Breast and ovarian cancer vaccines

Abstract

To provide a composition comprising an epitope of a peptide that may elicit an immune response in a subject following administration.SOLUTION: Such a composition may comprise a nucleic acid. The composition may comprise a peptide. The method disclosed herein comprises a step of administering a composition comprising an epitope of a peptide to a subject in need thereof. In some of aspects, the composition disclosed herein comprises [a first plasmid comprising] a first nucleotide sequence encoding a first epitope of a first antigen expressed by a cell associated with breast cancer, and a second plasmid comprising a second nucleotide sequence encoding a second epitope of a second antigen expressed by a cell associated with breast cancer, where the first and second nucleotide sequences are located in one or more plasmids.SELECTED DRAWING: None

Description

Technical Field

[0001] Cross-reference This application claims the benefit of U.S. Provisional Patent Application No. 61 / 972,176, filed on Mar. 28, 2014, which is hereby incorporated by reference in its entirety.

[0002] Statement Regarding Federally Sponsored Research This invention was made with government support under Award Numbers W81XWH-11-1-0760 from the Department of Defense, P50CA083636 from the National Cancer Institute, and R01CA098761 from the National Cancer Institute.

Background Art

[0003] Cancer therapies have conventionally been achieved by surgical reduction of tumor mass and subsequent chemotherapy and / or radiation therapy. This strategy can reduce tumors and, in still non-progressive stages, often results in complete remission. Unfortunately, the prognosis for more advanced tumors has hardly changed over the past 50 years, and a significant proportion of cancer-related deaths are caused by subsequent metastases. New preventive and therapeutic treatments are needed to combat the increasing incidence of cancer.

[0004] Worldwide, more than a million people are diagnosed with breast cancer each year, and more than 400,000 people die of breast cancer each year. It is estimated that 1 in 8 women will be diagnosed with breast cancer at some point in their lifetime. Prevention of breast cancer onset could have significant health and economic benefits for every individual. Billions of dollars could be saved if there were no need to undergo expensive cancer-related screening and treatment interventions. New approaches for breast cancer prevention and treatment are needed.

