Some embodiments relate to a protein that is 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 set forth 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 set forth 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 set forth 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 set forth 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 set forth 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 set forth in SEQ ID NO: 4. Some embodiments relate to an immunogenic protein having an amino acid sequence that is 94% identical to the full-length consensus amino acid sequence set forth in SEQ ID NO: 4. Some embodiments relate to an immunogenic protein having an amino acid sequence that is 95% identical to the full-length consensus amino acid sequence set forth in SEQ ID NO: 4. Some embodiments relate to an immunogenic protein having an amino acid sequence that is 96% identical to the full-length consensus amino acid sequence set forth in SEQ ID NO: 4. Some embodiments relate to an immunogenic protein having an amino acid sequence that is 97% identical to the full-length consensus amino acid sequence set forth in SEQ ID NO: 4. Some embodiments relate to an immunogenic protein having an amino acid sequence that is 98% identical to the full-length consensus amino acid sequence set forth in SEQ ID NO: 4. Some embodiments relate to an immunogenic protein having an amino acid sequence that is 99% identical to the full-length consensus amino acid sequence set forth in SEQ ID NO: 4.

[0180] In some embodiments, the protein does not have a leader sequence. In some embodiments, the protein does not have an IgE leader. Fragments 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 TYRP-1 protein. An immunogenic fragment of SEQ ID NO: 4 can be provided. The immunogenic fragment 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 SEQ ID NO: 4. In some embodiments, the fragment comprises a leader sequence that is an immunoglobulin leader, such as, for example, an IgE leader. In some embodiments, the fragment does not have a leader sequence. In some embodiments, the fragment does not have a leader sequence such as, for example, an IgE leader.

[0181] It is possible to provide an immunogenic fragment of a protein having an amino acid sequence homologous to the immunogenic fragment of SEQ ID NO: 4. Such an immunogenic fragment 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 that is at least 95% homologous to SEQ ID NO: 4. Some embodiments relate to an immunogenic fragment having 96% homology to the immunogenic fragment of the consensus protein sequence herein. Some embodiments relate to an immunogenic fragment having 97% homology to the immunogenic fragment of the consensus protein sequence herein. Some embodiments relate to an immunogenic fragment having 98% homology to the immunogenic fragment of the consensus protein sequence herein. Some embodiments relate to an immunogenic fragment having 99% homology to the immunogenic fragment of the consensus protein sequence herein. In some embodiments, the fragment comprises a leader sequence that is an immunoglobulin leader, such as an IgE leader. In some embodiments, the fragment does not have a leader sequence. In some embodiments, the fragment does not have a leader sequence such as an IgE leader.

[0182] It is possible to provide an immunogenic fragment of a protein having the same amino acid sequence as the immunogenic fragment of SEQ ID NO: 4. Such an immunogenic fragment may 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% identical to the amino acid sequence shown in SEQ ID NO: 4, of a protein. In some embodiments, the fragment comprises a leader sequence that is an immunoglobulin leader such as, for example, an IgE leader. In some embodiments, the fragment does not have a leader sequence. In some embodiments, the fragment does not have a leader sequence such as, for example, an IgE leader.

[0183] As referred to herein with respect to the ligation of a signal peptide or leader sequence to the N-terminus of a protein, the signal peptide / leader sequence replaces the N-terminal methionine of the protein encoded by the start codon of the nucleic acid sequence encoding a protein that does not have a signal peptide coding sequence.

[0184] (3) Tyrosinase-related protein 2 (TYRP2) The immunogenic composition of the present invention can comprise the cancer antigen tyrosinase-related protein 2 (TYRP2, also known as dopachrome tautomerase (DCT)), a fragment thereof, or a variant thereof. TYRP2 / DCT is a protein encoded by the TYRP2 / DCT gene and composed of 519 amino acids, and is expressed in both normal and malignant melanogenic cells and melanoma cells. TYRP2 / DCT is a well-characterized melanogenic cell-specific enzyme that functions, in cooperation with tyrosinase and TYRP1, in the conversion of L-tyrosine to melanin in melanogenic cells. 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 described above) to form eumelanin. Studies have shown that TYRP2 / DCT can be a mediator of drug resistance in melanoma cells, having specificity for DNA-damaging agents. Since TYRP2 / DCT has often been reported to be highly expressed in melanoma, this melanogenic cell-specific enzyme plays an important role in contributing to the intrinsic drug resistance phenotype of melanoma to various anti-cancer DNA-damaging agents.

[0185] As described above for tyrosinase, tyrosinase-related protein 2 (TYRP-2) can also be involved in the synthesis of melanin and can be recognized by the immune system in subjects affected by melanoma. Furthermore, TYRP-2 can mediate drug resistance in melanoma cells. Thus, TYRP-2 can be an antigen associated with melanoma.

[0186] The TYRP-2 antigen can induce an antigen-specific T cell and / or high-titer antibody response, thereby inducing or eliciting an immune response that targets or is reactive to a cancer or tumor expressing the above 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-gamma (IFN-γ) and / or tumor necrosis factor alpha (TNF-α). In other embodiments, the induced or elicited immune response includes, but is not limited to, factors that downregulate presentation by MHC, antigen-specific regulatory T cells (Tregs), cytokines such as PD-L1, FasL, IL-10, and TGF-β, tumor-associated macrophages, tumor-associated fibroblasts, soluble factors produced by immunosuppressive cells, factors that upregulate CTLA-4, PD-1, MDSC, MCP-1, and immune checkpoint molecules, etc. One or more immunosuppressive factors that promote the growth of tumors or cancers expressing the above antigen can be reduced or inhibited, and the immune checkpoint molecules will be described in more detail later.

[0187] The TYRP-2 antigen can contain a protein epitope that makes the antigen particularly effective as an immunogen capable of inducing an anti-TYRP-2 immune response against the antigen. The TYRP-2 antigen can include the full-length translation product, variants thereof, fragments thereof, or combinations thereof. The TYRP-2 antigen can include a consensus protein.

[0188] The nucleic acid sequence encoding the consensus TYRP-2 antigen can be optimized with respect to codon usage frequency and the corresponding RNA transcript. The nucleic acid encoding the consensus TYRP-2 antigen can be codons and RNA optimized for expression. In some embodiments, the nucleic acid sequence encoding the consensus TYRP-2 antigen can include a Kozak sequence (e.g., GCC ACC) to improve translation efficiency. The nucleic acid encoding the consensus TYRP-2 antigen can include multiple stop codons (e.g., TGA TGA) to improve the efficiency of translation termination.

[0189] The nucleic acid encoding the consensus TYRP-2 antigen can also encode an immunoglobulin E (IgE) leader sequence. The nucleic acid encoding the consensus TYRP-2 antigen can further encode the 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-2 antigen by a peptide bond. The nucleic acid encoding the consensus TYRP-2 antigen can also include the nucleotide sequence encoding the IgE leader sequence. In some embodiments, the nucleic acid encoding the consensus TYRP-2 antigen does not have, i.e., does not include, the nucleotide sequence encoding the IgE leader sequence.

[0190] The consensus TYRP-2 antigen can be nucleic acid sequence SEQ ID NO: 5, and SEQ ID NO: 5 encodes the amino acid sequence SEQ ID NO: 6. SEQ ID NO: 5 encodes a consensus TYRP-2 protein linked to an IgE leader sequence. The consensus TYRP-2 protein can be linked to an IgE leader sequence and an HA tag. In other embodiments, the consensus TYRP-2 protein may not have, i.e., may not be linked to, an IgE leader sequence and / or an HA tag.

[0191] In some embodiments, the consensus TYRP-2 antigen can be a nucleic acid sequence having 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% identity over the full length of the nucleic acid sequence shown in SEQ ID NO: 5. In other embodiments, the consensus TYRP-2 antigen can be a nucleic acid sequence encoding an amino acid sequence having 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% identity over the full length of the amino acid sequence shown in SEQ ID NO: 6. The consensus TYRP-2 antigen can be an amino acid sequence having 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% identity over the full length of the amino acid sequence shown in SEQ ID NO: 6.

[0192] Some embodiments relate to proteins homologous to the TYRP-2 consensus protein, immunogenic fragments of the TYRP-2 consensus protein, and nucleic acid sequences encoding 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 of the proteins shown herein and immunogenic fragments of proteins homologous to the proteins shown herein are also provided.

[0193] Some embodiments relate to nucleic acid molecules encoding immunogenic proteins having 95% homology to the nucleic acid coding sequences herein. Some embodiments relate to nucleic acid molecules encoding immunogenic proteins having 96% homology to the nucleic acid coding sequences herein. Some embodiments relate to nucleic acid molecules encoding immunogenic proteins having 97% homology to the nucleic acid coding sequences herein. Some embodiments relate to nucleic acid molecules encoding immunogenic proteins having 98% homology to the nucleic acid coding sequences herein. Some embodiments relate to nucleic acid molecules encoding immunogenic proteins having 99% homology to the nucleic acid coding sequences herein. In some embodiments, a nucleic acid molecule having a coding sequence disclosed herein that is homologous to the coding sequence of the consensus protein disclosed herein includes a sequence encoding an IgE leader sequence that is linked to the 5' end of the coding sequence encoding the homologous protein sequence disclosed herein.

[0194] Some embodiments relate to a protein having a specific identity expressed as a percentage relative to the full-length TYRP-2 consensus protein, an immunogenic fragment of the TYRP-2 consensus protein, and a nucleic acid sequence encoding an immunogenic fragment of a protein having identity to the TYRP-2 consensus protein. Such nucleic acid molecules can encode immunogenic proteins having up to 80% identity to the full-length TYRP-2 consensus sequence, up to 85% identity to the full-length TYRP-2 consensus sequence, up to 90% identity to the full-length TYRP-2 consensus sequence, up to 91% identity to the full-length TYRP-2 consensus sequence, up to 92% identity to the full-length TYRP-2 consensus sequence, up to 93% identity to the full-length TYRP-2 consensus sequence, up to 94% identity to the full-length TYRP-2 consensus sequence, up to 95% identity to the full-length TYRP-2 consensus sequence, up to 96% identity to the full-length TYRP-2 consensus sequence, up to 97% identity to the full-length TYRP-2 consensus sequence, up to 98% identity to the full-length TYRP-2 consensus sequence, and up to 99% identity to the full-length TYRP-2 consensus sequence. Similarly, nucleic acid sequences encoding immunogenic fragments of proteins having an identity expressed as a percentage similar to that shown above with respect to the immunogenic fragments shown herein and the TYRP-2 proteins shown herein are also provided.

[0195] In some embodiments, the nucleic acid sequence does not have a coding sequence encoding a leader sequence. In some embodiments, the nucleic acid sequence does not have a coding sequence encoding an IgE leader.

[0196] Some embodiments relate to the fragment of SEQ ID NO: 5. 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: 5. The fragment can be at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% homologous to the fragment of SEQ ID NO: 5. 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 the fragment of SEQ ID NO: 5. In some embodiments, the fragment comprises a sequence encoding a leader sequence that is an immunoglobulin leader such as, for example, an IgE leader. In some embodiments, the fragment does not have a coding sequence encoding a leader sequence. In some embodiments, the fragment does not have a coding sequence encoding a leader sequence such as, for example, an IgE leader.

[0197] Furthermore, the amino acid sequence of the consensus TYRP-2 protein is SEQ ID NO: 6. The amino acid sequence of the consensus TYRP-2 protein linked to an IgE leader is SEQ ID NO: 6. The amino acid sequence of the consensus TYRP-2 protein linked to an IgE leader may be linked to an HA tag.

[0198] Some embodiments relate to a protein homologous to SEQ ID NO: 2. Some embodiments relate to an immunogenic protein having 95% homology to the consensus protein sequence set forth in SEQ ID NO: 6. Some embodiments relate to an immunogenic protein having 96% homology to the consensus protein sequence set forth in SEQ ID NO: 6. Some embodiments relate to an immunogenic protein having 97% homology to the consensus protein sequence set forth in SEQ ID NO: 6. Some embodiments relate to an immunogenic protein having 98% homology to the consensus protein sequence set forth in SEQ ID NO: 6. Some embodiments relate to an immunogenic protein having 99% homology to the consensus protein sequence set forth in SEQ ID NO: 6.

[0199] Some embodiments relate to a protein that is identical to SEQ ID NO: 6. Some embodiments relate to an immunogenic protein having an amino acid sequence that is 80% identical to the full-length consensus amino acid sequence set forth in SEQ ID NO: 6. Some embodiments relate to an immunogenic protein having an amino acid sequence that is 85% identical to the full-length consensus amino acid sequence set forth in SEQ ID NO: 6. Some embodiments relate to an immunogenic protein having an amino acid sequence that is 90% identical to the full-length consensus amino acid sequence set forth in SEQ ID NO: 6. Some embodiments relate to an immunogenic protein having an amino acid sequence that is 91% identical to the full-length consensus amino acid sequence set forth in SEQ ID NO: 6. Some embodiments relate to an immunogenic protein having an amino acid sequence that is 92% identical to the full-length consensus amino acid sequence set forth in SEQ ID NO: 6. Some embodiments relate to an immunogenic protein having an amino acid sequence that is 93% identical to the full-length consensus amino acid sequence set forth in SEQ ID NO: 6. Some embodiments relate to an immunogenic protein having an amino acid sequence that is 94% identical to the full-length consensus amino acid sequence set forth in SEQ ID NO: 6. Some embodiments relate to an immunogenic protein having an amino acid sequence that is 95% identical to the full-length consensus amino acid sequence set forth in SEQ ID NO: 6. Some embodiments relate to an immunogenic protein having an amino acid sequence that is 96% identical to the full-length consensus amino acid sequence set forth in SEQ ID NO: 6. Some embodiments relate to an immunogenic protein having an amino acid sequence that is 97% identical to the full-length consensus amino acid sequence set forth in SEQ ID NO: 6. Some embodiments relate to an immunogenic protein having an amino acid sequence that is 98% identical to the full-length consensus amino acid sequence set forth in SEQ ID NO: 6. Some embodiments relate to an immunogenic protein having an amino acid sequence that is 99% identical to the full-length consensus amino acid sequence set forth in SEQ ID NO: 6.

[0200] In some embodiments, the protein does not have a leader sequence. In some embodiments, the protein does not have an IgE leader. Fragments 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 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 SEQ ID NO: 6. In some embodiments, the fragment comprises a leader sequence that is an immunoglobulin leader, such as, for example, an IgE leader. In some embodiments, the fragment does not have a leader sequence. In some embodiments, the fragment does not have a leader sequence such as, for example, an IgE leader.

[0201] It is possible to provide an immunogenic fragment of a protein having an amino acid sequence homologous to the immunogenic fragment of SEQ ID NO: 6. Such an immunogenic fragment 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 that is 95% or more homologous to SEQ ID NO: 6. Some embodiments relate to an immunogenic fragment having 96% homology to an immunogenic fragment of a consensus protein sequence herein. Some embodiments relate to an immunogenic fragment having 97% homology to an immunogenic fragment of a consensus protein sequence herein. Some embodiments relate to an immunogenic fragment having 98% homology to an immunogenic fragment of a consensus protein sequence herein. Some embodiments relate to an immunogenic fragment having 99% homology to an immunogenic fragment of a consensus protein sequence herein. In some embodiments, the fragment comprises a leader sequence that is an immunoglobulin leader, such as an IgE leader. In some embodiments, the fragment does not have a leader sequence. In some embodiments, the fragment does not have a leader sequence such as an IgE leader, for example.