Summary of the Invention

Means for Solving the Problems

[0005] In some embodiments, the composition described herein includes a first plasmid comprising a first nucleotide sequence encoding a first epitope of a first antigen expressed by cells associated with breast cancer, and a second nucleotide sequence encoding a second epitope of a second antigen expressed by cells associated with breast cancer, wherein the first nucleotide sequence and the second nucleotide sequence are located on one or more plasmids. The present invention provides, for example, the following items. (Item 1) a) An isolated and purified plasmid comprising a nucleotide sequence encoding a polypeptide, wherein the polypeptide comprises a plurality of epitopes; and b) An excipient. A composition comprising the same. (Item 2) The composition according to Item 1, wherein the plurality of epitopes comprise one or more epitopes having at least 90% sequence identity to an amino acid sequence selected from SEQ ID NOs: 1, 6, 8-10, 14-16, 20, 25-28, 32-34, 46-56, 60-62, 66-75, 82-85, or 87. (Item 3) The composition according to Item 1 or 2, wherein the plurality of epitopes comprise one or more epitopes having at least 90% sequence identity to an amino acid sequence selected from SEQ ID NOs: 82-84. (Item 4) The composition according to Item 1 or 2, wherein the plurality of epitopes comprise one or more epitopes having at least 90% sequence identity to an amino acid sequence selected from SEQ ID NOs: 1, 6, 8-10, 14-16, 20, 25-28, or 32-34. (Item 5) The composition according to Item 1 or 2, wherein the plurality of epitopes comprise one or more epitopes having at least 90% sequence identity to an amino acid sequence selected from SEQ ID NOs: 46-56, 60-62, or 66-75. (Item 6) The composition according to item 1 or 2, wherein the plurality of epitopes comprises one or more epitopes having at least 90% sequence identity to an amino acid sequence selected from SEQ ID NO: 54, 73, 85 or 87. (Item 7) The composition according to item 1, wherein the plurality of epitopes comprises one or more epitopes selected from SEQ ID NO: 1, 6, 8-10, 14-16, 20, 25-28, 32-34, 46-56, 60-62, 66-75, 82-85 or 87. (Item 8) The composition according to any one of items 1-7, wherein the plurality of epitopes are a plurality of contiguous epitopes. (Item 9) The composition according to item 8, wherein the contiguous epitopes further comprise a linker between one or more of the epitope sequences. (Item 10) The composition according to any one of items 1-7, further comprising an additional isolated and purified plasmid comprising an additional nucleotide sequence encoding an additional polypeptide, wherein the additional polypeptide comprises one or more epitopes having at least 90% sequence identity to an amino acid sequence selected from SEQ ID NO: 1, 6, 8-10, 14-16, 20, 25-28, 32-34, 46-56, 60-62, 66-75, 82-85 or 87. (Item 11) The composition according to any one of items 1-7, further comprising an additional isolated and purified plasmid comprising an additional nucleotide sequence encoding an additional polypeptide, wherein the additional polypeptide comprises a plurality of epitopes selected from SEQ ID NO: 1, 6, 8-10, 14-16, 20, 25-28, 32-34, 46-56, 60-62, 66-75, 82-85 or 87. (Item 12) The composition according to any one of items 1, 10 or 11, wherein the sequences of the polypeptide and the additional polypeptide are different. (Item 13) The composition according to item 1, wherein the isolated and purified plasmid further comprises a first nucleotide sequence encoding a first epitope of a first antigen expressed by cells associated with breast cancer. (Item 14) The composition according to item 1 or 13, further comprising a second nucleotide sequence encoding a second epitope of a second antigen expressed by cells associated with breast cancer. (Item 15) The composition according to any one of items 1, 13 or 14, wherein the first nucleotide sequence and the second nucleotide sequence are located on one or more isolated and purified plasmids. (Item 16) The composition according to any one of items 1 or 13 - 15, wherein the first epitope and the second epitope are independently selected from portions of the HIF-1α peptide comprising at least 90% sequence identity to the amino acid sequences selected from SEQ ID NOs: 82 - 84. (Item 17) The composition according to any one of items 1 or 13 - 15, wherein the first and second epitopes are independently selected from CD105, Yb-1, SOX-2, CDH3 or MDM2, and the first nucleotide sequence and the second nucleotide sequence are located on one or more isolated and purified plasmids. (Item 18) The nucleic acid sequence encoding the epitope of the peptide CD105 is a) a nucleotide sequence having at least 90% sequence identity to a nucleotide sequence selected from SEQ ID NOs: 2 - 5 and b) a nucleotide sequence encoding an amino acid sequence having at least 90% sequence identity to an amino acid sequence selected from SEQ ID NOs: 1, 6 or 8 - 10 The composition according to any one of items 1, 13 - 15 or 17, selected from the group consisting of. (Item 19) The nucleic acid sequence encoding the epitope of the peptide Yb-1 is a) A nucleotide sequence having at least 90% sequence identity to a nucleotide sequence selected from SEQ ID NOs: 11 - 12 and b) A nucleotide sequence encoding an amino acid sequence having at least 90% sequence identity to an amino acid sequence selected from SEQ ID NOs: 14 - 16 The composition according to any one of items 1, 13 - 15 or 17, selected from the group consisting of. (Item 20) The nucleic acid sequence encoding the epitope of the peptide SOX - 2 is a) A nucleotide sequence having at least 90% sequence identity to a nucleotide sequence selected from SEQ ID NOs: 17 - 18 and b) A nucleotide sequence encoding an amino acid sequence having at least 90% sequence identity to SEQ ID NO: 20 The composition according to any one of items 1, 13 - 15 or 17, selected from the group consisting of. (Item 21) The nucleic acid sequence encoding the epitope of the peptide CDH3 is a) A nucleotide sequence having at least 90% sequence identity to a nucleotide sequence selected from SEQ ID NOs: 21 - 24 and b) A nucleotide sequence encoding an amino acid sequence having at least 90% sequence identity to an amino acid sequence selected from SEQ ID NOs: 25 - 28 The composition according to any one of items 1, 13 - 15 or 17, selected from the group consisting of. (Item 22) The nucleic acid sequence encoding the epitope of the peptide MDM2 is a) A nucleotide sequence having at least 90% sequence identity to a nucleotide sequence selected from SEQ ID NOs: 29 - 31 and b) A nucleotide sequence encoding an amino acid sequence having at least 90% sequence identity to an amino acid sequence selected from SEQ ID NOs: 32 - 34 The composition according to any one of items 1, 13 - 15 or 17, selected from the group consisting of. (Item 23) A nucleic acid sequence encoding a fusion peptide of 5 epitopes is a) a nucleotide sequence having at least 90% sequence identity to a nucleotide sequence selected from SEQ ID NOs: 35 to 38 and b) a nucleotide sequence encoding an amino acid sequence having at least 90% sequence identity to an amino acid sequence selected from SEQ ID NOs: 39 to 42 The composition according to any one of items 1, 13 to 15 or 17, selected from the group consisting of (Item 24) The first and the second epitopes are independently selected from IGFBP-2, HER-2 or IGF-1R, and the first nucleotide sequence and the second nucleotide sequence are located in one or more isolated and purified plasmids. The composition according to any one of items 1 or 13 to 15 (Item 25) The nucleic acid sequence encoding the epitope of the peptide IGFBP-2 is a) a nucleotide sequence having at least 90% sequence identity to a nucleotide sequence selected from SEQ ID NOs: 43 to 45 and b) a nucleotide sequence encoding an amino acid sequence having at least 90% sequence identity to an amino acid sequence selected from SEQ ID NOs: 46 to 56 The composition according to any one of items 1, 13 to 15 or 24, selected from the group consisting of (Item 26) The nucleic acid sequence encoding the epitope of the peptide HER-2 is a) a nucleotide sequence having at least 90% sequence identity to a nucleotide sequence selected from SEQ ID NOs: 57 to 59 and b) a nucleotide sequence encoding an amino acid sequence having at least 90% sequence identity to an amino acid sequence selected from SEQ ID NOs: 60 to 62 The composition according to any one of items 1, 13 to 15 or 24, selected from the group consisting of (Item 27) The nucleic acid sequence encoding the epitope of the peptide IGF-1R is a) A nucleotide sequence having at least 90% sequence identity to a nucleotide sequence selected from SEQ ID NOs: 63 to 65 and b) A nucleotide sequence encoding an amino acid sequence having at least 90% sequence identity to an amino acid sequence selected from SEQ ID NOs: 66 to 75 The composition according to any one of items 1, 13 to 15 or 24, selected from the group consisting of (Item 28) A nucleic acid sequence encoding a fusion protein of three epitopes, a) A nucleotide sequence having at least 90% sequence identity to a nucleotide sequence selected from SEQ ID NOs: 76 to 78 and b) A nucleotide sequence encoding an amino acid sequence having at least 90% sequence identity to an amino acid sequence selected from SEQ ID NOs: 79 to 81 The composition according to any one of items 1, 13 to 15 or 24, selected from the group consisting of (Item 29) The composition according to any one of items 1, 13 to 15 or 17 to 23, comprising a first and a second epitope independently selected from CD105, Yb-1, SOX-2, CDH3 or MDM2. (Item 30) The composition according to any one of items 1, 13 to 15 or 17 to 23, wherein the composition further comprises a third epitope, and the first, second and third epitopes are independently selected from CD105, Yb-1, SOX-2, CDH3 or MDM2. (Item 31) The composition according to any one of items 1, 13 to 15 or 17 to 23, wherein the composition further comprises third and fourth epitopes, and the first, second, third and fourth epitopes are independently selected from CD105, Yb-1, SOX-2, CDH3 or MDM2. (Item 32) The composition further comprises third, fourth, and fifth epitopes, and the first, second, third, fourth, and fifth epitopes are independently selected from CD105, Yb-1, SOX-2, CDH3, or MDM2, and the composition according to any one of items 1, 13 to 15, or 17 to 23. (Item 33) The composition according to any one of items 1, 13 to 15, or 24 to 28, comprising first and second epitopes independently selected from IGFBP2, HER-2, or IGF-1R. (Item 34) The composition according to any one of items 1, 13 to 15, or 24 to 28, wherein the composition further comprises a third epitope, and the first, second, and third epitopes are independently selected from IGFBP2, HER-2, or IGF-1R. (Item 35) The composition according to any of the preceding items, wherein the composition further comprises a third epitope, a fourth epitope, or a fifth epitope, and the first, second, third, fourth, or fifth epitopes are independently selected from CD105, Yb-1, SOX-2, CDH3, MDM2, HIF-1α, survivin, IGFBP2, HER-2, or IGF-1R. (Item 36) The composition according to any of the preceding items, wherein the first and second nucleic acid sequences are located in a first isolated and purified plasmid. (Item 37) The composition according to any of the preceding items, wherein the second nucleic acid sequence is located in a second isolated and purified plasmid. (Item 38) The composition according to any of the preceding items, wherein the first and second nucleic acid sequences are purified to at least 70% purity. (Item 39) The composition according to any of the preceding items, wherein at least the first isolated and purified plasmid is contained within a pharmaceutical composition. (Item 40) The composition according to any one of the preceding items, wherein the pharmaceutical composition further comprises a pharmaceutical carrier, an adjuvant, or a combination thereof. (Item 41) The composition according to any one of the preceding items, further comprising an adjuvant and a pharmaceutically acceptable carrier. (Item 42) The composition according to any one of the preceding items, wherein the first and the second nucleic acid sequences are located in the first isolated and purified plasmid and are separated by the sequence of a linker nucleic acid. (Item 43) The composition according to any one of the preceding items, wherein the first nucleic acid sequence is adjacent to the second nucleic acid sequence in the first isolated and purified plasmid. (Item 44) a) a first epitope of a first antigen expressed by cells associated with breast cancer or ovarian cancer, b) a second epitope of a second antigen expressed by cells associated with breast cancer or ovarian cancer and a composition comprising wherein the first and the second epitopes independently comprise at least 90% sequence identity to an amino acid sequence selected from SEQ ID NO: 1, 6, 8-10, 14-16, 20, 25-28, 32-34, 46-56, 60-62, 66-75, 82-85 or 87. (Item 45) The composition according to item 44, wherein the first and the second epitopes are derived from HIF-1α. (Item 46) The composition according to item 44 or 45, wherein at least the first epitope of the peptide HIF-1α is selected from the group consisting of amino acid sequences having at least 90% sequence identity to an amino acid sequence selected from SEQ ID NO: 82-84. (Item 47) The composition according to item 44, wherein the first and the second epitopes are independently selected from CD105, Yb-1, SOX-2, CDH3 or MDM2. (Item 48) The composition according to item 44 or 47, wherein at least the first epitope of the peptide CD105 is selected from the group consisting of amino acid sequences having at least 90% sequence identity to the amino acid sequences selected from SEQ ID NO: 1, 6, or 8-10. (Item 49) The composition according to item 44 or 47, wherein at least the first epitope of the peptide Yb-1 is selected from the group consisting of amino acid sequences having at least 90% sequence identity to the amino acid sequences selected from SEQ ID NO: 14-16. (Item 50) The composition according to item 44 or 47, wherein at least the first epitope of the peptide SOX-2 is selected from the group consisting of amino acid sequences having at least 90% sequence identity to SEQ ID NO: 20. (Item 51) The composition according to item 44 or 47, wherein at least the first epitope of the peptide CDH3 is selected from the group consisting of amino acid sequences having at least 90% sequence identity to the amino acid sequences selected from SEQ ID NO: 25-28. (Item 52) The composition according to item 44 or 47, wherein at least the first epitope of the peptide MDM-2 is selected from the group consisting of amino acid sequences having at least 90% sequence identity to the amino acid sequences selected from SEQ ID NO: 32-34. (Item 53) The composition according to item 44 or 47, wherein the amino acid sequence of the fusion peptide of five epitopes is selected from the group consisting of amino acid sequences having at least 90% sequence identity to the amino acid sequences selected from SEQ ID NO: 39-42. (Item 54) The composition according to item 44, wherein the first and the second epitopes are independently selected from IGFBP-2, HER-2, or IGF-1R, and the first nucleotide sequence and the second nucleotide sequence are located on one or more plasmids. (Item 55) The composition according to item 44 or 54, wherein at least the first epitope of the peptide IGFBP-2 is selected from the group consisting of amino acid sequences having at least 90% sequence identity to the amino acid sequences selected from SEQ ID NOs: 46 to 56. (Item 56) (Item 57) The composition according to item 44 or 54, wherein at least the first epitope of the peptide HER-2 is selected from the group consisting of nucleotide sequences encoding amino acid sequences having at least 90% sequence identity to the amino acid sequences selected from SEQ ID NOs: 60 to 62. The composition according to item 44 or 54, wherein the nucleic acid sequence encoding the epitope of the peptide IGF-1R is selected from the group consisting of amino acid sequences having at least 90% sequence identity to the amino acid sequences selected from SEQ ID NOs: 66 to 75. (Item 58) The composition according to item 44 or 54, wherein the nucleic acid sequence encoding the fusion protein of three epitopes is selected from the group consisting of amino acid sequences having at least 90% sequence identity to the amino acid sequences selected from SEQ ID NOs: 79 to 81. (Item 59) The composition according to any one of items 44 or 47 to 53, comprising first and second epitopes independently selected from CD105, Yb-1, SOX-2, CDH3 or MDM2. (Item 60) The composition according to any one of items 44, 47 to 53 or 59, wherein the composition further comprises a third epitope, and the first, second and third epitopes are independently selected from CD105, Yb-1, SOX-2, CDH3 or MDM2. (Item 61) The composition according to any one of items 44, 47 to 53, 59 or 60, wherein the composition further comprises third and fourth epitopes, and the first, second, third and fourth epitopes are independently selected from CD105, Yb-1, SOX-2, CDH3 or MDM2. (Item 62) The composition further comprises third, fourth and fifth epitopes, and the first, second, third, fourth and fifth epitopes are independently selected from CD105, Yb-1, SOX-2, CDH3 or MDM2, and the composition according to any one of items 44, 47 to 53 or 59 to 61. (Item 63) The composition according to any one of items 44 or 47 to 53, comprising first and second epitopes independently selected from IGFBP2, HER-2 or IGF-1R. (Item 64) The composition further comprises a third epitope, and the first, second and third epitopes are independently selected from IGFBP2, HER-2 or IGF-1R, and the composition according to any one of items 44, 47 to 53 or 63. (Item 65) The composition further comprises a third epitope, a fourth epitope or a fifth epitope, and the first, second, third, fourth or fifth epitopes are independently selected from CD105, Yb-1, SOX-2, CDH3, MDM2, HIF-1α, survivin, IGFBP2, HER-2 or IGF-1R, and the composition according to any one of items 44 to 64. (Item 66) The composition according to any one of items 44 to 65, wherein at least the first epitope is contained in the pharmaceutical composition. (Item 67) The composition according to any one of items 44 to 66, wherein the pharmaceutical composition further comprises a pharmaceutical carrier, an adjuvant or a combination thereof. (Item 68) The composition according to any one of items 44 to 67, further comprising an adjuvant and a pharmaceutical carrier. (Item 69) The composition according to any one of items 44 to 68, wherein the amino acid sequences of the first and second epitopes are separated by a linker amino acid sequence. (Item 70) The composition according to any one of items 44 to 69, wherein the amino acid sequence of the first epitope is adjacent to the amino acid sequence of the second epitope. (Item 71) The composition according to any one of items 44 to 70, wherein the cancer is breast cancer. (Item 72) The composition according to any one of the preceding items, which is formulated for administration to a subject. (Item 73) The composition according to any one of the preceding items, wherein the cells related to the breast cancer are selected from breast cells expressing atypia characteristics, pre-tumor breast cells, breast cancer cells, pre-invasive breast cancer cells, breast cancer stem cells, epithelial cells, mesenchymal cells, stromal cells, or combinations thereof. (Item 74) The composition according to any one of the preceding items, which is effective in inducing an immune response in a subject. (Item 75) The composition according to any one of the preceding items, which is effective in eliminating a large number of cells related to breast cancer in a subject. (Item 76) The composition according to any one of the preceding items, which can be used for preventing the growth of cells related to breast cancer in a subject. (Item 77) The composition according to any one of the preceding items, wherein the immune response is a type 1 immune response. (Item 78) The composition according to any one of the preceding items, wherein the immune response is characterized by the ratio of type I cytokine production to type II cytokine production exceeding 1. (Item 79) The composition according to any one of the preceding items, wherein the immune response is characterized by the ratio of type I cytokine production to type II cytokine production less than 1. (Item 80) The composition according to any one of the preceding items, wherein the immune response is characterized by the ratio of IFNγ production to IL-10 production exceeding 1. (Item 81) The composition according to any one of the preceding items, wherein the immune response is characterized by a ratio of IFNγ production to IL-10 production of less than 1. (Item 82) The composition according to any one of the preceding items, wherein the adjuvant is GM-CSF. (Item 83) A method of administering the composition according to any one of the preceding items to a subject. (Item 84) The method according to item 83, wherein the subject is in need thereof. (Item 85) A method for preventing breast cancer in a subject, the method comprising administering to the subject the composition according to any one of items 1 to 82. (Item 86) A method for treating breast cancer in a subject, the method comprising administering to the subject the composition according to items 1 to 82. (Item 87) The method according to item 85 or 86, wherein the step of administering further comprises delivering at least one dose of the composition according to items 1 to 82 to the subject. (Item 88) The method according to item 85 or 86, wherein the step of administering further comprises delivering the composition according to items 1 to 82 to the subject by subcutaneous injection, intradermal injection, intramuscular injection, intravenous injection, topical application or inhalation. (Item 89) The method according to item 85 or 86, wherein the subject is selected from the group consisting of a human having breast cancer, a mouse having breast cancer and a rat having breast cancer. (Item 90) The method according to item 85 or 86, wherein the subject is selected from the group consisting of a human without breast cancer, a mouse without breast cancer and a rat without breast cancer. (Item 91) An isolated and purified plasmid comprising at least one nucleotide sequence encoding a polypeptide comprising at least 90% sequence identity to an epitope sequence selected from SEQ ID NOs: 82 to 84. (Item 92) The isolated and purified plasmid according to item 91, wherein the isolated and purified plasmid contains a set of two or more nucleotide sequences, and each of the two or more nucleotide sequences independently encodes a polypeptide containing at least 90% sequence identity selected from SEQ ID NOs: 82 to 84. (Item 93) The isolated and purified plasmid according to item 91, wherein the isolated and purified plasmid contains a set of two or more nucleotide sequences, each of the two or more nucleotide sequences encodes a polypeptide containing at least 90% sequence identity selected from SEQ ID NOs: 82 to 84, and each of the nucleotides is not identical within the set of the two or more nucleotide sequences. (Item 94) An isolated and purified plasmid containing at least one nucleotide sequence encoding a polypeptide containing at least 90% sequence identity to an epitope sequence selected from SEQ ID NOs: 1, 6, 8 to 10, 14 to 16, 20, 25 to 28 or 32 to 34. (Item 95) The isolated and purified plasmid according to item 94, wherein the isolated and purified plasmid contains a set of two or more nucleotide sequences, and each of the two or more nucleotide sequences independently encodes a polypeptide containing at least 90% sequence identity selected from SEQ ID NOs: 1, 6, 8 to 10, 14 to 16, 20, 25 to 28 or 32 to 34. (Item 96) The isolated and purified plasmid according to item 94, wherein the isolated and purified plasmid contains a set of two or more nucleotide sequences, each of the two or more nucleotide sequences encodes a polypeptide containing at least 90% sequence identity selected from SEQ ID NOs: 1, 6, 8 to 10, 14 to 16, 20, 25 to 28 or 32 to 34, and each of the nucleotides is not identical within the set of the two or more nucleotide sequences. (Item 97) An isolated and purified plasmid comprising at least one nucleotide sequence encoding a polypeptide having at least 90% sequence identity to an epitope sequence selected from SEQ ID NOs: 46-56, 60-62, or 66-75. (Item 98) The isolated and purified plasmid according to item 97, wherein the isolated and purified plasmid comprises a set of two or more nucleotide sequences, and each of the two or more nucleotide sequences independently encodes a polypeptide having at least 90% sequence identity to an epitope sequence selected from SEQ ID NOs: 46-56, 60-62, or 66-75. (Item 99) The isolated and purified plasmid according to item 97, wherein the isolated and purified plasmid comprises a set of two or more nucleotide sequences, each of the two or more nucleotide sequences encodes a polypeptide having at least 90% sequence identity to an epitope sequence selected from SEQ ID NOs: 46-56, 60-62, or 66-75, and each of the nucleotides is not identical within the set of two or more nucleotide sequences. (Item 100) A kit for preparing the composition according to any one of items 1-82, the kit comprising instructions for preparing the composition. (Item 101) A kit for administering the composition according to any one of items 1-82 to a subject, the kit comprising instructions for administering the composition. (Item 102) a) A plasmid comprising at least one nucleotide sequence encoding a polypeptide having at least 70% sequence identity to an epitope sequence selected from SEQ ID NOs: 54, 73, 85, or 87, and b) An excipient and a composition comprising the same. (Item 103) The composition according to item 102, wherein the plasmid comprises at least one nucleotide sequence encoding a polypeptide comprising at least 80% sequence identity to an epitope sequence selected from SEQ ID NO: 54, 73, 85 or 87. (Item 104) The composition according to item 102, wherein the plasmid comprises at least one nucleotide sequence encoding a polypeptide comprising at least 90% sequence identity to an epitope sequence selected from SEQ ID NO: 54, 73, 85 or 87. (Item 105) The composition according to item 102, wherein the plasmid comprises at least one nucleotide sequence encoding a polypeptide comprising at least 95% sequence identity to an epitope sequence selected from SEQ ID NO: 54, 73, 85 or 87. (Item 106) The composition according to item 102, wherein the plasmid comprises at least one nucleotide sequence encoding a polypeptide comprising 100% sequence identity to an epitope sequence selected from SEQ ID NO: 54, 73, 85 or 87. (Item 107) The composition according to item 102, wherein the plasmid comprises at least one nucleotide sequence encoding a polypeptide consisting of 100% sequence identity to the full length of an epitope sequence selected from SEQ ID NO: 54, 73, 85 or 87. (Item 108) The composition according to any one of items 102 to 107, wherein the at least one nucleotide sequence encodes a polypeptide comprising at least 70% sequence identity to at least 20 consecutive amino acids of SEQ ID NO: 85. (Item 109) The composition according to any one of items 102 to 107, wherein the at least one nucleotide sequence encodes a polypeptide comprising at least 80% sequence identity to at least 20 consecutive amino acids of SEQ ID NO: 85. (Item 110) The composition according to any one of items 102 to 107, wherein the at least one nucleotide sequence encodes a polypeptide having at least 90% sequence identity to at least 20 consecutive amino acids of SEQ ID NO: 85. (Item 111) The composition according to any one of items 102 to 107, wherein the at least one nucleotide sequence encodes a polypeptide having at least 95% sequence identity to at least 20 consecutive amino acids of SEQ ID NO: 85. (Item 112) The composition according to any one of items 102 to 107, wherein the at least one nucleotide sequence encodes a polypeptide having at least 100% sequence identity to at least 20 consecutive amino acids of SEQ ID NO: 85. (Item 113) The composition according to any one of items v, wherein the at least one nucleotide sequence encodes a polypeptide having at least 70% sequence identity to at least 60 consecutive amino acids of SEQ ID NO: 87. (Item 114) The composition according to any one of items 102 to 107, wherein the at least one nucleotide sequence encodes a polypeptide having at least 80% sequence identity to at least 60 consecutive amino acids of SEQ ID NO: 87. (Item 115) The composition according to any one of items 102 to 107, wherein the at least one nucleotide sequence encodes a polypeptide having at least 90% sequence identity to at least 60 consecutive amino acids of SEQ ID NO: 87. (Item 116) The composition according to any one of items 102 to 107, wherein the at least one nucleotide sequence encodes a polypeptide having at least 95% sequence identity to at least 60 consecutive amino acids of SEQ ID NO: 87. (Item 117) The composition according to any one of items 102 to 107, wherein the at least one nucleotide sequence encodes a polypeptide having at least 100% sequence identity to at least 60 consecutive amino acids of SEQ ID NO: 87. (Item 118) The composition according to any one of items 102 to 117, wherein the plasmid comprises at least four nucleotide sequences. (Item 119) The composition according to item 118, wherein each of the at least four nucleotide sequences independently encodes a polypeptide having at least 70% sequence identity to an epitope sequence selected from SEQ ID NO: 54, 73, 85 or 87. (Item 120) The composition according to any one of items 102 to 119, wherein the plasmid comprises four nucleotide sequences. (Item 121) The composition according to item 120, wherein each of the four nucleotide sequences independently encodes a polypeptide having at least 70% sequence identity to an epitope sequence selected from SEQ ID NO: 54, 73, 85 or 87. (Item 122) The composition according to item 120, wherein each of the four nucleotide sequences encodes a different polypeptide. (Item 123) The composition according to item 122, wherein each of the different polypeptides has at least 70% sequence identity to an epitope sequence selected from SEQ ID NO: 54, 73, 85 and 87. (Item 124) The composition according to any one of items 102 to 117, further comprising at least one additional plasmid. (Item 125) The composition according to item 124, wherein the at least one additional plasmid comprises a nucleotide sequence encoding a polypeptide having at least 70% sequence identity to an epitope sequence selected from SEQ ID NO: 54, 73, 85 or 87. (Item 126) The composition according to any one of items 102 to 125, wherein the plasmid is an isolated and purified plasmid. (Item 127) The composition according to any one of items 102 to 126, wherein the plasmid is about 50%, about 60%, about 70%, about 80%, about 90% or about 100% pure. (Item 128) The composition according to any one of items 102 to 127, wherein the plasmid is an expression vector. (Item 129) The composition according to item 128, wherein the expression vector contains pUMVC3. (Item 130) The composition according to any one of items 102 to 129, further comprising an adjuvant. (Item 131) The composition according to item 130, wherein the adjuvant is GM-CSF. (Item 132) The composition according to any one of items 102 to 131, wherein the excipient is a pharmaceutically acceptable carrier. (Item 133) The composition according to any one of items 102 to 132, formulated for subcutaneous, intramuscular or intradermal administration. (Item 134) a) A plasmid comprising four nucleotide sequences, each of the four nucleotide sequences independently encoding a polypeptide having at least 70% sequence identity to an epitope sequence selected from SEQ ID NO: 54, 73, 85 or 87, and b) An excipient A composition comprising. (Item 135) The composition according to item 134, wherein each of the four nucleotide sequences independently encodes a polypeptide having at least 80% sequence identity to an epitope sequence selected from SEQ ID NO: 54, 73, 85 or 87. (Item 136) The composition according to item 134, wherein each of the four nucleotide sequences independently encodes a polypeptide comprising at least 90% sequence identity to an epitope sequence selected from SEQ ID NO: 54, 73, 85 or 87. (Item 137) The composition according to item 134, wherein each of the four nucleotide sequences independently encodes a polypeptide comprising at least 95% sequence identity to an epitope sequence selected from SEQ ID NO: 54, 73, 85 or 87. (Item 138) The composition according to item 134, wherein each of the four nucleotide sequences independently encodes a polypeptide comprising 100% sequence identity to an epitope sequence selected from SEQ ID NO: 54, 73, 85 or 87. (Item 139) The composition according to item 134, wherein each of the four nucleotide sequences independently encodes a polypeptide consisting of 100% sequence identity to the full length of an epitope sequence selected from SEQ ID NO: 54, 73, 85 or 87. (Item 140) The composition according to item 134, wherein each of the four nucleotide sequences encodes a different polypeptide. (Item 141) The composition according to item 134 or 140, wherein each of the different polypeptides comprises at least 70% sequence identity to an epitope sequence selected from SEQ ID NO: 54, 73, 85 or 87. (Item 142) The composition according to any one of items 134 to 141, wherein one of the four nucleotide sequences encodes a polypeptide comprising at least 70% sequence identity to at least 20 consecutive amino acids of SEQ ID NO: 85. (Item 143) The composition according to any one of items 134 to 141, wherein one of the four nucleotide sequences encodes a polypeptide comprising at least 80% sequence identity to at least 20 consecutive amino acids of SEQ ID NO: 85. (Item 144) The composition according to any one of items 134 to 141, wherein one of the four nucleotide sequences encodes a polypeptide comprising at least 90% sequence identity to at least 20 consecutive amino acids of SEQ ID NO: 85. (Item 145) The composition according to any one of items 134 to 141, wherein one of the four nucleotide sequences encodes a polypeptide comprising at least 95% sequence identity to at least 20 consecutive amino acids of SEQ ID NO: 85. (Item 146) The composition according to any one of items 134 to 141, wherein one of the four nucleotide sequences encodes a polypeptide comprising at least 100% sequence identity to at least 20 consecutive amino acids of SEQ ID NO: 85. (Item 147) The composition according to any one of items 134 to 141, wherein one of the four nucleotide sequences encodes a polypeptide comprising at least 70% sequence identity to at least 60 consecutive amino acids of SEQ ID NO: 87. (Item 148) The composition according to any one of items 134 to 141, wherein one of the four nucleotide sequences encodes a polypeptide comprising at least 80% sequence identity to at least 60 consecutive amino acids of SEQ ID NO: 87. (Item 149) The composition according to any one of items 134 to 141, wherein one of the four nucleotide sequences encodes a polypeptide comprising at least 90% sequence identity to at least 60 consecutive amino acids of SEQ ID NO: 87. (Item 150) The composition according to any one of items 134 to 141, wherein one of the four nucleotide sequences encodes a polypeptide comprising at least 95% sequence identity to at least 60 consecutive amino acids of SEQ ID NO: 87. (Item 151) The composition according to any one of items 134 to 141, wherein one of the four nucleotide sequences encodes a polypeptide having at least 100% sequence identity to at least 60 consecutive amino acids of SEQ ID NO: 87. (Item 152) The composition according to any one of items 134 to 151, wherein the four nucleotide sequences are arranged tandemly within the plasmid. (Item 153) The composition according to any one of items 134 to 152, wherein the four nucleotide sequences are separated by a linker nucleic acid sequence. (Item 154) The composition according to any one of items 134 to 153, wherein the plasmid is an isolated plasmid. (Item 155) The composition according to any one of items 134 to 154, wherein the plasmid is about 50%, about 60%, about 70%, about 80%, about 90% or about 100% pure. (Item 156) The composition according to any one of items 134 to 155, wherein the plasmid is an expression vector. (Item 157) The composition according to item 156, wherein the expression vector contains pUMVC3. (Item 158) The composition according to any one of items 134 to 157, further comprising an adjuvant. (Item 159) The composition according to item 158, wherein the adjuvant is GM-CSF. (Item 160) The composition according to any one of items 134 to 159, wherein the excipient is a pharmaceutically acceptable carrier. (Item 161) The composition according to any one of items 134 to 160, formulated for subcutaneous administration, intramuscular administration or intradermal administration. (Item 162) The composition according to any one of items 102 to 133, for the treatment of breast cancer in a subject. (Item 163) The composition according to any one of Items 102 to 133 for treating ovarian cancer in a subject. (Item 164) The composition according to any one of Items 134 to 161 for treating breast cancer in a subject. (Item 165) The composition according to any one of Items 134 to 161 for treating ovarian cancer in a subject. (Item 166) The composition according to any one of Items 162 to 165, wherein the breast cancer is recurrent or refractory breast cancer. (Item 167) The composition according to any one of Items 162 to 166, wherein the breast cancer is metastatic breast cancer. (Item 168) The composition according to any one of Items 162 to 165, wherein the ovarian cancer is recurrent or refractory ovarian cancer. (Item 169) The composition according to any one of Items 162 to 165 or 168, wherein the ovarian cancer is metastatic ovarian cancer. (Item 170) The composition according to any one of Items 162 to 169 that induces an immune response. (Item 171) The composition according to Item 170, wherein the immune response is characterized by a ratio of type I cytokine production to type II cytokine production that exceeds 1. (Item 172) The composition according to Item 170, wherein the immune response is characterized by a ratio of type I cytokine production to type II cytokine production that is less than 1. (Item 173) The composition according to Item 170, wherein the immune response is characterized by a ratio of IFN-γ production to IL-10 production that exceeds 1. (Item 174) The composition according to Item 170, wherein the immune response is characterized by a ratio of IFN-γ production to IL-10 production that is less than 1. (Item 175) A method for treating breast cancer, comprising the step of administering to a subject in need thereof the composition according to items 102 to 133. (Item 176) A method for treating ovarian cancer, comprising the step of administering to a subject in need thereof the composition according to items 102 to 133. (Item 177) A method for treating breast cancer, comprising the step of administering to a subject in need thereof the composition according to items 134 to 161. (Item 178) A method for treating ovarian cancer, comprising the step of administering to a subject in need thereof the composition according to items 134 to 161. (Item 179) The method according to any one of items 175 to 178, wherein the breast cancer is a recurrent or refractory cancer. (Item 180) The method according to any one of items 175 to 179, wherein the breast cancer is a metastatic cancer. (Item 181) The method according to any one of items 175 to 178, wherein the ovarian cancer is a recurrent or refractory ovarian cancer. (Item 182) The method according to any one of items 175 to 178 or 181, wherein the ovarian cancer is a metastatic ovarian cancer. (Item 183) The method according to any one of items 175 to 182, wherein the composition induces an immune response. (Item 184) The method according to item 183, wherein the immune response is characterized by a ratio of type I cytokine production to type II cytokine production that exceeds 1. (Item 185) The method according to item 183, wherein the immune response is characterized by a ratio of type I cytokine production to type II cytokine production that is less than 1. (Item 186) The method according to item 183, wherein the immune response is characterized by a ratio of IFN-γ production to IL-10 production that exceeds 1. (Item 187) The method according to item 183, wherein the immune response is characterized by a ratio of IFN-γ production to IL-10 production that is less than 1. (Item 188) The method according to any one of items 175 to 187, further comprising the step of administering an additional therapeutic agent. (Item 189) A method for generating an immune response in a subject having breast cancer or ovarian cancer, the method comprising administering to the subject the composition according to items 102 to 133. (Item 190) A method for generating an immune response in a subject having breast cancer or ovarian cancer, the method comprising administering to the subject the composition according to items 134 to 161. (Item 191) The method according to item 189 or 190, wherein the immune response is characterized by a ratio of type I cytokine production to type II cytokine production that exceeds 1. (Item 192) The method according to item 189 or 191, wherein the immune response is characterized by a ratio of type I cytokine production to type II cytokine production that is less than 1. (Item 193) The method according to item 189 or 190, wherein the immune response is characterized by a ratio of IFN-γ production to IL-10 production that exceeds 1. (Item 194) The method according to item 189 or 190, wherein the immune response is characterized by a ratio of IFN-γ production to IL-10 production that is less than 1. (Item 195) An isolated and purified plasmid comprising at least one nucleotide sequence encoding a polypeptide comprising at least 70% sequence identity to an epitope sequence selected from SEQ ID NO: 54, 73, 85, or 87. (Item 196) The plasmid according to item 195, wherein the at least one nucleotide sequence encodes a polypeptide comprising at least 80% sequence identity to an epitope sequence selected from SEQ ID NO: 54, 73, 85 or 87. (Item 197) The plasmid according to item 195, wherein the at least one nucleotide sequence encodes a polypeptide comprising at least 90% sequence identity to an epitope sequence selected from SEQ ID NO: 54, 73, 85 or 87. (Item 198) The plasmid according to item 195, wherein the at least one nucleotide sequence encodes a polypeptide comprising at least 95% sequence identity to an epitope sequence selected from SEQ ID NO: 54, 73, 85 or 87. (Item 199) The plasmid according to item 195, wherein the at least one nucleotide sequence encodes a polypeptide comprising at least 100% sequence identity to an epitope sequence selected from SEQ ID NO: 54, 73, 85 or 87. (Item 200) The plasmid according to item 195, wherein the at least one nucleotide sequence encodes a polypeptide consisting of 100% sequence identity to the full length of an epitope sequence selected from SEQ ID NO: 54, 73, 85 or 87. (Item 201) The plasmid according to any one of items 195 to 200, wherein the at least one nucleotide sequence encodes a polypeptide comprising at least 70% sequence identity to at least 20 consecutive amino acids of SEQ ID NO: 85. (Item 202) The plasmid according to any one of items 195 to 201, wherein the at least one nucleotide sequence encodes a polypeptide comprising at least 70% sequence identity to at least 60 consecutive amino acids of SEQ ID NO: 87. (Item 203) The plasmid according to any one of items 195 to 202, wherein the isolated plasmid comprises at least 4 nucleotide sequences. (Item 204) The plasmid according to item 203, wherein each of the at least four nucleotide sequences independently encodes a polypeptide comprising at least 70% sequence identity to an epitope sequence selected from SEQ ID NO: 54, 73, 85 or 87. (Item 205) The plasmid according to any one of items 195 to 204, wherein the isolated plasmid contains four nucleotide sequences. (Item 206) The plasmid according to item 205, wherein each of the four nucleotide sequences independently encodes a polypeptide comprising at least 70% sequence identity to an epitope sequence selected from SEQ ID NO: 54, 73, 85 or 87. (Item 207) The plasmid according to item 205, wherein each of the four nucleotide sequences encodes a different polypeptide. (Item 208) The plasmid according to item 207, wherein each of the different polypeptides comprises at least 70% sequence identity to an epitope sequence selected from SEQ ID NO: 54, 73, 85 or 87. (Item 209) The plasmid according to any one of items 207 to 208, wherein the four nucleotide sequences are arranged tandemly within the plasmid. (Item 210) The plasmid according to any one of items 207 to 209, wherein the four nucleotide sequences are separated by a linker nucleic acid sequence. (Item 211) The plasmid according to any one of items 195 to 210, which is about 50%, about 60%, about 70%, about 80%, about 90% or about 100% pure. (Item 212) The plasmid according to any one of items 195 to 211, which is an expression vector. (Item 213) The plasmid according to item 212, wherein the expression vector contains pUMVC3. (Item 214) a) A plasmid comprising a nucleotide sequence encoding a polypeptide having at least 80% sequence identity to SEQ ID NO: 89, and b) an excipient A composition comprising. (Item 215) The composition according to item 214, wherein the plasmid comprises a nucleotide sequence encoding a polypeptide having at least 90% sequence identity to SEQ ID NO: 89. (Item 216) The composition according to item 214, wherein the plasmid comprises a nucleotide sequence encoding a polypeptide having at least 95% sequence identity to SEQ ID NO: 89. (Item 217) The composition according to item 214, wherein the plasmid comprises a nucleotide sequence encoding a polypeptide having 100% sequence identity to SEQ ID NO: 89. (Item 218) The composition according to item 214, wherein the plasmid comprises a nucleotide sequence encoding a polypeptide consisting of 100% sequence identity to SEQ ID NO: 89. (Item 219) A composition comprising a polypeptide having at least 80% sequence identity to SEQ ID NO: 89. (Item 220) The composition according to item 219, wherein the polypeptide has at least 90% sequence identity to SEQ ID NO: 89. (Item 221) The composition according to item 219, wherein the polypeptide has at least 95% sequence identity to SEQ ID NO: 89. (Item 222) The composition according to item 219, wherein the polypeptide has 100% sequence identity to SEQ ID NO: 89. (Item 223) The composition according to item 219, wherein the polypeptide consists of 100% sequence identity to SEQ ID NO: 89.