[0202] It is possible to provide an immunogenic fragment of a protein having the same amino acid sequence as the immunogenic fragment of SEQ ID NO: 6. Such an immunogenic fragment is 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% identical to the amino acid sequence shown in SEQ ID NO: 6. In some embodiments, the fragment comprises a leader sequence that is an immunoglobulin leader such as, for example, an IgE leader. In some embodiments, the fragment does not have a leader sequence. In some embodiments, the fragment does not have a leader sequence such as, for example, an IgE leader.

[0203] As referred to herein with respect to the ligation of a signal peptide or leader sequence to the N-terminus of a protein, the signal peptide / leader sequence replaces the N-terminal methionine of the protein encoded by the start codon of the nucleic acid sequence encoding a protein that does not have a signal peptide coding sequence.

[0204] (4) Melanoma-associated Antigen 4 (MAGEA4) The immunogenic composition of the present invention can contain a cancer antigen melanoma-associated antigen 4 (MAGEA4), a fragment thereof, or a variant thereof. MAGE-A4 is a protein encoded by the MAGE-A4 gene and composed of 317 amino acids, and is expressed in male germ cells and tumor cells of various histological types such as digestive tract cancer, esophageal cancer, and lung cancer. MAGE-A4 binds to a tumor protein, ganp. This specific binding to MAGE-A4 is mediated by its C-terminus. Studies have shown that exogenous MAGE-A4 can partially inhibit the adhesion-independent growth of ganp-overexpressing cells in vitro and can inhibit the formation of tumors metastasized from these cells in nude mice. This inhibition depends on the binding between MAGE-A4 and ganp, suggesting that the interaction between ganp and MAGE-A4 inhibits ganp-mediated carcinogenesis. The expression of MAGE in tumor tissues is a result rather than a cause of tumorigenesis, and the MAGE gene appears to be involved in the immune process by targeting and destroying early tumor cells.

[0205] Melanoma-associated antigen 4 (MAGEA4) may be involved in embryonic development and 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 various tumors.

[0206] The MAGEA4 antigen can induce antigen-specific T cells and / or high-titer antibody responses, thereby inducing or eliciting an immune response that targets or is reactive to cancer or tumors expressing said 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-gamma (IFN-γ) and / or tumor necrosis factor alpha (TNF-α). In other embodiments, the induced or elicited immune response can include, but is not limited to, factors that downregulate presentation by MHC, antigen-specific regulatory T cells (Tregs), cytokines such as PD-L1, FasL, IL-10, and TGF-β, tumor-associated macrophages, tumor-associated fibroblasts, soluble factors produced by immunosuppressive cells, factors that upregulate immune checkpoint molecules such as CTLA-4, PD-1, MDSC, MCP-1, and one or more immunosuppressive factors that promote the growth of tumors or cancers expressing said antigen can be reduced or inhibited, and the immune checkpoint molecules will be described in more detail later.

[0207] The MAGEA4 antigen can include protein epitopes that make the antigen particularly effective as an immunogen capable of inducing an anti-MAGEA4 immune response against the antigen. The MAGEA4 antigen can include the full-length translation product, variants thereof, fragments thereof, or combinations thereof. The MAGEA4 antigen can include a consensus protein.

[0208] The nucleic acid sequence encoding the consensus MAGEA4 antigen can be optimized with respect to codon usage frequency and the corresponding RNA transcript. The nucleic acid encoding the consensus MAGEA4 antigen can be codons 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 improve translation efficiency. The nucleic acid encoding the consensus MAGEA4 antigen can include multiple stop codons (e.g., TGA TGA) to improve the efficiency of translation termination.

[0209] The nucleic acid encoding the consensus MAGEA4 antigen can also encode an immunoglobulin E (IgE) leader sequence. The nucleic acid encoding the consensus Tyr antigen can 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 by a peptide bond. The nucleic acid encoding the consensus MAGEA4 antigen can also include a nucleotide sequence encoding an IgE leader sequence. In some embodiments, the nucleic acid encoding the consensus MAGEA4 antigen does not have, i.e., does not include, a nucleotide sequence encoding an IgE leader sequence.

[0210] The consensus MAGEA4 antigen can be nucleic acid sequence SEQ ID NO: 7, and SEQ ID NO: 7 encodes the amino acid sequence SEQ ID NO: 8. SEQ ID NO: 7 encodes a 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 have an IgE leader sequence and / or an HA tag, i.e., may not be linked to the sequence and / or the tag.

[0211] In some embodiments, the consensus MAGEA4 antigen can be a nucleic acid sequence having 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% identity over the full length of the nucleic acid sequence shown in SEQ ID NO: 7. In other embodiments, the consensus MAGEA4 antigen can be a nucleic acid sequence encoding an amino acid sequence having 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% identity over the full length of the amino acid sequence shown in SEQ ID NO: 8. The consensus MAGEA4 antigen can be an amino acid sequence having 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% identity over the full length of the amino acid sequence shown in SEQ ID NO: 8.

[0212] Some embodiments relate to proteins homologous to the MAGEA4 consensus protein, immunogenic fragments of the MAGEA4 consensus protein, and nucleic acid sequences encoding 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 of the proteins shown herein and immunogenic fragments of proteins homologous to the proteins shown herein are also provided.

[0213] Some embodiments relate to nucleic acid molecules encoding immunogenic proteins having 95% homology to the nucleic acid coding sequences herein. Some embodiments relate to nucleic acid molecules encoding immunogenic proteins having 96% homology to the nucleic acid coding sequences herein. Some embodiments relate to nucleic acid molecules encoding immunogenic proteins having 97% homology to the nucleic acid coding sequences herein. Some embodiments relate to nucleic acid molecules encoding immunogenic proteins having 98% homology to the nucleic acid coding sequences herein. Some embodiments relate to nucleic acid molecules encoding immunogenic proteins having 99% homology to the nucleic acid coding sequences herein. In some embodiments, a nucleic acid molecule having a coding sequence disclosed herein that is homologous to the coding sequence of the consensus protein disclosed herein includes a sequence encoding an IgE leader sequence that is linked to the 5' end of the coding sequence encoding the homologous protein sequence disclosed herein.

[0214] Some embodiments relate to nucleic acid sequences encoding proteins having a specified identity expressed as a percentage 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 encode immunogenic proteins having up to 80% identity to the full-length MAGEA4 consensus sequence, up to 85% identity to the full-length MAGEA4 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. Similarly, nucleic acid sequences encoding immunogenic fragments of proteins having identity expressed as a percentage similar to that shown above to the immunogenic fragments shown herein and the MAGEA4 protein shown herein are also provided.

[0215] In some embodiments, the nucleic acid sequence does not have a coding sequence encoding a leader sequence. In some embodiments, the nucleic acid sequence does not have a coding sequence encoding an IgE leader.

[0216] Some embodiments relate to the fragment of SEQ ID NO: 7. 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: 7. The fragment can be at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% homologous to the fragment of SEQ ID NO: 7. 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 the fragment of SEQ ID NO: 7. In some embodiments, the fragment comprises a sequence encoding a leader sequence, such as an immunoglobulin leader such as an IgE leader. In some embodiments, the fragment does not have a coding sequence encoding a leader sequence. In some embodiments, the fragment does not have a coding sequence encoding a leader sequence, such as an IgE leader.