[0006] In another aspect, the present disclosure includes a composition comprising a first plasmid containing a first nucleotide sequence encoding a first epitope of a first antigen, wherein the first epitope is a portion of a HIF-1α peptide and the first nucleotide sequence is located in the plasmid. In yet another aspect, the present disclosure includes a composition comprising a first plasmid containing a first nucleotide sequence encoding a first epitope of a first antigen and a second nucleotide sequence encoding a second epitope of a second antigen, wherein the first and second epitopes are portions of a HIF-1α peptide and the first nucleotide sequence and the second nucleotide sequence are located in one or more plasmids.

[0007] In some aspects, the compositions described herein include a composition comprising a first plasmid containing a first nucleotide sequence encoding a first epitope of a first antigen, wherein the first epitope is a portion of a peptide selected from CD105, Yb-1, SOX-2, CDH3, or MDM2 and the first nucleotide sequence is located in the plasmid. In other aspects, the present disclosure includes a composition comprising a first plasmid containing a first nucleotide sequence encoding a first epitope of a first antigen and a second nucleotide sequence encoding a second epitope of a second antigen, wherein the first and second epitopes are independently selected from CD105, Yb-1, SOX-2, CDH3, or MDM2 and the first nucleotide sequence and the second nucleotide sequence are located in one or more plasmids.

[0008] In some embodiments, the compositions described herein include a composition comprising a first plasmid containing a first nucleotide sequence encoding a first epitope of a first antigen, wherein the first epitope is a portion of a peptide selected from IGFBP-2, HER-2, IGF-1R, and the first nucleotide sequence is located in the plasmid. In some other embodiments, the present disclosure provides a composition comprising a first plasmid containing a first nucleotide sequence encoding a first epitope of a first antigen and a second nucleotide sequence encoding a second epitope of a second antigen, wherein the first and second epitopes are independently selected from IGFBP-2, HER-2 or IGF-1R, and the first nucleotide sequence and the second nucleotide sequence are located in one or more plasmids.

[0009] In some embodiments, the compositions described herein include a composition comprising a first epitope of a first antigen expressed by cells associated with breast cancer and a second epitope of a second antigen expressed by cells associated with breast cancer.

[0010] In other embodiments, the present disclosure provides a composition comprising at least a first epitope of a first antigen, wherein the first epitope is a portion of a peptide derived from HIF-1α. In still other embodiments, the present disclosure provides a composition comprising at least a first epitope of a first antigen and at least a second epitope of a second antigen, wherein the first and second epitopes are derived from HIF-1α.

[0011] In other embodiments, the present disclosure provides a composition comprising at least a first epitope of a first antigen, wherein the first epitope is a portion of a peptide selected from CD105, Yb-1, SOX-2, CDH3 or MDM2. In still other embodiments, the present disclosure provides a composition comprising at least a first epitope of a first antigen and at least a second epitope of a second antigen, wherein the first and second epitopes are independently selected from CD105, Yb-1, SOX-2, CDH3 or MDM2.

[0012] In some cases, the present disclosure includes a composition comprising an isolated and purified plasmid comprising a nucleotide sequence encoding a polypeptide comprising a plurality of epitopes, and an excipient. Optionally, the plurality of epitopes comprises one or more epitopes having at least 90% sequence identity to an amino acid sequence selected from SEQ ID NOs: 1, 6, 8-10, 14-16, 20, 25-28, 32-34, 46-56, 60-62, 66-75, 82-85, and 87.

[0013] Optionally, the isolated and purified plasmid can further comprise a first nucleotide sequence encoding a first epitope of a first antigen expressed by cells associated with breast cancer. In some examples, the composition further comprises a second nucleotide sequence encoding a second epitope of a second antigen expressed by cells associated with breast cancer. The first nucleotide sequence and the second nucleotide sequence can be located in one or more isolated and purified plasmids. The first epitope and the second epitope can be independently selected from a portion of the HIF-1α peptide having at least 90% sequence identity to an amino acid sequence selected from SEQ ID NOs: 82-84. The first and second epitopes can be independently selected from CD105, Yb-1, SOX-2, CDH3, or MDM2, and the first nucleotide sequence and the second nucleotide sequence are located in one or more isolated and purified plasmids. The first and second epitopes can be independently selected from IGFBP-2, HER-2, or IGF-1R, and the first nucleotide sequence and the second nucleotide sequence are located in one or more isolated and purified plasmids. The first and second nucleic acid sequences can be located in the first isolated and purified plasmid. The second nucleic acid sequence can be located in the second isolated and purified plasmid.

[0014] In some examples, the present disclosure provides a composition comprising a first epitope of a first antigen expressed by cells associated with breast cancer or ovarian cancer and a second epitope of a second antigen expressed by cells associated with breast cancer or ovarian cancer, wherein the first and second epitopes independently comprise at least 90% sequence identity to an amino acid sequence selected from SEQ ID NOs: 1, 6, 8-10, 14-16, 20, 25-28, 32-34, 46-56, 60-62, 66-75, 82-85, and 87.

[0015] In some cases, the present disclosure provides a composition comprising at least one nucleotide sequence encoding a polypeptide that comprises at least 70% sequence identity to an epitope sequence selected from SEQ ID NOs: 54, 73, 85, and 87, and an excipient.

[0016] In some examples, the present disclosure provides a composition comprising a plasmid comprising four nucleotide sequences and an excipient, wherein each of the four nucleotide sequences independently encodes a polypeptide that comprises at least 70% sequence identity to an epitope sequence selected from SEQ ID NOs: 54, 73, 85, and 87.

[0017] Optionally, the present disclosure provides a composition comprising a plasmid comprising a nucleotide sequence encoding a polypeptide that comprises at least 80% sequence identity to SEQ ID NO: 89, and an excipient.

[0018] Optionally, the present disclosure provides a composition comprising a polypeptide that comprises at least 80% sequence identity to SEQ ID NO: 89.

[0019] In some cases, methods of administering one or more of the compositions described herein to a subject are disclosed herein. Optionally, the subject may be in need of one or more of the compositions.

[0020] Optionally, there is provided a method for preventing breast cancer or ovarian cancer in a subject, the method comprising administering to the subject a composition described herein. Optionally, the cancer can be ovarian cancer. The cancer can be breast cancer. There is provided a method for preventing breast cancer in a subject, the method comprising administering to the subject a composition described herein.

[0021] Optionally, there is provided a method for treating breast cancer or ovarian cancer in a subject, the method comprising administering to the subject a composition described herein. Optionally, the cancer can be ovarian cancer. The cancer can be breast cancer. There is provided a method for treating breast cancer in a subject, the method comprising administering to the subject a composition described herein.

[0022] Optionally, the step of administering further comprises delivering at least one dose of the composition described herein to the subject. Optionally, the step of administering further comprises delivering the composition described herein to the subject by subcutaneous injection, intradermal injection, intramuscular injection, intravenous injection, topical application or inhalation.

[0023] In some cases, there is provided a method for eliciting an immune response in a subject having breast cancer or ovarian cancer, the method comprising administering to the subject a composition described herein.

[0024] The present disclosure further includes an isolated and purified plasmid comprising at least one nucleotide sequence encoding a polypeptide having at least 90% sequence identity to an epitope sequence selected from SEQ ID NOs: 82 to 84. The isolated and purified plasmid can comprise a set of two or more nucleotide sequences, each of the two or more nucleotide sequences independently encoding a polypeptide having at least 90% sequence identity to an epitope sequence selected from SEQ ID NOs: 82 to 84. The isolated and purified plasmid can comprise a set of two or more nucleotide sequences, each of the two or more nucleotide sequences encoding a polypeptide having at least 90% sequence identity to an epitope sequence selected from SEQ ID NOs: 82 to 84, and each of the nucleotides is not identical within the set of two or more nucleotide sequences.

[0025] The present disclosure can also include an isolated and purified plasmid comprising at least one nucleotide sequence encoding a polypeptide having at least 90% sequence identity to an epitope sequence selected from SEQ ID NOs: 1, 6, 8 to 10, 14 to 16, 20, 25 to 28, and 32 to 34. The isolated and purified plasmid can comprise a set of two or more nucleotide sequences, each of the two or more nucleotide sequences independently encoding a polypeptide having at least 90% sequence identity to an epitope sequence selected from SEQ ID NOs: 1, 6, 8 to 10, 14 to 16, 20, 25 to 28, and 32 to 34. The isolated and purified plasmid can comprise a set of two or more nucleotide sequences, each of the two or more nucleotide sequences encoding a polypeptide having at least 90% sequence identity to a sequence selected from SEQ ID NOs: 1, 6, 8 to 10, 14 to 16, 20, 25 to 28, and 32 to 34, and each of the nucleotides is not identical within the set of two or more nucleotide sequences.

[0026] The present disclosure can include an isolated and purified plasmid comprising at least one nucleotide sequence encoding a polypeptide having at least 90% sequence identity to an epitope sequence selected from SEQ ID NOs: 46-56, 60-62, or 66-75. The isolated and purified plasmid can include a set of two or more nucleotide sequences, each of the two or more nucleotide sequences independently encoding a polypeptide having at least 90% sequence identity to an epitope sequence selected from SEQ ID NOs: 46-56, 60-62, or 66-75. The isolated and purified plasmid can include a set of two or more nucleotide sequences, each of the two or more nucleotide sequences encoding a polypeptide having at least 90% sequence identity to an epitope sequence selected from SEQ ID NOs: 46-56, 60-62, or 66-75, and each of the nucleotides not being identical within the set of two or more nucleotide sequences.

[0027] Optionally, the present disclosure can further include an isolated and purified plasmid comprising at least one nucleotide sequence encoding a polypeptide having at least 70% sequence identity to an epitope sequence selected from SEQ ID NOs: 54, 73, 85, and 87. Incorporation by reference

[0028] All publications, patents, and patent applications mentioned herein are hereby incorporated by reference to the same extent as if each individual publication, patent, or patent application was specifically and individually indicated to be incorporated by reference.

[0029] The novel features of the present disclosure are particularly set forth in the appended claims. A better understanding of the features and advantages of the present disclosure will be obtained by reference to the following detailed description that sets forth illustrative embodiments in which the principles of the present disclosure are utilized, and to the appended drawings. BRIEF DESCRIPTION OF THE DRAWINGS

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Mode for Carrying Out the Invention

[0076] The present disclosure provides compositions of breast cancer vaccines and ovarian cancer vaccines, often for the prevention or treatment of breast cancer or ovarian cancer. The present disclosure further provides methods of administering a breast cancer vaccine or an ovarian cancer vaccine to a subject. The compositions provided herein can be used in combination with the methods provided herein for the prevention or treatment of breast cancer or ovarian cancer.

[0077] In some examples, the composition can include a nucleic acid sequence encoding an epitope of a breast cancer or ovarian cancer antigen that is capable of inducing an immunogenic response in a subject, a plasmid containing the sequences described herein, an adjuvant, a pharmaceutical carrier, and an inert chemical suitable for use with a pharmaceutical composition. The breast cancer or ovarian cancer antigen can be at least one of any antigen that can be expressed in a subject who may have or develop breast cancer or ovarian cancer. In many cases, the breast cancer or ovarian cancer antigen is expressed by tissues such as breast cancer cells, ovarian cancer cells, and / or breast cancer or ovarian cancer cancer stem cells (CSCs). CSCs can exhibit the potential for self-renewal, uncontrolled growth, and drug resistance. In some examples, CSCs can express a protein (e.g., an antigen), and for example, the level of protein (e.g., antigen) expression by CSCs can be upregulated (e.g., increased expression compared to a predetermined amount) or downregulated (e.g., decreased expression compared to a predetermined amount). In some examples, proteins upregulated by CSCs compared to normal tissues or cells can be involved in the development and / or progression of breast cancer or ovarian cancer. For example, the compositions and methods described herein can be used to identify proteins and target epitopes of antigens.

[0078] In some examples, one epitope of a breast cancer or ovarian cancer antigen can be used in the composition. In other examples, more than one epitope of a breast cancer or ovarian cancer antigen can be used in the composition. In other examples, more than two antigens, more than three, four, five, six, seven, eight, nine, ten, fifteen, twenty, twenty-five, or thirty breast cancer or ovarian cancer antigens can be used in the composition. In some examples, the antigens can be the same. In other examples, the antigens can be different. The breast cancer or ovarian cancer vaccine compositions described herein can be formulated for the prevention of breast cancer or ovarian cancer. For example, the prophylactic composition can eliminate cells (such as CSCs such as breast CSCs or ovarian CSCs) having abnormal (e.g., upregulated) expression of proteins to prevent breast cancer or ovarian cancer.

[0079] In some examples, the epitope(s) can be present on the same breast cancer or ovarian cancer antigen, or the epitope(s) can be present on different breast cancer or ovarian cancer antigens. In some examples, one epitope of a breast cancer or ovarian cancer antigen can be used in the composition. In other examples, more than one epitope of a breast cancer or ovarian cancer antigen, more than two antigens, three, four, five, six, seven, eight, nine, ten, fifteen, twenty, twenty-five, or thirty epitopes of a breast cancer or ovarian cancer antigen can be used in the composition.

[0080] The compositions and methods described herein can elicit an immune response in a subject. The immune response can be an immune response against an epitope of an antigen in the composition (e.g., a vaccine). The vaccine arms the subject's immune system so that the immune system can detect and destroy what contains the antigen of the vaccine in the subject. The compositions and methods described herein can elicit a type 1 (Th1) immune response in a subject. The Th1 immune response can include the secretion of inflammatory cytokines (e.g., IFNγ, TNFα) by a subset of immune cells (e.g., antigen-specific T cells). In some examples, the inflammatory cytokines activate another subtype of immune cells (e.g., cytotoxic T cells) that can destroy what contains the antigen in the subject.

[0081] Using the screening methods described herein for identifying epitopes and binding peptides from tumor antigens, epitopes of multiple antigens can be screened for induction of a Th1 immune response. For example, the screening method can identify an epitope from at least one tumor antigen that elicits (e.g., preferentially causes the secretion of Th1 cytokines) a Th1 response against a breast cancer or ovarian cancer antigen, including the CSC (e.g., breast CSC or ovarian CSC) antigens described herein.

[0082] In some examples, the epitopes and / or antigens used in the compositions and methods described herein can be recognized by the subject's immune system to elicit a Th1 immune response and release type I cytokines. The Th1 response can be induced by the interaction between the epitope and T cells, more specifically, the major histocompatibility complex (MHC) expressed by T cells. For example, high-affinity binding of the epitope to the MHC receptor can stimulate the Th1 response. The MHC receptor can be at least one of multiple types of MHC receptors. The MHC receptors associated with T cells can vary among individuals in a population.

[0083] The compositions described herein can include additional components in addition to the nucleic acid encoding the epitope of the antigen. In some examples, the composition can include at least one adjuvant. In some examples, the composition can include at least one pharmaceutical carrier. In some examples, the composition can include at least one inert chemical substance suitable for use with a pharmaceutical composition. In some examples, the composition can include at least one adjuvant and at least one pharmaceutical carrier. In some examples, the composition can include at least one adjuvant and at least one inert chemical substance suitable for use with a pharmaceutical composition. In some examples, the composition can include at least one inert chemical substance suitable for use with a pharmaceutical composition and a pharmaceutical carrier. In some examples, the composition can contain a plurality of adjuvants, a plurality of pharmaceutical carriers, and a plurality of inert chemical substances suitable for use with a pharmaceutical composition.

[0084] In some examples, one adjuvant can be used in the composition. In other examples, more than one adjuvant, more than two adjuvants, more than three adjuvants, more than four adjuvants, more than five adjuvants, more than six adjuvants, more than seven adjuvants, more than eight adjuvants, more than nine adjuvants, or more than ten adjuvants can be used in the composition. In some examples, one pharmaceutical carrier can be used in the composition. In other examples, more than one pharmaceutical carrier, more than two pharmaceutical carriers, more than three pharmaceutical carriers, more than four adjuvants, more than five pharmaceutical carriers, more than six pharmaceutical carriers, more than seven pharmaceutical carriers, more than eight pharmaceutical carriers, more than nine pharmaceutical carriers, or more than ten pharmaceutical carriers can be used in the composition. In some examples, one chemical substance can be used in the composition. In other examples, more than one chemical substance, more than two chemical substances, more than three chemical substances, more than four chemical substances, more than five chemical substances, more than six chemical substances, more than seven chemical substances, more than eight chemical substances, more than nine chemical substances, or more than ten chemical substances can be used in the composition.

[0085] The present disclosure further describes methods of administering a breast cancer or ovarian cancer vaccine to a subject. In some examples, the method can include constructing a plasmid-based vaccine that targets these antigens and determining whether administration of the vaccine is safe, immunogenic, and effective in preventing breast cancer development. For example, the composition can be a vaccine based on a multi-antigen Th1 polyepitope plasmid. In some examples, the method can include conducting at least one clinical trial to determine the safety and immunogenicity of the plasmid-based vaccine in subjects with breast cancer or ovarian cancer. For example, the antigen can be expressed by or associated with a CSC (e.g., a breast CSC or an ovarian CSC) and / or can be associated with the transition of cells from epithelial cells to mesenchymal cells (EMT). In some examples, the epitope of the composition can be derived from the antigen and the epitope can induce a Th1 immune response in the subject. For example, the Th1 immune response can include immune cells, often CD4+ T cells. In some examples, the composition can be a nucleic acid (e.g., a plasmid-based vaccine) that includes nucleic acids encoding more than one antigen or more than one epitope of an antigen. In some examples, the methods are used to determine whether the compositions described herein prevent the development of cancer (e.g., breast cancer or ovarian cancer) using a plurality of organisms, e.g., genetically diverse rodents (e.g., mice), including genetically similar rodents (e.g., mice), and in subjects with or without breast cancer or ovarian cancer. In some cases, the cancer can be breast cancer. In some examples, the breast cancer can be triple-negative breast cancer (TNBC). Identification of Antigens

[0086] The compositions and methods described herein include the identification and genetic manipulation of breast or ovarian cancer antigens in pharmaceutical compositions (e.g., vaccines). Any technique known to those of skill in the art may be used to identify antigens expressed by a subject having breast or ovarian cancer, but in an exemplary example, suitable antigens can be identified using the methods described herein. In some examples, the method can include the step of screening serum from a subject. In some examples, the screening can be an antibody screening. For example, the antibody being screened can be an IgG antibody. In some examples, the serum can be from a subject having breast or ovarian cancer. In other examples, the serum can be from a subject not having breast or ovarian cancer.

[0087] For example, a cancer antigen such as a breast cancer antigen or an ovarian cancer antigen can be a protein portion, a peptide portion, or a polyamino acid portion. In some examples, the portion can be a percentage of a protein, a peptide, or a polyamino acid. In some examples, the percentage can be less than 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% of the protein, peptide, or polyamino acid. In some examples, the portion can be located at the C-terminus of the protein, peptide, or polyamino acid. In other examples, the portion can be located near the C-terminus of the protein, peptide, or polyamino acid. For example, near the C-terminus can be within 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, or 50% of the length of the total protein, peptide, or polyamino acid from the central point. In some examples, the portion can be located at the N-terminus of the protein, peptide, or polyamino acid. In other examples, the portion can be located near the N-terminus of the protein, peptide, or polyamino acid. For example, near the N-terminus can be within 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, or 50% of the length of the total protein, peptide, or polyamino acid from the central point. In some examples, the portion can be located near the center of the protein, peptide, or polyamino acid. In other examples, the portion can be located near the center of the protein, peptide, or polyamino acid. For example, near the center can be within 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, or 50% of the length of the total protein, peptide, or polyamino acid from the end.

[0088] Regarding the suitability as an antigen in the compositions (e.g., vaccines) described herein, at least one antigen can be identified and screened. In some examples, one antigen can be identified and screened. In other examples, regarding suitability in a vaccine, more than one antigen can be identified and screened, more than two antigens can be identified and screened, more than three antigens can be identified and screened, more than four antigens can be identified and screened, more than five antigens can be identified and screened, more than six antigens can be identified and screened, more than seven antigens can be identified and screened, more than eight antigens can be identified and screened, more than nine antigens can be identified and screened, more than ten antigens can be identified and screened, more than eleven antigens can be identified and screened, more than twelve antigens can be identified and screened, more than thirteen antigens can be identified and screened, more than fourteen antigens can be identified and screened, more than fifteen antigens can be identified and screened, more than twenty antigens can be identified and screened, more than twenty-five antigens can be identified and screened, more than thirty antigens can be identified and screened, more than thirty-five antigens can be identified and screened, more than forty antigens can be identified and screened, more than forty-five antigens can be identified and screened, or more than fifty antigens can be identified and screened. In an exemplary example, regarding suitability in a vaccine, five antigens can be identified and screened.

[0089] Antigens screened for suitability in a vaccine can be derived from any protein detected in sera from subjects with breast cancer or ovarian cancer using screening techniques known to those of skill in the art. In some examples, screening can often be antibody screening. The protein can be any protein detected in sera from subjects with breast cancer or ovarian cancer, but in exemplary instances, proteins from which the antigen can be derived can be classified as stem cell proteins and / or EMT proteins. For example, breast cancer stem cell / EMT proteins can include SOX2, YB1, CD105, MDM2, CDH3+ / −, and HIF1α. Often, the antigen can be immunogenic in both breast cancer subjects and subjects without breast cancer. Mapping of Antigen Epitopes

[0090] The compositions and methods provided herein include mapping of at least one epitope within an antigen such that administration of the epitope to a subject results in a Th1 immune response. In some examples, the epitope can be administered as a breast cancer vaccine or an ovarian cancer vaccine. Any technique known to those of skill in the art can be used to identify an epitope that can induce a Th1 immune response in a subject, but the methods described herein can preferably be used. In some examples, the epitope can be part of the antigen (e.g., as identified above). For example, the epitope can be a peptide of an antigenic protein and / or a portion of an antigenic protein.

[0091] In some examples, the epitope can be a human leukocyte antigen (HLA) class I epitope derived from a breast cancer or ovarian cancer antigen. For example, the HLA class I epitope can include epitopes that bind to HLA-A, -B, and -C molecules. In some examples, the epitope can be a class II epitope derived from a breast cancer or ovarian cancer antigen for the development of a cancer vaccine (e.g., breast cancer or ovarian cancer). For example, the HLA class II epitope can include epitopes that bind to HLA-DP, -DM, -DOA, -DOB, -DQ, and -DR molecules. In some examples, in addition to the methods described herein, (1) determining whether the epitope binds (e.g., with high affinity) to the MHC by at least one HLA allele (e.g., HLA-DR, i.e., a universal epitope), (2) determining whether the epitope stimulates IFNγ rather than IL-10 secretion (e.g., from antigen-specific T cells), and (3) determining whether the T cell can recognize the peptide (e.g., the epitope) processed by the antigen-presenting cell (APC), i.e., whether it is a native epitope, can be used to map the epitope. In some examples, a T cell line can be used. For example, the T cell line can be an epitope-derived T cell line. In some examples, the T cell can be an exogenous T cell genetically engineered to express a chimeric antigen receptor construct that binds to the epitope with high selectivity and avidity. In some examples, the epitope can be derived from a protein (e.g., a recombinant protein). In other examples, the protein can be a native protein. In some examples, the protein can be processed endogenously. In other examples, the protein can be processed exogenously. In some examples, the protein can be processed endogenously by autologous APCs. In other examples, the protein can be processed exogenously by autologous APCs.

[0092] In all examples, the peptide is an epitope mapped from an antigen and can be identified using the methods described herein for the selection of peptide epitopes. In some examples, the epitope can be derived from a human protein that can be used directly in a peptide-based vaccine. In other examples, the epitope can be derived from a human protein and the encoding nucleic acid sequence can be incorporated into a nucleic acid construct designed to induce expression of the epitope in a subject after administration. For example, the nucleic acid construct can enable an immune response against at least one epitope to be coordinated, amplified, attenuated, suppressed or eliminated against a specific set of self-proteins. In some examples, the peptide or nucleic acid construct can be optimized for protein- or plasmid-based vaccination to induce, amplify or coordinate a Th1 immune response. In some examples, the epitope can be an extended Th1 epitope. In other examples, the peptide or nucleic acid construct can be optimized for protein- or plasmid-based vaccination to suppress, attenuate or eliminate a pathological response in a subject (e.g., human or animal) in need thereof.