[0217] Furthermore, 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 is SEQ ID NO: 8. The amino acid sequence of the consensus MAGEA4 protein linked to the IgE leader may be linked to an HA tag.

[0218] Some embodiments relate to a protein homologous to SEQ ID NO: 8. Some embodiments relate to an immunogenic protein having 95% homology to the consensus protein sequence set forth in SEQ ID NO: 8. Some embodiments relate to an immunogenic protein having 96% homology to the consensus protein sequence set forth in SEQ ID NO: 8. Some embodiments relate to an immunogenic protein having 97% homology to the consensus protein sequence set forth in SEQ ID NO: 8. Some embodiments relate to an immunogenic protein having 98% homology to the consensus protein sequence set forth in SEQ ID NO: 8. Some embodiments relate to an immunogenic protein having 99% homology to the consensus protein sequence set forth in SEQ ID NO: 8.

[0219] Some embodiments relate to a protein that is identical to SEQ ID NO: 8. Some embodiments relate to an immunogenic protein having an amino acid sequence that is 80% identical to the full-length consensus amino acid sequence set forth in SEQ ID NO: 8. Some embodiments relate to an immunogenic protein having an amino acid sequence that is 85% identical to the full-length consensus amino acid sequence set forth in SEQ ID NO: 8. Some embodiments relate to an immunogenic protein having an amino acid sequence that is 90% identical to the full-length consensus amino acid sequence set forth in SEQ ID NO: 8. Some embodiments relate to an immunogenic protein having an amino acid sequence that is 91% identical to the full-length consensus amino acid sequence set forth in SEQ ID NO: 8. Some embodiments relate to an immunogenic protein having an amino acid sequence that is 92% identical to the full-length consensus amino acid sequence set forth in SEQ ID NO: 8. Some embodiments relate to an immunogenic protein having an amino acid sequence that is 93% identical to the full-length consensus amino acid sequence set forth in SEQ ID NO: 8. Some embodiments relate to an immunogenic protein having an amino acid sequence that is 94% identical to the full-length consensus amino acid sequence set forth in SEQ ID NO: 8. Some embodiments relate to an immunogenic protein having an amino acid sequence that is 95% identical to the full-length consensus amino acid sequence set forth in SEQ ID NO: 8. Some embodiments relate to an immunogenic protein having an amino acid sequence that is 96% identical to the full-length consensus amino acid sequence set forth in SEQ ID NO: 8. Some embodiments relate to an immunogenic protein having an amino acid sequence that is 97% identical to the full-length consensus amino acid sequence set forth in SEQ ID NO: 8. Some embodiments relate to an immunogenic protein having an amino acid sequence that is 98% identical to the full-length consensus amino acid sequence set forth in SEQ ID NO: 8. Some embodiments relate to an immunogenic protein having an amino acid sequence that is 99% identical to the full-length consensus amino acid sequence set forth in SEQ ID NO: 8.

[0220] In some embodiments, the protein does not have a leader sequence. In some embodiments, the protein does not have an IgE leader. Fragments 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 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 SEQ ID NO: 8. In some embodiments, the fragment comprises a leader sequence that is an immunoglobulin leader, such as, for example, an IgE leader. In some embodiments, the fragment does not have a leader sequence. In some embodiments, the fragment does not have a leader sequence such as, for example, an IgE leader.

[0221] It is possible to provide an immunogenic fragment of a protein having an amino acid sequence homologous to the immunogenic fragment of SEQ ID NO: 8. Such an immunogenic fragment 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 that is at least 95% homologous to SEQ ID NO: 8. Some embodiments relate to an immunogenic fragment having 96% homology to the immunogenic fragment of the consensus protein sequence herein. Some embodiments relate to an immunogenic fragment having 97% homology to the immunogenic fragment of the consensus protein sequence herein. Some embodiments relate to an immunogenic fragment having 98% homology to the immunogenic fragment of the consensus protein sequence herein. Some embodiments relate to an immunogenic fragment having 99% homology to the immunogenic fragment of the consensus protein sequence herein. In some embodiments, the fragment comprises a leader sequence that is an immunoglobulin leader, such as an IgE leader. In some embodiments, the fragment does not have a leader sequence. In some embodiments, the fragment does not have a leader sequence such as an IgE leader, for example.

[0222] An immunogenic fragment of a protein having the same amino acid sequence as the immunogenic fragment of SEQ ID NO: 8 can be provided. Such an immunogenic fragment 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 that is 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence shown in SEQ ID NO: 8. In some embodiments, the fragment comprises a leader sequence that is an immunoglobulin leader such as, for example, an IgE leader. In some embodiments, the fragment does not have a leader sequence. In some embodiments, the fragment does not have a leader sequence such as, for example, an IgE leader.

[0223] As referred to herein with respect to the ligation of a signal peptide or leader sequence to the N-terminus of a protein, the signal peptide / leader sequence replaces the N-terminal methionine of the protein encoded by the nucleic acid sequence encoding a protein that does not have a signal peptide coding sequence by the start codon.

[0224] (5) Growth hormone releasing hormone (GHRH) The immunogenic composition of the present invention can comprise a cancer antigen growth hormone-releasing hormone (GHRH, also known as growth-hormone-releasing factor (GRF or GHRF) or somatocrinin), 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 from the pituitary gland, growth, metabolism, and the release of regulators of body structure. GHRH also stimulates the production of growth hormone. Antagonists of GHRH inhibit the growth of various cancers such as, for example, osteosarcoma, glioblastoma, prostate cancer, renal cancer, pancreatic cancer, colorectal cancer, and breast cancer. Thus, GHRH can be an antigen associated with various tumors.

[0225] The GHRH antigen can induce an antigen-specific T cell and / or high-titer antibody response, thereby inducing or eliciting an immune response directed against or reactive to a cancer or tumor expressing said 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-gamma (IFN-γ) and / or tumor necrosis factor alpha (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 a tumor or cancer expressing said antigen, such as, but not limited to, factors that downregulate presentation by MHC, antigen-specific regulatory T cells (Tregs), cytokines such as PD-L1, FasL, IL-10, and TGF-β, tumor-associated macrophages, tumor-associated fibroblasts, soluble factors produced by immunosuppressive cells, factors that upregulate CTLA-4, PD-1, MDSC, MCP-1, and immune checkpoint molecules, which are described in more detail later.

[0226] The GHRH antigen can include protein epitopes that make the antigen particularly effective as an immunogen capable of inducing an anti-GHRH immune response that counteracts the antigen. The GHRH antigen can include the full-length translation product, variants thereof, fragments thereof, or combinations thereof. The GHRH antigen can include a consensus protein.

[0227] The nucleic acid sequence encoding the consensus GHRH antigen can be optimized with respect to codon usage frequency and the corresponding RNA transcript. 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 improve the efficiency of translation. The nucleic acid encoding the consensus GHRH antigen can include multiple stop codons (e.g., TGA TGA) to improve the efficiency of translation termination.

[0228] The nucleic acid encoding the consensus GHRH antigen can also encode an immunoglobulin E (IgE) leader sequence. The nucleic acid encoding the consensus GHRH antigen can further encode the 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 by a peptide bond. The nucleic acid encoding the consensus GHRH antigen can also include the nucleotide sequence encoding the IgE leader sequence. In some embodiments, the nucleic acid encoding the consensus GHRH antigen does not have, i.e., does not include, the nucleotide sequence encoding the IgE leader sequence.

[0229] The consensus GHRH antigen can have the nucleic acid sequence SEQ ID NO: 9, and SEQ ID NO: 9 encodes the amino acid sequence SEQ ID NO: 10. SEQ ID NO: 9 encodes a 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 have an IgE leader sequence and / or an HA tag, that is, it may not be linked to the sequence and / or the tag.