[0093] In some examples, a protein, peptide or polyamino acid stimulates the secretion of IFNγ, and the peptide is located within the portion of the protein, peptide or polyamino acid. In some examples, a protein, peptide or polyamino acid inhibits the secretion of IFNγ, and the peptide is located within the portion of the protein, peptide or polyamino acid. In some examples, a protein, peptide or polyamino acid stimulates the secretion of IL-10, and the peptide is located within the portion of the protein, peptide or polyamino acid. In some examples, a protein, peptide or polyamino acid inhibits the secretion of IL-10, and the peptide is located within the portion of the protein, peptide or polyamino acid. In some examples, the peptide can stimulate the secretion of IFNγ and inhibit the secretion of IL-10. In other examples, the peptide can stimulate the secretion of IL-10 and inhibit the secretion of IFNγ. In some examples, the peptide can stimulate the secretion of IFNγ and stimulate the secretion of IL-10. In other examples, the peptide can inhibit the secretion of IL-10 and inhibit the secretion of IFNγ.

[0094] In some examples, the amino acids that make up the peptide can be adjusted so that the desired effect of the peptide on IFNγ secretion and / or the desired effect of the peptide on IL-10 secretion can be achieved. For example, if the peptide is to stimulate only the secretion of IFNγ, the length of the peptide can be shortened to eliminate the amino acids that stimulate IL-10 secretion, thereby adjusting the peptide that stimulates the secretion of both IFNγ and IL-10.

[0095] In some examples, the identified epitopes can be included in the vaccine composition of the extended epitope vaccine. In some examples, the extended epitope can be a 40 - 80mer peptide. In an exemplary example, either a nucleic acid sequence or a peptide sequence is juxtaposed for the construction of the extended epitope sequence. The juxtaposition of selected peptides within the parent protein (e.g., within 10 amino acids of each other) enables the construction of in - tandem extended epitopes that may contain tolerogenic and / or inhibitory epitopes. For example, the in - tandem extended epitope can contain a short intervening <10 amino acid sequence. Any of these peptides and / or extended epitopes (embodied either as the peptide itself or as the corresponding nucleic acid construct) can be optimized, either alone or in any combination, into a protein - or plasmid - based vaccination that specifically induces, amplifies or tunes a protective immune response, or suppresses, attenuates or eliminates a pathological response, in a subject (human or animal) in need thereof.

[0096] In some examples, an epitope can be the length of an amino acid. In some examples, the epitope can be less than 5 amino acids, less than 10 amino acids, less than 15 amino acids, less than 20 amino acids, less than 25 amino acids, less than 30 amino acids, less than 35 amino acids, less than 40 amino acids, less than 45 amino acids, less than 50 amino acids, less than 55 amino acids, less than 60 amino acids, less than 70 amino acids, less than 75 amino acids, less than 80 amino acids, less than 85 amino acids, less than 90 amino acids, less than 95 amino acids, less than 100 amino acids, less than 110 amino acids, less than 120 amino acids, less than 130 amino acids, less than 140 amino acids, less than 150 amino acids, less than 160 amino acids, less than 170 amino acids, less than 180 amino acids, less than 190 amino acids, less than 200 amino acids, less than 210 amino acids, less than 220 amino acids, less than 230 amino acids, less than 240 amino acids, less than 250 amino acids, less than 260 amino acids, less than 270 amino acids, less than 280 amino acids, less than 290 amino acids, less than 300 amino acids, less than 350 amino acids, less than 400 amino acids, less than 450 amino acids or less than 500 amino acids.

[0097] In some cases, the present disclosure provides a composition comprising an isolated and purified plasmid comprising a nucleotide sequence encoding a polypeptide comprising a plurality of epitopes, and an excipient. In some cases, the plurality of epitopes comprises one or more epitopes having at least 90% sequence identity to an amino acid sequence selected from SEQ ID NOs: 1, 6, 8-10, 14-16, 20, 25-28, 32-34, 46-56, 60-62, 66-75, 82-85, and 87. In some examples, the plurality of epitopes comprises one or more epitopes having at least 90% sequence identity to an amino acid sequence selected from SEQ ID NOs: 82-84. In some examples, the plurality of epitopes comprises one or more epitopes having at least 90% sequence identity to an amino acid sequence selected from SEQ ID NOs: 1, 6, 8-10, 14-16, 20, 25-28, or 32-34. In some examples, the plurality of epitopes comprises one or more epitopes having at least 90% sequence identity to an amino acid sequence selected from SEQ ID NOs: 46-56, 60-62, or 66-75. In some examples, the plurality of epitopes comprises one or more epitopes having at least 90% sequence identity to an amino acid sequence selected from SEQ ID NOs: 54, 73, 85, and 87. In some examples, the plurality of epitopes comprises one or more epitopes selected from SEQ ID NOs: 1, 6, 8-10, 14-16, 20, 25-28, 32-34, 46-56, 60-62, 66-75, 82-85, and 87.

[0098] In some examples, the plurality of epitopes are a plurality of contiguous epitopes. In some examples, the contiguous epitopes further comprise a linker between one or more of the epitope sequences. In some examples, the amino acid sequences of the first and second epitopes are separated by the sequence of linker amino acids. In some examples, the amino acid sequence of the first epitope is adjacent to the amino acid sequence of the second epitope.

[0099] In some examples, the composition further comprises an additional isolated and purified plasmid comprising an additional nucleotide sequence encoding an additional polypeptide, the additional polypeptide comprising one or more epitopes comprising a plurality of epitopes having at least 90% sequence identity to an amino acid sequence selected from SEQ ID NOs: 1, 6, 8-10, 14-16, 20, 25-28, 32-34, 46-56, 60-62, 66-75, 82-85, and 87. Optionally, the composition further comprises an additional isolated and purified plasmid comprising an additional nucleotide sequence encoding an additional polypeptide, the additional polypeptide comprising a plurality of epitopes selected from SEQ ID NOs: 1, 6, 8-10, 14-16, 20, 25-28, 32-34, 46-56, 60-62, 66-75, 82-85, and 87. In some examples, the sequences of the polypeptide and the additional polypeptide are different.

[0100] In some examples, the immune response is a type 1 immune response. In some examples, the immune response is characterized by a ratio of type I cytokine production to type II cytokine production greater than 1. In some examples, the immune response is characterized by a ratio of type I cytokine production to type II cytokine production less than 1. In some examples, the immune response is characterized by a ratio of IFNγ production to IL-10 production greater than 1. In some examples, the immune response is characterized by a ratio of IFNγ production to IL-10 production less than 1.

[0101] In some examples, the composition is administered to a subject. In some examples, the subject is in need of administration of the composition. In some examples, the composition is effective in inducing an immune response in a subject. In some examples, the composition is effective in eliminating a plurality of cells associated with breast cancer or ovarian cancer in a subject. In some examples, the composition can be used to prevent the growth of cells associated with breast cancer or ovarian cancer in a subject.

[0102] In some examples, the cancer is breast cancer. In some cases, the breast cancer is recurrent or refractory or metastatic breast cancer. In some cases, the cancer is ovarian cancer. In some examples, the ovarian cancer is recurrent or refractory or metastatic ovarian cancer.

[0103] In some examples, at least the first epitope is contained within a pharmaceutical composition. In some examples, at least the first epitope is contained within a pharmaceutical composition that further comprises a pharmaceutical carrier. In some examples, at least the first epitope is contained within a pharmaceutical composition that further comprises a pharmaceutical carrier and an adjuvant. In some examples, at least the first epitope is contained within a pharmaceutical composition that further comprises an adjuvant. In some examples, the composition further comprises an adjuvant and a pharmaceutical carrier. In some examples, the adjuvant is GM-CSF.

[0104] The present disclosure also provides a kit for preparing the compositions described herein, the kit comprising instructions for preparing the compositions. The present disclosure also provides a kit for administering the compositions described herein, the kit comprising instructions for administering the compositions. Composition comprising an epitope for a breast cancer vaccine

[0105] The compositions described herein comprise a first plasmid comprising a first nucleotide sequence encoding a first epitope of a first antigen expressed by cells associated with breast cancer, and a second nucleotide sequence encoding a second epitope of a second antigen expressed by cells associated with breast cancer, wherein the first nucleotide sequence and the second nucleotide sequence are located on one or more plasmids. In some examples, the composition can comprise nucleic acids encoding epitopes from the following proteins, CD105, HIF1α, MDM2, Yb1, SOX-2, HER-2, IGFBP2, IGF-1R, CDH3, and survivin.

[0106] In some examples, the composition is a composition comprising a first plasmid comprising a first nucleotide sequence encoding a first epitope of a first antigen, wherein the first epitope is a portion of a peptide selected from CD105, Yb-1, SOX-2, CDH3 or MDM2, and the first nucleotide sequence can comprise a composition located in the plasmid. In other examples, the composition is a composition comprising a first plasmid comprising a first nucleotide sequence encoding a first epitope of a first antigen and a second nucleotide sequence encoding a second epitope of a second antigen, wherein the first and second epitopes are independently selected from CD105, Yb-1, SOX-2, CDH3 or MDM2, and the first nucleotide sequence and the second nucleotide sequence can comprise a composition located in one or more plasmids.

[0107] In some examples, the composition can include nucleic acids encoding epitopes from the following proteins, CD105, MDM2, Yb-1, SOX-2, and CDH3. In some examples, the composition has at least 90% sequence identity to the nucleotide sequence of CAGAACGGCACCTGGCCCCGCGAGGTGCTGCTGGTGCTGTCCGTGAACTCCTCCGTGTTCCTGCACCTACAGGCCCTGGGCATCCCCCTGCACCTGGCCTACAACTCCTCCCTGGTGACCTTCCAGGAGCCCCCCGGCGTGAACACCACCGAGCTG (SEQ ID NO: 2); at least 90% sequence identity to the nucleotide sequence of ACCGTGTTCATGCGCCTGAACATCATCTCCCCCGACCTGTCCGGCTGCACCTCCAAGGGCCTGGTGCTGCCCGCCGTGCTGGGCATCACCTTCGGCGCCTTCCTGATCGGCGCCCTGCTGACCGCCGCCCTGTGGTACATCTACTCCCACACCCGCTCCCCCTCCAAGCGCGAGCCCGTGGTGGCCGTGGCCGCCCCCGCCTCCTCCGAGTCCTCCTCCACCAACCACTCCATCGGCTCCACCCAGTCCACCCCCTGCTCCACCTCCTCCATGGCC (SEQ ID NO: 3); at least 90% sequence identity to the nucleotide sequence of ACCGTGTCCATGCGCCTGAACATCGTGTCCCCCGACCTGTCCGGCAAGGGCCTGGTGCTGCCCTCCGTGCTGGGCATCACCTTCGGCGCCTTCCTGATCGGCGCCCTGCTGACCGCCGCCCTGTGGTACATCTACTCCCACACCCGCGGCCCCTCCAAGCGCGAGCCCGTGGTGGCCGTGGCCGCCCCCGCCTCCTCCGAGTCCTCCTCCACCAACCACTCCATCGGCTCCACCCAGTCCACCCCCTGCTCCACCTCCTCCATGGCC (SEQ ID NO: 4); at least 90% sequence identity to A nucleotide sequence having at least 90% sequence identity to the nucleotide sequence: ACCGTGTCCATGCGCCTGAACATCGTGTCCCCCGACCTGTCCGGCAAGGGCCTGGTGCTGCCCTCCGTGCTGGGCATCACCTTCGGCGCCTTCCTGATCGGCGCCCTGCTGACCGCCGCCCTGTGGTACATCTACTCCCACACCCGCGCCCCCTCCAAGCGCGAGCCCGTGGTGGCCGTGGCCGCCCCCGCCTCCTCCGAGTCCTCCTCCACCAACCACTCCATCGGCTCCACCCAGTCCACCCCCTGCTCCACCTCCTCCATGGCC (SEQ ID NO: 5); a nucleotide sequence encoding an amino acid sequence having at least 90% sequence identity to the amino acid sequence: EARMLNASIVASFVELPL (SEQ ID NO: 6); a nucleotide sequence encoding an amino acid sequence having at least 90% sequence identity to the amino acid sequence: QNGTWPREVLLVLSVNSSVFLHLQALGIPLHLAYNSSLVTFQEPPGVNTTEL (SEQ ID NO: 7); a nucleotide sequence encoding an amino acid sequence having at least 90% sequence identity to the amino acid sequence: TVFMRLNIISPDLSGCTSKGLVLPAVLGITFGAFLIGALLTAALWYIYSHTRSPSKREPVVAVAAPASSESSSTNHSIGSTQSTPCSTSSMA (SEQ ID NO: 8); a nucleotide sequence encoding an amino acid sequence having at least 90% sequence identity to the amino acid sequence: TVSMRLNIVSPDLSGKGLVLPSVLGITFGAFLIGALLTAALWYIYSHTRGPSKREPVVAVAAPASSESSSTNHSIGSTQSTPCSTSSMA (SEQ ID NO: 9); or a nucleotide sequence encoding an amino acid sequence having at least 90% sequence identity to the amino acid sequence: TVSMRLNIVSPDLSGKGLVLPSVLGITFGAFLIGALLTAALWYIYSHTRAPSKREPVVAVAAPASSESSSTNHSIGSTQSTPCSTSSMA (SEQ ID NO: 10). It can contain a nucleic acid sequence encoding an epitope of peptide CD105 selected from the group consisting of these nucleotide sequences.In some examples, the composition comprises a nucleotide sequence having at least 90% sequence identity to the nucleotide sequence of GGAGTGCCAGTGCAGGGCTCCAAGTACGCTGCCGACCGCAACCACTACCGCCGCTACCCACGCCGTCGCGGCCCACCCCGCAACTACCAGCAGAAC (SEQ ID NO: 11); a nucleotide sequence having at least 90% sequence identity to the nucleotide sequence of GGCGTGCCCGTGCAGGGCTCCAAGTACGCCGCCGACCGCAACCACTACCGCCGCTACCCCCGCCGCCGCGGCCCCCCCCGCAACTACCAGCAGAAC (SEQ ID NO: 12); a nucleotide sequence having at least 90% sequence identity to the nucleotide sequence of GGCGTGCCCGTGCAGGGCTCCAAGTACGCCGCCGACCGCAACCACTACCGCCGCTACCCCCGCCGCCGCGGCCCCCCCCGCAACTACCAGCAGAAC (SEQ ID NO: 13); a nucleotide sequence encoding an amino acid sequence having at least 90% sequence identity to the amino acid sequence of EDVFVHQTAIKKNNPRK (SEQ ID NO: 14); a nucleotide sequence encoding an amino acid sequence having at least 90% sequence identity to the amino acid sequence of YRRNFNYRRRRPEN (SEQ ID NO: 15); or a nucleotide sequence encoding an amino acid sequence having at least 90% sequence identity to the amino acid sequence of GVPVQGSKYAADRNHYRRYPRRRGPPRNYQQN (SEQ ID NO: 16), and can comprise a nucleic acid sequence encoding an epitope of peptide Yb-1 selected from the group consisting of. In some examples, the composition has at least 90% sequence to the nucleotide sequence of GGCCTCAATGCGCACGGCGCAGCGCAGATGCAGCCCATGCACCGCTACGACGTGAGCGCCCTGCAGTACAACTCCATGACCAGCTCGCAGACCTACATGAACGGCTCGCCCACCTACAGCATGTCCTACTCGCAGCAGGGCACCCCTGGCATGGCTCTTGGCTCCATGGGTTCGGTG (SEQ ID NO: 17). Nucleotide sequence having identity; nucleotides of GGCCTGAACGCCCACGGCGCCGCCCAGATGCAGCCCATGCACCGCTACGACGTGTCCGCCCTGCAGTACAACTCCATGACCTCCTCCCAGACCTACATGAACGGCTCCCCCACCTACTCCATGTCCTACTCCCAGCAGGGCACCCCCGGCATGGCCCTGGGCTCCATGGGCTCCGTG (SEQ ID NO: 18) Nucleotide sequence having at least 90% sequence identity to the sequence; GGCCTGAACGCCCACGGCGCCGCCCAGATGCAGCCCATGCACCGCTACGACGTGTCCGCCCTGCAGTACAACTCCATGACCTCCTCCCAGACCTACATGAACGGCTCCCCCACCTACTCCATGTCCTACTCCCAGCAGGGCACCCCCGGCATGGCCCTGGGCTCCATGGGCTCCGTG A nucleotide sequence having at least 90% sequence identity to the nucleotide sequence of (Accession No. 19); or a nucleotide sequence encoding an amino acid sequence having at least 90% sequence identity to the amino acid sequence of GLNAHGAAQMQPMHRYDVSALQYNSMTSSQTYMNGSPTYSMSYSQQGTPGMALGSMGSV (Accession No. 20). It can include a nucleic acid sequence encoding an epitope of the peptide SOX-2 selected from the group consisting of. In some examples, the composition has at least 90% sequence identity to the nucleotide sequence of AGGTCACTGAAGGAAAGGAATCCATTGAAAATCTTCCCATCCAAACGTATCTTACGAAGACACAAGAGAGATTGGGTGGTTGCTCCAATATCTGTCCCTGAAAATGGCAAGGGTCCCTTCCCACAGAGACTGAATCAGCTCAAGTCTAATAAAGATAGAGACACCAAGATTTTCTACAGCATCACGGGGCCGGGTGCAGACAGCCCACCTGAGGGTGTCTTCGCTGTAGAGAAGGAGACA (Accession No. 21); at least 90% sequence identity to the nucleotide sequence of TTGAAAATCTTCCCATCCAAACGTATCTTACGAAGACACAAGAGAGATTGGGTGGTTGCTCCAATATCTGTCCCTGAAAATGGCAAGGGTCCCTTCCCACAGAGACTGAATCAGCTCAAGTCTAATAAAGATAGAGACACCAAGATTTTCTACAGCATCACGGGGCCGGGTGCAGACAGCCCACCTGAGGGTGTCTTCGCTGTAGAGAAGGAGACA (Accession No. 22);A nucleotide sequence having at least 90% sequence identity to the nucleotide sequence of GCCATGCACTCCCCCCCCACCCGCATCCTGCGCCGCCGCAAGCGCGAGTGGGTGATGCCCCCCATCTTCGTGCCCGAGAACGGCAAGGGCCCCTTCCCCCAGCGCCTGAACCAGCTGAAGTCCAACAAGGACCGCGGCACCAAGATCTTCTACTCCATCACCGGCCCCGGCGCCGACTCCCCCCCCGAGGGCGTGTTCACCATCGAGAAGGAGTCC (SEQ ID NO: 23); A nucleotide sequence having at least 90% sequence identity to the nucleotide sequence of GTGATGAACTCCCCCCCCTCCCGCATCCTGCGCCGCCGCAAGCGCGAGTGGGTGATGCCCCCCATCTCCGTGCCCGAGAACGGCAAGGGCCCCTTCCCCCAGCGCCTGAACCAGCTGAAGTCCAACAAGGACCGCGGCACCAAGCTGTTCTACTCCATCACCGGCCCCGGCGCCGACTCCCCCCCCGAGGGCGTGTTCACCATCGAGAAGGAGACC (SEQ ID NO: 24); A nucleotide sequence encoding an amino acid sequence having at least 90% sequence identity to the amino acid sequence of RSLKERNPLKIFPSKRILRRHKRDWVVAPISVPENGKGPFPQRLNQLKSNKDRDTKIFYSITGPGADSPPEGVFAVEKET (SEQ ID NO: 25); A nucleotide sequence encoding an amino acid sequence having at least 90% sequence identity to the amino acid sequence of LKIFPSKRILRRHKRDWVVAPISVPENGKGPFPQRLNQLKSNKDRDTKIFYSITGPGADSPPEGVFAVEKET (SEQ ID NO: 26); A nucleotide sequence encoding an amino acid sequence having at least 90% sequence identity to the amino acid sequence of AMHSPPTRILRRRKREWVMPPIFVPENGKGPFPQRLNQLKSNKDRGTKIFYSITGPGADSPPEGVFTIEKES (SEQ ID NO: 27);Or a nucleic acid sequence encoding an epitope of peptide CDH3 selected from the group consisting of nucleotide sequences encoding an amino acid sequence having at least 90% sequence identity to the amino acid sequence of VMNSPPSRILRRRKREWVMPPISVPENGKGPFPQRLNQLKSNKDRGTKLFYSITGPGADSPPEGVFTIEKET (SEQ ID NO: 28). In some examples, the composition is ACCTACACCATGAAGGAGGTGCTGTTCTACCTGGGCCAGTACATCATGACCAAGCGCCTGTACGACGAGAAGCAGCAGCACATCGTGTACTGCTCCAACGACCTGCTGGGCGACCTGTTCGGCGTGCCCTCCTTCTCCGTGAAGGAGCACCGCAAaATCTACACCATGATCTACCGCAACCTGGTGGTGGTGAACCAGCAGGAGTCCTCCGACTCCGGCACCTCCGTGTCC (SEQ ID NO: 29) nucleotides; A nucleotide sequence having at least 90% sequence identity to the d array; a nucleotide sequence having at least 90% sequence identity to the nucleotide sequence of ACCTACACCATGAAGGAGATCATCTTCTACATCGGCCAGTACATCATGACCAAGCGCCTGTACGACGAGAAGCAGCAGCACATCGTGTACTGCTCCAACGACCTGCTGGGCGACGTGTTCGGCGTGCCCTCCTTCTCCGTGAAGGAGCACCGCAAGATCTACGCCATGATCTACCGCAACCTGGTGGCCGTGTCCCAGCAGGACTCCGGCACCTCCCTGTCC (SEQ ID NO: 30); a nucleotide sequence having at least 90% sequence identity to the nucleotide sequence of ATCTACACCATGAAGGAGATCATCTTCTACATCGGCCAGTACATCATGACCAAGCGCCTGTACGACGAGAAGCAGCAGCACATCGTGTACTGCTCCAACGACCTGCTGGGCGACGTGTTCGGCGTGCCCTCCTTCTCCGTGAAGGAGCACCGCAAGATCTACGCCATGATCTACCGCAACCTGGTGGTGGTGTCCCAGCAGGACTCCGGCACCTCCCCCTCC (SEQ ID NO: 31); a nucleotide sequence encoding an amino acid sequence having at least 90% sequence identity to the amino acid sequence of TYTMKEVLFYLGQYIMTKRLYDEKQQHIVYCSNDLLGDLFGVPSFSVKEHRKIYTMIYRNLVVVNQQESSDSGTSV (SEQ ID NO: 32); a nucleotide sequence encoding an amino acid sequence having at least 90% sequence identity to the amino acid sequence of TYTMKEIIFYIGQYIMTKRLYDEKQQHIVYCSNDLLGDVFGVPSFSVKEHRKIYAMIYRNLVAVSQQDSGTSLS (SEQ ID NO: 33);Or a nucleic acid sequence encoding an epitope of peptide MDM2 selected from the group consisting of nucleotide sequences encoding an amino acid sequence having at least 90% sequence identity to the amino acid sequence of IYTMKEIIFYIGQYIMTKRLYDEKQQHIVYCSNDLLGDVFGVPSFSVKEHRKIYAMIYRNLVVVSQQDSGTSPS (SEQ ID NO: 34) can be included.;