[0230] In some embodiments, the consensus GHRH antigen can be a nucleic acid sequence having 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% identity over the full 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 encoding an amino acid sequence having 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% identity over the full length of the amino acid sequence shown in SEQ ID NO: 10. The consensus GHRH antigen can be an amino acid sequence having 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% identity over the full length of the amino acid sequence shown in SEQ ID NO: 10.

[0231] Some embodiments relate to proteins homologous to the GHRH consensus protein, immunogenic fragments of the GHRH consensus protein, and nucleic acid sequences encoding immunogenic fragments of homologous proteins. Such nucleic acid molecules can 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 the immunogenic fragments shown herein and immunogenic fragments of proteins homologous to the proteins shown herein are also provided.

[0232] Some embodiments relate to nucleic acid molecules encoding immunogenic proteins having 95% homology to the nucleic acid coding sequences herein. Some embodiments relate to nucleic acid molecules encoding immunogenic proteins having 96% homology to the nucleic acid coding sequences herein. Some embodiments relate to nucleic acid molecules encoding immunogenic proteins having 97% homology to the nucleic acid coding sequences herein. Some embodiments relate to nucleic acid molecules encoding immunogenic proteins having 98% homology to the nucleic acid coding sequences herein. Some embodiments relate to nucleic acid molecules encoding immunogenic proteins having 99% homology to the nucleic acid coding sequences herein. In some embodiments, a nucleic acid molecule having the coding sequence disclosed herein, homologous to the coding sequence of the consensus protein disclosed herein, includes a sequence encoding an IgE leader sequence that is linked to the 5' end of the coding sequence encoding the homologous protein sequence disclosed herein.

[0233] Some embodiments relate to a protein having a specific identity expressed as a percentage to a full-length GHRH consensus protein, an immunogenic fragment of the GHRH consensus protein, and a nucleic acid sequence encoding an immunogenic fragment of a protein having identity to the GHRH consensus protein. Such nucleic acid molecules can encode immunogenic proteins having up to 80% identity to the full-length GHRH consensus sequence, up to 85% identity to the full-length GHRH 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 of proteins having identity expressed as a percentage similar to that shown above to the immunogenic fragments shown herein and to the GHRH proteins shown herein are also provided.

[0234] In some embodiments, the nucleic acid sequence does not have a coding sequence encoding a leader sequence. In some embodiments, the nucleic acid sequence does not have a coding sequence encoding an IgE leader.

[0235] Some embodiments relate to the fragment of SEQ ID NO: 9. 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: 9. The fragment can be at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% homologous to the fragment of SEQ ID NO: 9. 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 the fragment of SEQ ID NO: 9. In some embodiments, the fragment comprises a sequence encoding a leader sequence that is an immunoglobulin leader, such as an IgE leader. In some embodiments, the fragment does not have a coding sequence encoding a leader sequence. In some embodiments, the fragment does not have a coding sequence encoding a leader sequence, such as an IgE leader.

[0236] Furthermore, 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 is SEQ ID NO: 10. The amino acid sequence of the consensus GHRH protein linked to the IgE leader may be linked to an HA tag.

[0237] Some embodiments relate to a protein homologous to SEQ ID NO: 10. Some embodiments relate to an immunogenic protein having 95% homology to the consensus protein sequence set forth in SEQ ID NO: 10. Some embodiments relate to an immunogenic protein having 96% homology to the consensus protein sequence set forth in SEQ ID NO: 10. Some embodiments relate to an immunogenic protein having 97% homology to the consensus protein sequence set forth in SEQ ID NO: 10. Some embodiments relate to an immunogenic protein having 98% homology to the consensus protein sequence set forth in SEQ ID NO: 10. Some embodiments relate to an immunogenic protein having 99% homology to the consensus protein sequence set forth in SEQ ID NO: 10.

[0238] Some embodiments relate to a protein identical to SEQ ID NO: 10. Some embodiments relate to an immunogenic protein having an amino acid sequence that is 80% identical to the full-length consensus amino acid sequence set forth in SEQ ID NO: 10. Some embodiments relate to an immunogenic protein having an amino acid sequence that is 85% identical to the full-length consensus amino acid sequence set forth in SEQ ID NO: 10. Some embodiments relate to an immunogenic protein having an amino acid sequence that is 90% identical to the full-length consensus amino acid sequence set forth in SEQ ID NO: 10. Some embodiments relate to an immunogenic protein having an amino acid sequence that is 91% identical to the full-length consensus amino acid sequence set forth in SEQ ID NO: 10. Some embodiments relate to an immunogenic protein having an amino acid sequence that is 92% identical to the full-length consensus amino acid sequence set forth in SEQ ID NO: 10. Some embodiments relate to an immunogenic protein having an amino acid sequence that is 93% identical to the full-length consensus amino acid sequence set forth in SEQ ID NO: 10. Some embodiments relate to an immunogenic protein having an amino acid sequence that is 94% identical to the full-length consensus amino acid sequence set forth in SEQ ID NO: 10. Some embodiments relate to an immunogenic protein having an amino acid sequence that is 95% identical to the full-length consensus amino acid sequence set forth in SEQ ID NO: 10. Some embodiments relate to an immunogenic protein having an amino acid sequence that is 96% identical to the full-length consensus amino acid sequence set forth in SEQ ID NO: 10. Some embodiments relate to an immunogenic protein having an amino acid sequence that is 97% identical to the full-length consensus amino acid sequence set forth in SEQ ID NO: 10. Some embodiments relate to an immunogenic protein having an amino acid sequence that is 98% identical to the full-length consensus amino acid sequence set forth in SEQ ID NO: 10. Some embodiments relate to an immunogenic protein having an amino acid sequence that is 99% identical to the full-length consensus amino acid sequence set forth in SEQ ID NO: 10.

[0239] In some embodiments, the protein does not have a leader sequence. In some embodiments, the protein does not have an IgE leader. Fragments 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 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 SEQ ID NO: 10. In some embodiments, the fragment comprises a leader sequence that is an immunoglobulin leader, such as, for example, an IgE leader. In some embodiments, the fragment does not have a leader sequence. In some embodiments, the fragment does not have a leader sequence such as, for example, an IgE leader.

[0240] An immunogenic fragment of a protein having an amino acid sequence homologous to the immunogenic fragment of SEQ ID NO: 10 can be provided. Such an immunogenic fragment 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 that is at least 95% homologous to SEQ ID NO: 10. Some embodiments relate to an immunogenic fragment having 96% homology to an immunogenic fragment of a consensus protein sequence herein. Some embodiments relate to an immunogenic fragment having 97% homology to an immunogenic fragment of a consensus protein sequence herein. Some embodiments relate to an immunogenic fragment having 98% homology to an immunogenic fragment of a consensus protein sequence herein. Some embodiments relate to an immunogenic fragment having 99% homology to an immunogenic fragment of a consensus protein sequence herein. In some embodiments, the fragment comprises a leader sequence that is an immunoglobulin leader, such as an IgE leader. In some embodiments, the fragment does not have a leader sequence. In some embodiments, the fragment does not have a leader sequence such as an IgE leader, for example.

[0241] An immunogenic fragment of a protein having the same amino acid sequence as the immunogenic fragment of SEQ ID NO: 10 can be provided. Such an immunogenic fragment 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 that is 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence set forth in SEQ ID NO: 10. In some embodiments, the fragment comprises a leader sequence that is an immunoglobulin leader, such as an IgE leader. In some embodiments, the fragment does not have a leader sequence. In some embodiments, the fragment does not have a leader sequence such as an IgE leader.

[0242] As referred to herein with respect to the ligation of a signal peptide or leader sequence to the N-terminus of a protein, the signal peptide / leader sequence replaces the N-terminal methionine of the protein encoded by the start codon of the nucleic acid sequence encoding a protein that does not have a signal peptide coding sequence.

[0243] (6) MART-1 / Melan-A The immunogenic composition of the present invention can include the cancer antigen MART-1 (also known as Melan-A), its fragments, or its variants. MART-1 is encoded by the MLANA gene, is a protein of 118 amino acids, contains a single transmembrane domain, and is expressed in most melanoma cells. MART-1 forms a complex with structural proteins and affects its expression, stability, transport, and the processing necessary for the structure and maturation of melanosomes. Therefore, MART-1 is essential for regulating mammalian pigmentation. Defects in melanosome maturation have been associated with susceptibility to cancer. MART-1 can be expressed in a number of cancers, including but not limited to melanoma.

[0244] Melan-A, also known as melanoma antigen recognized by T cells (MART-1), is a differentiation antigen of melanocytes and can be present in normal skin, retina, and melanocytes. Melan-A can be associated with the endoplasmic reticulum and melanosomes. Melan-A can be recognized by cytotoxic T cells as an antigen on melanoma cells, but can also be associated with other tumors of melanocyte origin or with their differentiation, such as clear cell sarcoma and melanotic schwannoma (i.e., cells having melanosomes). Therefore, Melan-A can be an antigen associated with various tumors derived from cells having melanosomes.

[0245] The Melan-A antigen can induce antigen-specific T cell and / or high-titer antibody responses, thereby inducing or eliciting an immune response that targets or is reactive to cancer or tumors expressing the above 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-gamma (IFN-γ) and / or tumor necrosis factor alpha (TNF-α). In other embodiments, the induced or elicited immune response can include, but is not limited to, factors that downregulate presentation by MHC, antigen-specific regulatory T cells (Tregs), cytokines such as PD-L1, FasL, IL-10, and TGF-β, tumor-associated macrophages, tumor-associated fibroblasts, soluble factors produced by immunosuppressive cells, and factors that upregulate one or more immunosuppressive factors that promote the growth of tumors or cancers expressing the above antigen, such as CTLA-4, PD-1, MDSC, MCP-1, and factors that upregulate immune checkpoint molecules, which will be described in more detail later.

[0246] The Melan-A antigen can contain protein epitopes that make the antigen particularly effective as an immunogen capable of inducing an anti-Melan-A immune response against the antigen. The Melan-A antigen can include the full-length translation product, variants thereof, fragments thereof, or combinations thereof. The Melan-A antigen can include a consensus protein.

[0247] The nucleic acid sequence encoding the consensus MELAN-A antigen can be optimized with respect to codon usage and the corresponding RNA transcript. 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 improve translation efficiency. The nucleic acid encoding the consensus MELAN-A antigen can include multiple stop codons (e.g., TGA TGA) to improve the efficiency of translation termination.

[0248] The nucleic acid encoding the consensus MELAN-A antigen can also encode an immunoglobulin E (IgE) leader sequence. The nucleic acid encoding the consensus MELAN-A antigen can 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 by a peptide bond. The nucleic acid encoding the consensus MELAN-A antigen can 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, i.e., does not include, a nucleotide sequence encoding the IgE leader sequence.

[0249] The consensus MELAN-A antigen can be nucleic acid sequence SEQ ID NO: 11, and SEQ ID NO: 11 encodes amino acid sequence SEQ ID NO: 12. SEQ ID NO: 11 encodes a 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 have, i.e., may not be linked to, an IgE leader sequence and / or an HA tag.

[0250] In some embodiments, the consensus Melan-A antigen can be a nucleic acid sequence having 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% identity over the full 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 encoding an amino acid sequence having 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% identity over the full length of the amino acid sequence shown in SEQ ID NO: 12. The consensus Melan-A antigen can be an amino acid sequence having 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% identity over the full length of the amino acid sequence shown in SEQ ID NO: 12.

[0251] Some embodiments relate to proteins homologous to the Melan-A consensus protein, immunogenic fragments of the Melan-A consensus protein, and nucleic acid sequences encoding 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 of the proteins shown herein and immunogenic fragments of proteins homologous to the proteins shown herein are also provided.

[0252] Some embodiments relate to nucleic acid molecules encoding immunogenic proteins having 95% homology to the nucleic acid coding sequences herein. Some embodiments relate to nucleic acid molecules encoding immunogenic proteins having 96% homology to the nucleic acid coding sequences herein. Some embodiments relate to nucleic acid molecules encoding immunogenic proteins having 97% homology to the nucleic acid coding sequences herein. Some embodiments relate to nucleic acid molecules encoding immunogenic proteins having 98% homology to the nucleic acid coding sequences herein. Some embodiments relate to nucleic acid molecules encoding immunogenic proteins having 99% homology to the nucleic acid coding sequences herein. In some embodiments, a nucleic acid molecule having a coding sequence disclosed herein that is homologous to the coding sequence of the consensus protein disclosed herein includes a sequence encoding an IgE leader sequence that is 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 identity, expressed as a percentage relative to the full-length Melan-A consensus protein, immunogenic fragments of the Melan-A consensus protein, and immunogenic fragments of proteins having identity to the Melan-A consensus protein. Such nucleic acid molecules can 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. Similarly, nucleic acid sequences encoding immunogenic fragments of proteins having identity, expressed as a percentage similar to that shown above, to the immunogenic fragments shown herein and to the Melan-A protein shown herein are also provided.

[0254] In some embodiments, the nucleic acid sequence does not have a coding sequence encoding a leader sequence. In some embodiments, the nucleic acid sequence does not have a coding sequence encoding an IgE leader.

[0255] Some embodiments relate to the fragment of SEQ ID NO: 11. 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: 11. The fragment can be at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% homologous to the fragment of SEQ ID NO: 11. 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 the fragment of SEQ ID NO: 11. In some embodiments, the fragment comprises a coding sequence encoding a leader sequence that is an immunoglobulin leader, such as an IgE leader. In some embodiments, the fragment does not have a coding sequence encoding a leader sequence. In some embodiments, the fragment does not have a coding sequence encoding a leader sequence, such as an IgE leader.

[0256] Furthermore, 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 is SEQ ID NO: 12. The amino acid sequence of the consensus melan-A protein linked to the IgE leader may be linked to an HA tag.

[0257] Some embodiments relate to a protein homologous to SEQ ID NO: 12. Some embodiments relate to an immunogenic protein having 95% homology to the consensus protein sequence set forth in SEQ ID NO: 12. Some embodiments relate to an immunogenic protein having 96% homology to the consensus protein sequence set forth in SEQ ID NO: 12. Some embodiments relate to an immunogenic protein having 97% homology to the consensus protein sequence set forth in SEQ ID NO: 12. Some embodiments relate to an immunogenic protein having 98% homology to the consensus protein sequence set forth in SEQ ID NO: 12. Some embodiments relate to an immunogenic protein having 99% homology to the consensus protein sequence set forth in SEQ ID NO: 12.

[0258] Some embodiments relate to a protein that is 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 set forth 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 set forth 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 set forth 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 set forth 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 set forth 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 set forth in SEQ ID NO: 12. Some embodiments relate to an immunogenic protein having an amino acid sequence that is 94% identical to the full-length consensus amino acid sequence set forth in SEQ ID NO: 12. Some embodiments relate to an immunogenic protein having an amino acid sequence that is 95% identical to the full-length consensus amino acid sequence set forth in SEQ ID NO: 12. Some embodiments relate to an immunogenic protein having an amino acid sequence that is 96% identical to the full-length consensus amino acid sequence set forth in SEQ ID NO: 12. Some embodiments relate to an immunogenic protein having an amino acid sequence that is 97% identical to the full-length consensus amino acid sequence set forth in SEQ ID NO: 12. Some embodiments relate to an immunogenic protein having an amino acid sequence that is 98% identical to the full-length consensus amino acid sequence set forth in SEQ ID NO: 12. Some embodiments relate to an immunogenic protein having an amino acid sequence that is 99% identical to the full-length consensus amino acid sequence set forth in SEQ ID NO: 12.