[0108] In an exemplary example, the composition is,ATGGCGGTACCCATGCAACTGTCCTGCTCTAGACAGAACGGCACCTGGCCCCGCGAGGTGCTGCTGGTGCTGTCCGTGAACTCCTCCGTGTTCCTGCACCTACAGGCCCTGGGCATCCCCCTGCACCTGGCCTACAACTCCTCCCTGGTGACCTTCCAGGAGCCCCCCGGCGTGAACACCACCGAGCTGAGATCCACCGGTGGAGTGCCAGTGCAGGGCTCCAAGTACGCTGCCGACCGCAACCACTACCGCCGCTACCCACGCCGTCGCGGCCCACCCCGCAACTACCAGCAGAACACGCGTGGCCTCAATGCGCACGGCGCAGCGCAGATGCAGCCCATGCACCGCTACGACGTGAGCGCCCTGCAGTACAACTCCATGACCAGCTCGCAGACCTACATGAACGGCTCGCCCACCTACAGCATGTCCTACTCGCAGCAGGGCACCCCTGGCATGGCTCTTGGCTCCATGGGTTCGGTGAGATCCCAATTGAGGTCACTGAAGGAAAGGAATCCATTGAAAATCTTCCCATCCAAACGTATCTTACGAAGACACAAGAGAGATTGGGTGGTTGCTCCAATATCTGTCCCTGAAAATGGCAAGGGTCCCTTCCCACAGAGACTGAATCAGCTCAAGTCTAATAAAGATAGAGACACCAAGATTTTCTACAGCATCACGGGGCCGGGTGCAGACAGCCCACCTGAGGGTGTCTTCGCTGTAGAGAAGGAGACAAGATCCGCCGGCGAAACCTACACCATGAAGGAGGTGCTGTTCTACCTGGGCCAGTACATCATGACCAAGCGCCTGTACGACGAGAAGCAGCAGCACATCGTGTACTGCTCCAACGACCTGCTGGGCGACCTGTTCGGCGTGCCCTCCTTCTCCGTGAAGGAGCACCGCAAAATCTACACCATGATCTACCGCAACCTGGTGGTGGTGAACCAGCAGGAGTCCTCCGACTCCGGCACCTCCGTGTCCAGATCTTAG (SEQ ID NO: 35) nucleotides,A nucleotide sequence having at least 90% sequence identity to the nucleotide sequence encoding an amino acid sequence having at least 90% sequence identity to the amino acid sequence MAVPMQLSCSRQNGTWPREVLLVLSVNSSVFLHLQALGIPLHLAYNSSLVTFQEPPGVNTTELRSTGGVPVQGSKYAADRNHYRRYPRRRGPPRNYQQNTRGLNAHGAAQMQPMHRYDVSALQYNSMTSSQTYMNGSPTYSMSYSQQGTPGMALGSMGSVRSQLRSLKERNPLKIFPSKRILRRHKRDWVVAPISVPENGKGPFPQRLNQLKSNKDRDTKIFYSITGPGADSPPEGVFAVEKETRSAGETYTMKEVLFYLGQYIMTKRLYDEKQQHIVYCSNDLLGDLFGVPSFSVKEHRKIYTMIYRNLVVVNQQESSDSGTSVSRS (SEQ ID NO: 39); A nucleotide sequence encoding an amino acid sequence having at least 90% sequence identity to the amino acid sequence MAVPMTVFMRLNIISPDLSGCTSKGLVLPAVLGITFGAFLIGALLTAALWYIYSHTRSPSKREPVVAVAAPASSESSSTNHSIGSTQSTPCSTSSMATGGVPVQGSKYAADRNHYRRYPRRRGPPRNYQQNTRGLNAHGAAQMQPMHRYDVSALQYNSMTSSQTYMNGSPTYSMSYSQQGTPGMALGSMGSVRSQLLKIFPSKRILRRHKRDWVVAPISVPENGKGPFPQRLNQLKSNKDRDTKIFYSITGPGADSPPEGVFAVEKETRSAGETYTMKEVLFYLGQYIMTKRLYDEKQQHIVYCSNDLLGDLFGVPSFSVKEHRKIYTMIYRNLVVVNQQESSDSGTSVSRS (SEQ ID NO: 40);A nucleotide sequence encoding an amino acid sequence having at least 90% sequence identity to the amino acid sequence of MAVPMTVSMRLNIVSPDLSGKGLVLPSVLGITFGAFLIGALLTAALWYIYSHTRGPSKREPVVAVAAPASSESSSTNHSIGSTQSTPCSTSSMATGGVPVQGSKYAADRNHYRRYPRRRGPPRNYQQNTRGLNAHGAAQMQPMHRYDVSALQYNSMTSSQTYMNGSPTYSMSYSQQGTPGMALGSMGSVRSQLAMHSPPTRILRRRKREWVMPPIFVPENGKGPFPQRLNQLKSNKDRGTKIFYSITGPGADSPPEGVFTIEKESRSAGETYTMKEIIFYIGQYIMTKRLYDEKQQHIVYCSNDLLGDVFGVPSFSVKEHRKIYAMIYRNLVAVSQQDSGTSLSRS (SEQ ID NO: 41); or a nucleotide sequence encoding an amino acid sequence having at least 90% sequence identity to the amino acid sequence of MAVPMTVSMRLNIVSPDLSGKGLVLPSVLGITFGAFLIGALLTAALWYIYSHTRAPSKREPVVAVAAPASSESSSTNHSIGSTQSTPCSTSSMATGGVPVQGSKYAADRNHYRRYPRRRGPPRNYQQNTRGLNAHGAAQMQPMHRYDVSALQYNSMTSSQTYMNGSPTYSMSYSQQGTPGMALGSMGSVRSQLVMNSPPSRILRRRKREWVMPPISVPENGKGPFPQRLNQLKSNKDRGTKLFYSITGPGADSPPEGVFTIEKETRSAGEIYTMKEIIFYIGQYIMTKRLYDEKQQHIVYCSNDLLGDVFGVPSFSVKEHRKIYAMIYRNLVVVSQQDSGTSPSRS (SEQ ID NO: 42). The nucleic acid sequence can include a nucleic acid sequence encoding a fusion peptide of 5 epitopes selected from the group consisting of;

[0109] In some examples, the composition can include nucleic acids encoding epitopes derived from the following proteins, HER-2, IGFBP2, and IGF-1R. In some examples, the composition can include a first plasmid comprising a first nucleotide sequence encoding a first epitope of a first antigen, the first epitope being a portion of a peptide selected from IGFBP-2, HER-2, IGF-1R, and the first nucleotide sequence being located in the plasmid. In some examples, the composition can include a first plasmid comprising a first nucleotide sequence encoding a first epitope of a first antigen and a second nucleotide sequence encoding a second epitope of a second antigen, the first and second epitopes being independently selected from IGFBP-2, HER-2, or IGF-1R, and the first nucleotide sequence and the second nucleotide sequence being located in one or more plasmids.In one example, the composition has a nucleotide sequence having at least 90% sequence identity to the nucleotide sequence of ATGCTGCCGAGAGTGGGCTGCCCCGCGCTGCCGCTGCCGCCGCCGCCGCTGCTGCCGCTGCTGCCGCTGCTGCTGCTGCTACTGGGCGCGAGTGGCGGCGGCGGCGGGGCGCGCGCGGAGGTGCTGTTCCGCTGCCCGCCCTGCACACCCGAGCGCCTGGCCGCCTGCGGGCCCCCGCCGGTTGCGCCGCCCGCCGCGGTGGCCGCAGTGGCCGGAGGCGCCCGCATGCCATGCGCGGAGCTCGTCCGGGAGCCGGGCTGCGGCTGCTGCTCGGTGTGCGCCCGGCTGGAGGGCGAGGCGTGCGGCGTCTACACCCCGCGCTGCGGCCAGGGGCTGCGCTGCTATCCCCACCCGGGCTCCGAGCTGCCCCTGCAGGCGCTGGTCATGGGCGAGGGCACTTGTGAGAAGCGCCGGGACGCCGAGTATGGCGCCAGCCCGGAGCAGGTTGCAGACAATGGCGATGACCACTCAGAAGGAGGCCTGGTGGAG (SEQ ID NO: 43); the nucleo of ATGCTGCCCCGCCTGGGCGGCCCCGCCCTGCCCCTGCTGCTGCCCTCCCTGCTGCTGCTGCTGCTGCTGGGCGCCGGCGGCTGCGGCCCCGGCGTGCGCGCCGAGGTGCTGTTCCGCTGCCCCCCCTGCACCCCCGAGCGCCTGGCCGCCTGCGGCCCCCCCCCCGACGCCCCCTGCGCCGAGCTGGTGCGCGAGCCCGGCTGCGGCTGCTGCTCCGTGTGCGCCCGCCAGGAGGGCGAGGCCTGCGGCGTGTACATCCCCCGCTGCGCCCAGACCCTGCGCTGCTACCCCAACCCCGGCTCCGAGCTGCCCCTGAAGGCCCTGGTGACCGGCGCCGGCACCTGCGAGAAGCGCCGCGTGGGCACCACCCCCCAGCAGGTGGCCGACTCCGACGACGACCACTCCGAGGGCGGCCTGGTGGAG (SEQ ID NO: 44). A nucleotide sequence having at least 90% sequence identity to the oligonucleotide sequence; at least 90% sequence identity to the nucleotide sequence of ATGCTGCCCCGCCTGGGCGGCCCCGCCCTGCCCCTGCTGCTGCCCTCCCTGCTGCTGCTGCTGCTGCTGGGCGCCGGCGGCTGCGGCCCCGGCGTGCGCGCCGAGGTGCTGTTCCGCTGCCCCCCCTGCACCCCCGAGCGCCTGGCCGCCTGCGGCCCCCCCCCCGACGCCCCCTGCGCCGAGCTGGTGCGCGAGCCCGGCTGCGGCTGCTGCTCCGTGTGCGCCCGCCAGGAGGGCGAGGCCTGCGGCGTGTACATCCCCCGCTGCGCCCAGACCCTGCGCTGCTACCCCAACCCCGGCTCCGAGCTGCCCCTGAAGGCCCTGGTGACCGGCGCCGGCACCTGCGAGAAGCGCCGCGTGGGCACCACCCCCCAGCAGGTGGCCGACTCCGAGGACGACCACTCCGAGGGCGGCCTGGTGGAG (SEQ ID NO: 45) The nucleotide sequence; TMRRLLQETELVEPLTPSGAMPNQAQMRILKETELRKVKVLGSGAFGTVYKGIWIPDGENVKIPVAIKVLRENTSPKANKEILDEAYVMAGVGSPYVSRLLGICLTSTVQLVTQLMPYGCLLDHVRENRGRLGSQDLLNWCMQIAKGMSYLEDVRLVHRDLAARNVLVKSPNHVKITDFGLARLLDIDETEYHADGGKVPIKWMALESILRRRFTHQSDVWSYGVTVWELMTFGAKPYDGIPAREIPDLLEKGERLPQPPICTIDVYMIMVKCWMIDSECRPRFRELVSEFSRMARDPQRFVVIQNEDLAPGAGGMVHHRHRSSSPLPAARPAGATLERPKTLSPGKNGVVKDVFAFGGAVENPEYL (SEQ ID NO: 60) of A nucleotide sequence encoding an amino acid sequence having at least 90% sequence identity to the amino acid sequence; a nucleotide sequence encoding an amino acid sequence having at least 90% sequence identity to the amino acid sequence of TMRRLLQETELVEPLTPSGAVPNQAQMRILKETELRKLKVLGSGAFGTVYKGIWIPDGENVKIPVAIKVLRENTSPKANKEILDEAYVMAGVGSPYVSRLLGICLTSTVQLVTQLMPYGCLLDHVREHRGRLGSQDLLNWCVQIAKGMSYLEEVRLVHRDLAARNVLVKSPNHVKITDFGLARLLDIDETEYHADGGKVPIKWMALESILRRRFTHQSDVWSYGVTVWELMTFGAKPYDGIPAREIPDLLEKGERLPQPPICTIDVYMIMVKCWMIDSECRPRFRELVSEFSRMARDPQRFVVIQNEDLALGTGSTAHRRHRSSSPPPPIRPAGATLERPKTLSPGKNGVVKDVFAFGGAVENPEYL (SEQ ID NO: 61); and TMRRLLQETELVEPLTPSGAMPNQAQMRILKETELRKVKVLGSGAFGTVYKGIWIPDGENVKIPVAIKVLRENTSPKANKEILDEAYVMAGVGSPYVSRLLGICLTSTVQLVTQLMPYGCLLDHVREHRGRLGSQDLLNWCVQIAKGMSYLEDVRLVHRDLAARNVLVKSPNHVKITDFGLARLLDIDETEYHADGGKVPIKWMALESILRRRFTHQSDVWSYGVTVWELMTFGAKPYDGIPAREIPDLLEKGERLPQPPICTIDVYMIMVKCWMIDSECRPRFRELVSEFSRMARDPQRFVVIQNEDLTPGTGSTAHRRHRSSSPLPPVRPAGATLERPKTLSPGKNGVVKDVFAFGGAVENPEYL (SEQ ID NO: 6 A nucleotide sequence having at least 90% sequence identity to the nucleotide sequence; ATGGCGGTACCAATGCTGCCCCGCCTGGGCGGCCCCGCCCTGCCCCTGCTGCTGCCCTCCCTGCTGCTGCTGCTGCTGCTGGGCGCCGGCGGCTGCGGCCCCGGCGTGCGCGCCGAGGTGCTGTTCCGCTGCCCCCCCTGCACCCCCGAGCGCCTGGCCGCCTGCGGCCCCCCCCCC