[0259] In some embodiments, the protein does not have a leader sequence. In some embodiments, the protein does not have an IgE leader. Fragments 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 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 SEQ ID NO: 12. In some embodiments, the fragment comprises a leader sequence that is an immunoglobulin leader, such as, for example, an IgE leader. In some embodiments, the fragment does not have a leader sequence. In some embodiments, the fragment does not have a leader sequence such as, for example, an IgE leader.

[0260] It is possible to provide an immunogenic fragment of a protein having an amino acid sequence homologous to the immunogenic fragment of SEQ ID NO: 12. Such an immunogenic fragment 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 that is at least 95% homologous to SEQ ID NO: 12. Some embodiments relate to an immunogenic fragment having 96% homology to the immunogenic fragment of the consensus protein sequence herein. Some embodiments relate to an immunogenic fragment having 97% homology to the immunogenic fragment of the consensus protein sequence herein. Some embodiments relate to an immunogenic fragment having 98% homology to the immunogenic fragment of the consensus protein sequence herein. Some embodiments relate to an immunogenic fragment having 99% homology to the immunogenic fragment of the consensus protein sequence herein. In some embodiments, the fragment comprises a leader sequence that is an immunoglobulin leader, such as an IgE leader. In some embodiments, the fragment does not have a leader sequence. In some embodiments, the fragment does not have a leader sequence such as an IgE leader.

[0261] An immunogenic fragment of a protein having the same amino acid sequence as the immunogenic fragment of SEQ ID NO: 12 can be provided. Such an immunogenic fragment 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 that is 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence shown in SEQ ID NO: 12. In some embodiments, the fragment comprises a leader sequence that is an immunoglobulin leader, such as an IgE leader. In some embodiments, the fragment does not have a leader sequence. In some embodiments, the fragment does not have a leader sequence such as an IgE leader.

[0262] As referred to herein with respect to the ligation of a signal peptide or leader sequence to the N-terminus of a protein, the signal peptide / leader sequence replaces the N-terminal methionine of the protein encoded by the start codon of a nucleic acid sequence encoding a protein that does not have a signal peptide coding sequence.

[0263] (7)NY-ESO-1 The immunogenic composition of the present invention can contain cancer antigen New York esophageal cancer-1 (also called NY-ESO-1, CTAG1), its fragment, or its variant. NY-ESO-1 is encoded by the CTAG1B gene and is a long-chain protein of 180 amino acids with a glycine-rich N-terminal region and an extremely hydrophobic C-terminal region. NY-ESO-1 is expressed in a restricted manner in normal tissues but often occurs in cancers. NY-ESO-1 can be expressed in a number of cancers including, but not limited to, bladder cancer, colorectal cancer, esophageal cancer, gastric cancer, hepatocellular carcinoma, head and neck cancer, melanoma, non-small cell lung cancer, ovarian cancer, pancreatic cancer, synovial cancer, and prostate cancer.

[0264] Cancer / testis antigen (NY-ESO-1) can be expressed in the testis and ovary. NY-ESO-1 is associated with various cancers and can induce a humoral immune response. A subject suffering from cancer or a tumor can become immunogenic to NY-ESO-1. Thus, NY-ESO-1 can be an antigen associated with various tumors.

[0265] The NY-ESO-1 antigen can induce antigen-specific T cell and / or high-titer antibody responses, thereby inducing or eliciting an immune response directed against or reactive to a cancer or tumor expressing said 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-gamma (IFN-γ) and / or tumor necrosis factor alpha (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 a tumor or cancer expressing said antigen, including, but not limited to, factors that downregulate presentation by MHC, antigen-specific regulatory T cells (Tregs), cytokines such as PD-L1, FasL, IL-10, and TGF-β, tumor-associated macrophages, tumor-associated fibroblasts, soluble factors produced by immunosuppressive cells, factors that upregulate CTLA-4, PD-1, MDSC, MCP-1, and immune checkpoint molecules, which are described in more detail below.

[0266] The NY-ESO-1 antigen can increase the cellular immune response in a subject administered with the NY-ESO-1 antigen 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, as compared to the cellular immune response in a subject not administered with the NY-ESO-1 antigen. In some embodiments, the NY-ESO-1 antigen can increase the cellular immune response 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, 4500-fold, 4600-fold, 4700-fold, 4800-fold, 4900-fold, 5000-fold, 5100-fold, 5200-fold, 5300-fold, 5400-fold, 5500-fold, 5600-fold, 5700-fold, 5800-fold, 5900-fold, or 6000-fold, as compared to the cellular immune response in a subject not administered with the NY-ESO-1 antigen.

[0267] The NY-ESO-1 antigen can increase the concentration of interferon-gamma (IFN-γ) in a subject administered with the NY-ESO-1 antigen 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, as compared to the IFN-γ concentration in a subject not administered with the NY-ESO-1 antigen. In some embodiments, the NY-ESO-1 antigen can increase the IFN-γ concentration 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, 4500-fold, 4600-fold, 4700-fold, 4800-fold, 4900-fold, 5000-fold, 5100-fold, 5200-fold, 5300-fold, 5400-fold, 5500-fold, 5600-fold, 5700-fold, 5800-fold, 5900-fold, or 6000-fold, as compared to the IFN-γ concentration in a subject not administered with the NY-ESO-1 antigen.

[0268] The NY-ESO-1 antigen can contain a protein epitope that makes the antigen particularly effective as an immunogen capable of inducing an anti-NY-ESO-1 immune response that counteracts the antigen. The NY-ESO-1 antigen can include the full-length translation product, variants thereof, fragments thereof, or combinations thereof. The NY-ESO-1 antigen can include a consensus protein.

[0269] The nucleic acid sequence encoding the consensus NY-ESO-1 antigen can be optimized with respect to codon usage frequency and the corresponding RNA transcript. The nucleic acid encoding the consensus NY-ESO-1 antigen can be codons 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 improve translation efficiency. The nucleic acid encoding the consensus NY-ESO-1 antigen can include multiple stop codons (e.g., TGA TGA) to improve the efficiency of translation termination.

[0270] The nucleic acid encoding the consensus NY-ESO-1 antigen can also encode an immunoglobulin E (IgE) leader sequence. The nucleic acid encoding the consensus NY-ESO-1 antigen can 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 by a peptide bond. The nucleic acid encoding the consensus NY-ESO-1 antigen can also include a nucleotide sequence encoding the IgE leader sequence. In some embodiments, the nucleic acid encoding the consensus NY-ESO-1 antigen does not have, i.e., does not include, a nucleotide sequence encoding the IgE leader sequence.

[0271] The consensus NY-ESO-1 antigen can be nucleic acid sequence SEQ ID NO: 13, and SEQ ID NO: 13 encodes the amino acid sequence SEQ ID NO: 14. SEQ ID NO: 13 encodes a 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 have an IgE leader sequence and / or an HA tag, i.e., may not be linked to the sequence and / or the tag.

[0272] In some embodiments, the consensus NY-ESO-1 antigen can be a nucleic acid sequence having 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% identity over the full 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 encoding an amino acid sequence having 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% identity over the full length of the amino acid sequence shown in SEQ ID NO: 14. The consensus NY-ESO-1 antigen can be an amino acid sequence having 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% identity over the full length of the amino acid sequence shown in SEQ ID NO: 14.

[0273] Some embodiments relate to proteins homologous to the NY-ESO-1 consensus protein, immunogenic fragments of the NY-ESO-1 consensus protein, and nucleic acid sequences encoding 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 of the proteins shown herein and immunogenic fragments of proteins homologous to the proteins shown herein are also provided.

[0274] Some embodiments relate to nucleic acid molecules encoding immunogenic proteins having 95% homology to the nucleic acid coding sequences herein. Some embodiments relate to nucleic acid molecules encoding immunogenic proteins having 96% homology to the nucleic acid coding sequences herein. Some embodiments relate to nucleic acid molecules encoding immunogenic proteins having 97% homology to the nucleic acid coding sequences herein. Some embodiments relate to nucleic acid molecules encoding immunogenic proteins having 98% homology to the nucleic acid coding sequences herein. Some embodiments relate to nucleic acid molecules encoding immunogenic proteins having 99% homology to the nucleic acid coding sequences herein. In some embodiments, a nucleic acid molecule having a coding sequence disclosed herein that is homologous to the coding sequence of the consensus protein disclosed herein includes a sequence encoding an IgE leader sequence that is linked to the 5' end of the coding sequence encoding the homologous protein sequence disclosed herein.

[0275] Some embodiments relate to nucleic acid sequences encoding proteins having a specified identity expressed as a percentage relative to the full-length NY-ESO-1 consensus protein, immunogenic fragments of the NY-ESO-1 consensus protein, and immunogenic fragments of proteins having identity to the NY-ESO-1 consensus protein. Such nucleic acid molecules can 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 NY-ESO-1 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. Similarly, nucleic acid sequences encoding immunogenic fragments of proteins having identity expressed as a percentage similar to that shown above with respect to the immunogenic fragments shown herein and the NY-ESO-1 protein shown herein are also provided.

[0276] In some embodiments, the nucleic acid sequence does not have a coding sequence encoding a leader sequence. In some embodiments, the nucleic acid sequence does not have a coding sequence encoding an IgE leader.

[0277] Some embodiments relate to the fragment of SEQ ID NO: 13. 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: 13. The fragment can be at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% homologous to the fragment of SEQ ID NO: 13. 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 the fragment of SEQ ID NO: 13. In some embodiments, the fragment comprises a sequence encoding a leader sequence, which is an immunoglobulin leader such as, for example, an IgE leader. In some embodiments, the fragment does not have a coding sequence encoding a leader sequence. In some embodiments, the fragment does not have a coding sequence encoding a leader sequence such as, for example, an IgE leader.

[0278] Furthermore, 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 an IgE leader is SEQ ID NO: 14. The amino acid sequence of the consensus NY-ESO-1 protein linked to an IgE leader may be linked to an HA tag.

[0279] Some embodiments relate to a protein homologous to SEQ ID NO: 14. Some embodiments relate to an immunogenic protein having 95% homology to the consensus protein sequence set forth in SEQ ID NO: 14. Some embodiments relate to an immunogenic protein having 96% homology to the consensus protein sequence set forth in SEQ ID NO: 14. Some embodiments relate to an immunogenic protein having 97% homology to the consensus protein sequence set forth in SEQ ID NO: 14. Some embodiments relate to an immunogenic protein having 98% homology to the consensus protein sequence set forth in SEQ ID NO: 14. Some embodiments relate to an immunogenic protein having 99% homology to the consensus protein sequence set forth in SEQ ID NO: 14.

[0280] Some embodiments relate to a protein that is 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 set forth 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 set forth 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 set forth 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 set forth 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 set forth 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 set forth in SEQ ID NO: 14. Some embodiments relate to an immunogenic protein having an amino acid sequence that is 94% identical to the full-length consensus amino acid sequence set forth in SEQ ID NO: 14. Some embodiments relate to an immunogenic protein having an amino acid sequence that is 95% identical to the full-length consensus amino acid sequence set forth in SEQ ID NO: 14. Some embodiments relate to an immunogenic protein having an amino acid sequence that is 96% identical to the full-length consensus amino acid sequence set forth in SEQ ID NO: 14. Some embodiments relate to an immunogenic protein having an amino acid sequence that is 97% identical to the full-length consensus amino acid sequence set forth in SEQ ID NO: 14. Some embodiments relate to an immunogenic protein having an amino acid sequence that is 98% identical to the full-length consensus amino acid sequence set forth in SEQ ID NO: 14. Some embodiments relate to an immunogenic protein having an amino acid sequence that is 99% identical to the full-length consensus amino acid sequence set forth in SEQ ID NO: 14.

[0281] In some embodiments, the protein does not have a leader sequence. In some embodiments, the protein does not have an IgE leader. Fragments 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 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 SEQ ID NO: 14. In some embodiments, the fragment comprises a leader sequence that is an immunoglobulin leader, such as, for example, an IgE leader. In some embodiments, the fragment does not have a leader sequence. In some embodiments, the fragment does not have a leader sequence such as, for example, an IgE leader.

[0282] An immunogenic fragment of a protein having an amino acid sequence homologous to the immunogenic fragment of SEQ ID NO: 14 can be provided. Such an immunogenic fragment 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 that is at least 95% homologous to SEQ ID NO: 14. Some embodiments relate to an immunogenic fragment having 96% homology to an immunogenic fragment of a consensus protein sequence herein. Some embodiments relate to an immunogenic fragment having 97% homology to an immunogenic fragment of a consensus protein sequence herein. Some embodiments relate to an immunogenic fragment having 98% homology to an immunogenic fragment of a consensus protein sequence herein. Some embodiments relate to an immunogenic fragment having 99% homology to an immunogenic fragment of a consensus protein sequence herein. In some ...

Claims

1. An immunogenic composition comprising a nucleic acid molecule encoding a consensus TERT antigen, wherein the consensus TERT antigen is 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 at least 95% identity with 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; 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, the fragment comprising at least 95% of the full-length amino acid sequence; and the immunogenic composition comprising an amino acid sequence selected from the group consisting of

2. 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 at least 95% identity with 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; 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, the fragment comprising at least 95% of the full-length amino acid sequence; and further comprising one or more nucleotide sequences encoding one or more amino acid sequences selected from the group consisting of the immunogenic composition according to claim 1.

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

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

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

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

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

9. The nucleic acid molecule is 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 at least 95% identity with 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; 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. The immunogenic composition according to claim 1, comprising a nucleotide sequence selected from the group consisting of the foregoing.

10. 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 at least 95% identity with 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; 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, said fragment comprising at least 95% of the full-length nucleotide sequence; The immunogenic composition according to claim 2, comprising one or more nucleotide sequences selected from the group consisting of the above.

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

12. The immunogenic composition according to claim 1, comprising one or more plasmids.

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

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

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

16. The method according to claim 15, wherein the administration comprises an electroporation step.

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

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

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

20. The method according to 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) 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 at least 95% identity with 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; 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, said fragment comprising at least 95% of the full-length nucleotide sequence; a nucleic acid molecule comprising one or more nucleotide sequences selected from the group consisting of.

22. 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 at least 95% identity with 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; 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, said fragment comprising at least 95% of the full-length nucleotide sequence; the nucleic acid molecule according to claim 21, further comprising one or more nucleotide sequences selected from the group consisting of.

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

24. 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 more identity with 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; 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, the fragment containing at least 95% of the full-length amino acid sequence, an amino acid molecule comprising one or more amino acid sequences selected from the group consisting of. **Claim 25** 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 more identity with 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; 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, the fragment containing at least 95% of the full-length amino acid sequence, the amino acid molecule according to claim 24, further comprising one or more amino acid sequences selected from the group consisting of.

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