[0110] In some examples, the composition can include first and second epitopes independently selected from CD105, Yb-1, SOX-2, CDH3, or MDM2. In some examples, the composition can include a third epitope, and the first, second, and third epitopes are independently selected from CD105, Yb-1, SOX-2, CDH3, or MDM2. In some examples, the composition can include third and fourth epitopes, and the first, second, third, and fourth epitopes are independently selected from CD105, Yb-1, SOX-2, CDH3, or MDM2. In some examples, the composition can include third, fourth, and fifth epitopes, and the first, second, third, fourth, and fifth epitopes are independently selected from CD105, Yb-1, SOX-2, CDH3, or MDM2.

[0111] In some examples, the composition can include first and second epitopes independently selected from IGFBP2, HER-2, or IGF-1R. In some examples, the composition can include a third epitope, and the first, second, and third epitopes are independently selected from IGFBP2, HER-2, or IGF-1R.

[0112] In some examples, the composition can be administered to a subject. In some examples, the subject is in need of administration of the composition. In some examples, the composition is effective in inducing an immune response in the subject. In some examples, the composition is effective in eliminating a number of cells associated with breast cancer in the subject. In some examples, the composition can be used to prevent the growth of cells associated with breast cancer in the subject.

[0113] In some examples, the first and second nucleic acid sequences are located on a first plasmid. In some examples, the second nucleic acid sequence is located on a second plasmid.

[0114] In some examples, the cells associated with breast cancer are selected from breast cells that exhibit atypia, pre-tumor breast cells, breast cancer cells, pre-invasive breast cancer cells, breast cancer stem cells, epithelial cells, mesenchymal cells, stromal cells, or combinations thereof.

[0115] In some examples, the first and second nucleic acid sequences are purified to at least 70% purity. In some examples, the first and second nucleic acid sequences are located on a first plasmid and separated by the sequence of a linker nucleic acid. In some examples, the first nucleic acid sequence is adjacent to the second nucleic acid sequence in the first plasmid.

[0116] In some examples, at least the first plasmid is contained within a pharmaceutical composition. In some examples, at least the first plasmid is contained within a pharmaceutical composition that further comprises a pharmaceutical carrier. In some examples, at least the first plasmid is contained within a pharmaceutical composition that further comprises a pharmaceutical carrier and an adjuvant. In some examples, at least the first plasmid is contained within a pharmaceutical composition that further comprises an adjuvant. In some examples, the composition further comprises an adjuvant and a pharmaceutically acceptable carrier. In some examples, the adjuvant is GM-CSF.

[0117] In some examples, the subject is selected from a human having breast cancer, a mouse having breast cancer, or a rat having breast cancer. In some examples, the subject is selected from a human without breast cancer, a mouse without breast cancer, or a rat without breast cancer.

[0118] In some examples, the immune response is a type 1 immune response. In some examples, the first nucleic acid sequence is a species selected from human, mouse, or rat. In some examples, the second nucleic acid sequence is a species selected from human, mouse, or rat. In some examples, the immune response is characterized by the ratio of type I cytokine production to type II cytokine production greater than 1. In some examples, the immune response is characterized by the ratio of type I cytokine production to type II cytokine production less than 1. In some examples, the immune response is characterized by the ratio of IFNγ production to IL-10 production greater than 1. In some examples, the immune response is characterized by the ratio of IFNγ production to IL-10 production less than 1.

[0119] In some examples, the composition is a composition comprising a first plasmid comprising a first nucleotide sequence encoding a first epitope of a first antigen, wherein the first epitope is a portion of the HIF-1α peptide, and the first nucleotide sequence is located in the plasmid. In other examples, the composition is a composition comprising a first plasmid comprising a first nucleotide sequence encoding a first epitope of a first antigen and a second nucleotide sequence encoding a second epitope of a second antigen, wherein the first and second epitopes are portions of the HIF-1α peptide, and the first nucleotide sequence and the second nucleotide sequence are located in one or more plasmids.

[0120] Nucleic acid sequences encoding epitopes from the following proteins, CD105, HIF1α, MDM2, Yb1, SOX-2, HER-2, IGFBP2, IGF-1R, and CDH3, can be different from those described herein. In some examples, nucleic acid sequences that are homologous to the nucleic acid sequences disclosed herein by more than 95%, 90%, 85%, 80%, 75%, 70%, 65%, 60%, 55%, or more than 50% can be used in the compositions described herein.

[0121] In some examples, the compositions described herein can include a composition comprising a first epitope of a first antigen expressed by cells associated with breast cancer and a second epitope of a second antigen expressed by cells associated with breast cancer.

[0122] In some examples, the composition can comprise at least a first epitope of a first antigen, wherein the first epitope is a portion of a peptide selected from CD105, Yb-1, SOX-2, CDH3, or MDM2. In some examples, the composition can comprise at least a first epitope of a first antigen and at least a second epitope of a second antigen, wherein the first and second epitopes are independently selected from CD105, Yb-1, SOX-2, CDH3, or MDM2. In some examples, at least a first epitope of the peptide CD105 is an amino acid sequence having at least 90% sequence identity to the amino acid sequence of EARMLNASIVASFVELPL (SEQ ID NO: 6); an amino acid sequence having at least 90% sequence identity to the amino acid sequence of QNGTWPREVLLVLSVNSSVFLHLQALGIPLHLAYNSSLVTFQEPPGVNTTEL (SEQ ID NO: 1); an amino acid sequence having at least 90% sequence identity to the amino acid sequence of TVFMRLNIISPDLSGCTSKGLVLPAVLGITFGAFLIGALLTAALWYIYSHTRSPSKREPVVAVAAPASSESSSTNHSIGSTQSTPCSTSSMA (SEQ ID NO: 8); an amino acid sequence having at least 90% sequence identity to the amino acid sequence of TVSMRLNIVSPDLSGKGLVLPSVLGITFGAFLIGALLTAALWYIYSHTRGPSKREPVVAVAAPASSESSSTNHSIGSTQSTPCSTSSMA (SEQ ID NO: 9); or an amino acid sequence having at least 90% sequence identity to the amino acid sequence of TVSMRLNIVSPDLSGKGLVLPSVLGITFGAFLIGALLTAALWYIYSHTRAPSKREPVVAVAAPASSESSSTNHSIGSTQSTPCSTSSMA (SEQ ID NO: 10), and is selected from the group consisting of these amino acid sequences.In some examples, at least the first epitope of peptide Yb-1 is an amino acid sequence having at least 90% sequence identity to the amino acid sequence of EDVFVHQTAIKKNNPRK (SEQ ID NO: 14); an amino acid sequence having at least 90% sequence identity to the amino acid sequence of YRRNFNYRRRRPEN (SEQ ID NO: 15); or an amino acid sequence having at least 90% sequence identity to the amino acid sequence of GVPVQGSKYAADRNHYRRYPRRRGPPRNYQQN (SEQ ID NO: 16), and is selected from the group consisting thereof. In some examples, at least the first epitope of peptide SOX-2 is selected from the group consisting of amino acid sequences having at least 90% sequence identity to the amino acid sequence of GLNAHGAAQMQPMHRYDVSALQYNSMTSSQTYMNGSPTYSMSYSQQGTPGMALGSMGSV (SEQ ID NO: 20). In some examples, at least the first epitope of peptide CDH3 is an amino acid sequence having at least 90% sequence identity to the amino acid sequence of RSLKERNPLKIFPSKRILRRHKRDWVVAPISVPENGKGPFPQRLNQLKSNKDRDTKIFYSITGPGADSPPEGVFAVEKET (SEQ ID NO: 25); an amino acid sequence having at least 90% sequence identity to the amino acid sequence of LKIFPSKRILRRHKRDWVVAPISVPENGKGPFPQRLNQLKSNKDRDTKIFYSITGPGADSPPEGVFAVEKET (SEQ ID NO: 26); an amino acid sequence having at least 90% sequence identity to the amino acid sequence of AMHSPPTRILRRRKREWVMPPIFVPENGKGPFPQRLNQLKSNKDRGTKIFYSITGPGADSPPEGVFTIEKES (SEQ ID NO: 27); or an amino acid sequence having at least 90% sequence identity to the amino acid sequence of VMNSPPSRILRRRKREWVMPPISVPENGKGPFPQRLNQLKSNKDRGTKLFYSITGPGADSPPEGVFTIEKET (SEQ ID NO: 28), and is selected from the group consisting thereof.In some examples, at least the first epitope of the peptide MDM-2 consists of an amino acid sequence having at least 90% sequence identity to the amino acid sequence of TYTMKEVLFYLGQYIMTKRLYDEKQQHIVYCSNDLLGDLFGVPSFSVKEHRKIYTMIYRNLVVVNQQESSDSGTSV (SEQ ID NO: 32); an amino acid sequence having at least 90% sequence identity to the amino acid sequence of TYTMKEIIFYIGQYIMTKRLYDEKQQHIVYCSNDLLGDVFGVPSFSVKEHRKIYAMIYRNLVAVSQQDSGTSLS (SEQ ID NO: 33); or an amino acid sequence having at least 90% sequence identity to the amino acid sequence of IYTMKEIIFYIGQYIMTKRLYDEKQQHIVYCSNDLLGDVFGVPSFSVKEHRKIYAMIYRNLVVVSQQDSGTSPS (SEQ ID NO: 34), and is selected from the group consisting of these.

[0123] In one example, the compositions described herein have an amino acid sequence having at least 90% sequence identity to the amino acid sequence of MAVPMQLSCSRQNGTWPREVLLVLSVNSSVFLHLQALGIPLHLAYNSSLVTFQEPPGVNTTELRSTGGVPVQGSKYAADRNHYRRYPRRRGPPRNYQQNTRGLNAHGAAQMQPMHRYDVSALQYNSMTSSQTYMNGSPTYSMSYSQQGTPGMALGSMGSVRSQLRSLKERNPLKIFPSKRILRRHKRDWVVAPISVPENGKGPFPQRLNQLKSNKDRDTKIFYSITGPGADSPPEGVFAVEKETRSAGETYTMKEVLFYLGQYIMTKRLYDEKQQHIVYCSNDLLGDLFGVPSFSVKEHRKIYTMIYRNLVVVNQQESSDSGTSVSRS (SEQ ID NO: 39); an amino acid sequence having at least 90% sequence identity to the amino acid sequence of MAVPMTVFMRLNIISPDLSGCTSKGLVLPAVLGITFGAFLIGALLTAALWYIYSHTRSPSKREPVVAVAAPASSESSSTNHSIGSTQSTPCSTSSMATGGVPVQGSKYAADRNHYRRYPRRRGPPRNYQQNTRGLNAHGAAQMQPMHRYDVSALQYNSMTSSQTYMNGSPTYSMSYSQQGTPGMALGSMGSVRSQLLKIFPSKRILRRHKRDWVVAPISVPENGKGPFPQRLNQLKSNKDRDTKIFYSITGPGADSPPEGVFAVEKETRSAGETYTMKEVLFYLGQYIMTKRLYDEKQQHIVYCSNDLLGDLFGVPSFSVKEHRKIYTMIYRNLVVVNQQESSDSGTSVSRS (SEQ ID NO: 40);An amino acid sequence having at least 90% sequence identity to the amino acid sequence of MAVPMTVSMRLNIVSPDLSGKGLVLPSVLGITFGAFLIGALLTAALWYIYSHTRGPSKREPVVAVAAPASSESSSTNHSIGSTQSTPCSTSSMATGGVPVQGSKYAADRNHYRRYPRRRGPPRNYQQNTRGLNAHGAAQMQPMHRYDVSALQYNSMTSSQTYMNGSPTYSMSYSQQGTPGMALGSMGSVRSQLAMHSPPTRILRRRKREWVMPPIFVPENGKGPFPQRLNQLKSNKDRGTKIFYSITGPGADSPPEGVFTIEKESRSAGETYTMKEIIFYIGQYIMTKRLYDEKQQHIVYCSNDLLGDVFGVPSFSVKEHRKIYAMIYRNLVAVSQQDSGTSLSRS (SEQ ID NO: 41); or an amino acid sequence having at least 90% sequence identity to the amino acid sequence of MAVPMTVSMRLNIVSPDLSGKGLVLPSVLGITFGAFLIGALLTAALWYIYSHTRAPSKREPVVAVAAPASSESSSTNHSIGSTQSTPCSTSSMATGGVPVQGSKYAADRNHYRRYPRRRGPPRNYQQNTRGLNAHGAAQMQPMHRYDVSALQYNSMTSSQTYMNGSPTYSMSYSQQGTPGMALGSMGSVRSQLVMNSPPSRILRRRKREWVMPPISVPENGKGPFPQRLNQLKSNKDRGTKLFYSITGPGADSPPEGVFTIEKETRSAGEIYTMKEIIFYIGQYIMTKRLYDEKQQHIVYCSNDLLGDVFGVPSFSVKEHRKIYAMIYRNLVVVSQQDSGTSPSRS (SEQ ID NO: 42), and comprises the amino acid sequence of a fusion peptide of 5 epitopes selected from the group consisting of such amino acid sequences.;

[0124] The composition described in this specification is a composition comprising a first plasmid containing a first nucleotide sequence encoding a first epitope of a first antigen, wherein the first epitope is a portion of a peptide selected from IGFBP-2, HER-2 or IGF-1R, and the first nucleotide sequence can comprise the composition located on the plasmid. In some examples, the composition can comprise a first plasmid containing a first nucleotide sequence encoding a first epitope of a first antigen and a second nucleotide sequence encoding a second epitope of a second antigen, wherein the first and second epitopes are independently selected from IGFBP-2, HER-2 or IGF-1R, and the first nucleotide sequence and the second nucleotide sequence are located on one or more plasmids. In some examples, at least the first epitope of the peptide IGFBP-2 has an amino acid sequence having at least 90% sequence identity to the amino acid sequence of NHVDSTMNMLGGGGS (SEQ ID NO: 46); an amino acid sequence having at least 90% sequence identity to the amino acid sequence of ELAVFREKVTEQHRQ (SEQ ID NO: 47); an amino acid sequence having at least 90% sequence identity to the amino acid sequence of LGLEEPKKLRPPPAR (SEQ ID NO: 48); an amino acid sequence having at least 90% sequence identity to the amino acid sequence of DQVLERISTMRLPDE (SEQ ID NO: 49); an amino acid sequence having at least 90% sequence identity to the amino acid sequence of GPLEHLYSLHIPNCD (SEQ ID NO: 50); an amino acid sequence having at least 90% sequence identity to the amino acid sequence of KHGLYNLKQCKMSLN (SEQ ID NO: 51); an amino acid sequence having at least 90% sequence identity to the amino acid sequence of PECHLFYNEQQEARG (SEQ ID NO: 53); an amino acid sequence having at least 90% sequence identity to the amino acid sequence of MLPRVGCPALPLPPPPLLPLLPLLLLLLGASGGGGGARAEVLFRCPPCTPERLAACGPPPVAPPAAVAAVAGGARMPCAELVREPGCGCCSVCARLEGEACGVYTPRCGQGLRCYPHPGSELPLQALVMGEGTCEKRRDAEYGASPEQVADNGDDHSEGGLVE (SEQ ID NO: 54);An amino acid sequence having at least 90% sequence identity to the amino acid sequence of MLPRLGGPALPLLLPSLLLLLLLGAGGCGPGVRAEVLFRCPPCTPERLAACGPPPDAPCAELVREPGCGCCSVCARQEGEACGVYIPRCAQTLRCYPNPGSELPLKALVTGAGTCEKRRVGTTPQQVADSDDDHSEGGLVE (SEQ ID NO: 55); or selected from the group consisting of amino acid sequences having at least 90% sequence identity to the amino acid sequence of MLPRLGGPALPLLLPSLLLLLLLGAGGCGPGVRAEVLFRCPPCTPERLAACGPPPDAPCAELVREPGCGCCSVCARQEGEACGVYIPRCAQTLRCYPNPGSELPLKALVTGAGTCEKRRVGATPQQVADSEDDHSEGGLVE (SEQ ID NO: 56). In some examples, at least the first epitope of the peptide HER-2 is relative to the amino acid sequence of TMRRLLQETELVEPLTPSGAMPNQAQMRILKETELRKVKVLGSGAFGTVYKGIWIPDGENVKIPVAIKVLRENTSPKANKEILDEAYVMAGVGSPYVSRLLGICLTSTVQLVTQLMPYGCLLDHVRENRGRLGSQDLLNWCMQIAKGMSYLEDVRLVHRDLAARNVLVKSPNHVKITDFGLARLLDIDETEYHADGGKVPIKWMALESILRRRFTHQSDVWSYGVTVWELMTFGAKPYDGIPAREIPDLLEKGERLPQPPICTIDVYMIMVKCWMIDSECRPRFRELVSEFSRMARDPQRFVVIQNEDLAPGAGGMVHHRHRSSSPLPAARPAGATLERPKTLSPGKNGVVKDVFAFGGAVENPEYL (SEQ ID NO: 60); A nucleotide sequence encoding an amino acid sequence having at least 90% sequence identity; relative to the amino acid sequence of TMRRLLQETELVEPLTPSGAVPNQAQMRILKETELRKLKVLGSGAFGTVYKGIWIPDGENVKIPVAIKVLRENTSPKANKEILDEAYVMAGVGSPYVSRLLGICLTSTVQLVTQLMPYGCLLDHVREHRGRLGSQDLLNWCVQIAKGMSYLEEVRLVHRDLAARNVLVKSPNHVKITDFGLARLLDIDETEYHADGGKVPIKWMALESILRRRFTHQSDVWSYGVTVWELMTFGAKPYDGIPAREIPDLLEKGERLPQPPICTIDVYMIMVKCWMIDSECRPRFRELVSEFSRMARDPQRFVVIQNEDLALGTGSTAHRRHRSSSPPPPIRPAGATLERPKTLSPGKNGVVKDVFAFGGAVENPEYL (SEQ ID NO: 61) A nucleotide sequence encoding an amino acid sequence having at least 90% sequence identity; or the amino acid sequence of TMRRLLQETELVEPLTPSGAMPNQAQMRILKETELRKVKVLGSGAFGTVYKGIWIPDGENVKIPVAIKVLRENTSPKANKEILDEAYVMAGVGSPYVSRLLGICLTSTVQLVTQLMPYGCLLDHVREHRGRLGSQDLLNWCVQIAKGMSYLEDVRLVHRDLAARNVLVKSPNHVKITDFGLARLLDIDETEYHADGGKVPIKWMALESILRRRFTHQSDVWSYGVTVWELMTFGAKPYDGIPAREIPDLLEKGERLPQPPICTIDVYMIMVKCWMIDSECRPRFRELVSEFSRMARDPQRFVVIQNEDLTPGTGSTAHRRHRSSSPLPPVRPAGATLERPKTLSPGKNGVVKDVFAFGGAVENPEYL (SEQ ID NO: 62) It is selected from the group consisting of nucleotide sequences encoding amino acid sequences having at least 90% sequence identity thereto. In some examples, the nucleic acid sequence encoding an epitope of the peptide IGF-1R is an amino acid sequence having at least 90% sequence identity to the amino acid sequence of DYRSYRFPKLTVITE (SEQ ID NO: 66); an amino acid sequence having at least 90% sequence identity to the amino acid sequence of IRGWKLFYNYALVIF (SEQ ID NO: 67); an amino acid sequence having at least 90% sequence identity to the amino acid sequence of VVTGYVKIRHSHALV (SEQ ID NO: 68); an amino acid sequence having at least 90% sequence identity to the amino acid sequence of FFYVQAKTGYENFIH (SEQ ID NO: 69); an amino acid sequence having at least 90% sequence identity to the amino acid sequence of LIIALPVAVLLIVGG (SEQ ID NO: 70); an amino acid sequence having at least 90% sequence identity to the amino acid sequence of LVIMLYVFHRKRNNS (SEQ ID NO: 71); an amino acid sequence having at least 90% sequence identity to the amino acid sequence of NCHHVVRLLGVVSQG (SEQ ID NO: 72); an amino acid sequence having at least 90% sequence identity to the amino acid sequence of WSFGVVLWEIATLAEQPYQGLSNEQVLRFVMEGGLLDKPDNCPDMLFELMRMCWQYNPKMRPSFLEHKAENGPGPGVLVLRASFDERQPYAHMNGGRKNERALP (SEQ ID NO: 73); an amino acid sequence having at least 90% sequence identity to the amino acid sequence of WSFGVVLWEIATLAEQPYQGLSNEQVLRFVMEGGLLDKPDNCPDMLFELMRMCWQYNPKMRPSFLEHKAENGPGPGVLVLRASFDERQPYAHMNGGRANERALP (SEQ ID NO: 74); or an amino acid sequence having at least 90% sequence identity to the amino acid sequence of WSFGVVLWEIATLAEQPYQGLSNEQVLRFVMEGGLLDKPDNCPDMLFELMRMCWQYNPKMRPSFLEHKAENGPGVLVLRASFDERQPYAHMNGGRANERALP (SEQ ID NO: 75), and is selected from the group consisting thereof.

[0125] The composition described in this specification has an amino acid sequence having at least 90% sequence identity to the amino acid sequence of MAVPMLPRVGCPALPLPPPPLLPLLPLLLLLLGASGGGGGARAEVLFRCPPCTPERLAACGPPPVAPPAAVAAVAGGARMPCAELVREPGCGCCSVCARLEGEACGVYTPRCGQGLRCYPHPGSELPLQALVMGEGTCEKRRDAEYGASPEQVADNGDDHSEGGLVEQLTMRRLLQETELVEPLTPSGAMPNQAQMRILKETELRKVKVLGSGAFGTVYKGIWIPDGENVKIPVAIKVLRENTSPKANKEILDEAYVMAGVGSPYVSRLLGICLTSTVQLVTQLMPYGCLLDHVRENRGRLGSQDLLNWCMQIAKGMSYLEDVRLVHRDLAARNVLVKSPNHVKITDFGLARLLDIDETEYHADGGKVPIKWMALESILRRRFTHQSDVWSYGVTVWELMTFGAKPYDGIPAREIPDLLEKGERLPQPPICTIDVYMIMVKCWMIDSECRPRFRELVSEFSRMARDPQRFVVIQNEDLAPGAGGMVHHRHRSSSPLPAARPAGATLERPKTLSPGKNGVVKDVFAFGGAVENPEYLGRPVPWSFGVVLWEIATLAEQPYQGLSNEQVLRFVMEGGLLDKPDNCPDMLFELMRMCWQYNPKMRPSFLEHKAENGPGPGVLVLRASFDERQPYAHMNGGRKNERALPAAA (SEQ ID NO: 79);An amino acid sequence having at least 90% sequence identity to the amino acid sequence of MAVPMLPRLGGPALPLLLPSLLLLLLLGAGGCGPGVRAEVLFRCPPCTPERLAACGPPPDAPCAELVREPGCGCCSVCARQEGEACGVYIPRCAQTLRCYPNPGSELPLKALVTGAGTCEKRRVGTTPQQVADSDDDHSEGGLVEQLTMRRLLQETELVEPLTPSGAVPNQAQMRILKETELRKLKVLGSGAFGTVYKGIWIPDGENVKIPVAIKVLRENTSPKANKEILDEAYVMAGVGSPYVSRLLGICLTSTVQLVTQLMPYGCLLDHVREHRGRLGSQDLLNWCVQIAKGMSYLEEVRLVHRDLAARNVLVKSPNHVKITDFGLARLLDIDETEYHADGGKVPIKWMALESILRRRFTHQSDVWSYGVTVWELMTFGAKPYDGIPAREIPDLLEKGERLPQPPICTIDVYMIMVKCWMIDSECRPRFRELVSEFSRMARDPQRFVVIQNEDLALGTGSTAHRRHRSSSPPPPIRPAGATLERPKTLSPGKNGVVKDVFAFGGAVENPEYLGRPVPWSFGVVLWEIATLAEQPYQGLSNEQVLRFVMEGGLLDKPDNCPDMLFELMRMCWQYNPKMRPSFLEHKAENGPGPGVLVLRASFDERQPYAHMNGGRANERALPAAA (SEQ ID NO: 80);Or it can further contain a nucleic acid sequence encoding a fusion protein of three epitopes selected from the group consisting of amino acid sequences having at least 90% sequence identity to the amino acid sequence of MAVPMLPRLGGPALPLLLPSLLLLLLLGAGGCGPGVRAEVLFRCPPCTPERLAACGPPPDAPCAELVREPGCGCCSVCARQEGEACGVYIPRCAQTLRCYPNPGSELPLKALVTGAGTCEKRRVGATPQQVADSEDDHSEGGLVEQLTMRRLLQETELVEPLTPSGAMPNQAQMRILKETELRKVKVLGSGAFGTVYKGIWIPDGENVKIPVAIKVLRENTSPKANKEILDEAYVMAGVGSPYVSRLLGICLTSTVQLVTQLMPYGCLLDHVREHRGRLGSQDLLNWCVQIAKGMSYLEDVRLVHRDLAARNVLVKSPNHVKITDFGLARLLDIDETEYHADGGKVPIKWMALESILRRRFTHQSDVWSYGVTVWELMTFGAKPYDGIPAREIPDLLEKGERLPQPPICTIDVYMIMVKCWMIDSECRPRFRELVSEFSRMARDPQRFVVIQNEDLTPGTGSTAHRRHRSSSPLPPVRPAGATLERPKTLSPGKNGVVKDVFAFGGAVENPEYLGRPVPWSFGVVLWEIATLAEQPYQGLSNEQVLRFVMEGGLLDKPDNCPDMLFELMRMCWQYNPKMRPSFLEHKAENGPGVLVLRASFDERQPYAHMNGGRANERALPAAA (SEQ ID NO: 81).;

[0126] In some examples, the composition comprises first and second epitopes independently selected from CD105, Yb-1, SOX-2, CDH3, or MDM2. In some examples, the composition further comprises a third epitope, and the first, second, and third epitopes are independently selected from CD105, Yb-1, SOX-2, CDH3, or MDM2. In some examples, the composition further comprises third and fourth epitopes, and the first, second, third, and fourth epitopes are independently selected from CD105, Yb-1, SOX-2, CDH3, or MDM2. In some examples, the composition further comprises third, fourth, and fifth epitopes, and the first, second, third, fourth, and fifth epitopes are independently selected from CD105, Yb-1, SOX-2, CDH3, or MDM2.

[0127] In some examples, the composition comprises first and second epitopes independently selected from IGFBP2, HER-2, or IGF-1R. In some examples, the composition further comprises a third epitope, and the first, second, and third epitopes are independently selected from IGFBP2, HER-2, or IGF-1R.

[0128] In some examples, the composition can comprise at least a first epitope of a first antigen, and the first epitope is a portion of a peptide derived from HIF-1α. In some examples, the composition can comprise at least a first epitope of a first antigen and at least a second epitope of a second antigen, and the first and second epitopes are derived from HIF-1α.

[0129] In some examples, the composition can include a nucleic acid sequence encoding an epitope of peptide HIF-1α selected from the group consisting of a nucleotide sequence encoding an amino acid sequence having at least 90% sequence identity to the amino acid sequence of DSKTFLSRHSLDMKFSYCDERITELMGYEPEELLGRSIYEYYHALDSDHLTKTHHDMFTKGQVTTGQYRMLAKRGGYVWVETQATVIYN (SEQ ID NO: 82); a nucleotide sequence encoding an amino acid sequence having at least 90% sequence identity to the amino acid sequence of SDNVNKYMGLTQFELTGHSVFDFTHP (SEQ ID NO: 83); and a nucleotide sequence encoding an amino acid sequence having at least 90% sequence identity to the amino acid sequence of GGYVWVETQATVIYNTKNSQ (SEQ ID NO: 84).

[0130] In some examples, the composition can include at least a first epitope of peptide HIF-1α selected from the group consisting of an amino acid sequence having at least 90% sequence identity to the amino acid sequence of DSKTFLSRHSLDMKFSYCDERITELMGYEPEELLGRSIYEYYHALDSDHLTKTHHDMFTKGQVTTGQYRMLAKRGGYVWVETQATVIYN (SEQ ID NO: 82); an amino acid sequence having at least 90% sequence identity to the amino acid sequence of SDNVNKYMGLTQFELTGHSVFDFTHP (SEQ ID NO: 83); and an amino acid sequence having at least 90% sequence identity to the amino acid sequence of GGYVWVETQATVIYNTKNSQ (SEQ ID NO: 84).

[0131] The amino acid sequences of epitopes from the following proteins, CD105, HIF1α, MDM2, Yb1, SOX-2, HER-2, IGFBP2, IGF-1R and CDH3, can be different from those provided herein. In some examples, amino acid sequences that are 95%, 90%, 85%, 80%, 75%, 70%, 65%, 60%, greater than 55% or greater than 50% homologous to the amino acid sequences disclosed herein can be used in the compositions described herein.

[0132] In some examples, the first amino acid sequence is selected from the group of species consisting of human, mouse, and rat. In some examples, the second amino acid sequence is selected from the group of species consisting of human, mouse, and rat.

[0133] In some examples, the first and second nucleic acid sequences are located on the first plasmid. In some examples, the second nucleic acid sequence is located on the second plasmid. In some examples, the amino acid sequences of the first and second epitopes are separated by a linker amino acid sequence. In some examples, the amino acid sequence of the first epitope is adjacent to the amino acid sequence of the second epitope.

[0134] In some examples, the immune response is a type 1 immune response. In some examples, the immune response is characterized by the ratio of type I cytokine production to type II cytokine production greater than 1. In some examples, the immune response is characterized by the ratio of type I cytokine production to type II cytokine production less than 1. In some examples, the immune response is characterized by the ratio of IFNγ production to IL-10 production greater than 1. In some examples, the immune response is characterized by the ratio of IFNγ production to IL-10 production less than 1.

[0135] In some examples, the composition is administered to a subject. In some examples, the subject is in need of administration of the composition. In some examples, the composition is effective in inducing an immune response in a subject. In some examples, the composition is effective in eliminating a number of cells associated with breast cancer in a subject. In some examples, the composition can be used to prevent the growth of cells associated with breast cancer in a subject.

[0136] In some examples, the subject is selected from the group consisting of a human having breast cancer, a mouse having breast cancer, and a rat having breast cancer. In some examples, the subject is selected from the group consisting of a human without breast cancer, a mouse without breast cancer, and a rat without breast cancer.

[0137] In some examples, the cells associated with breast cancer are selected from breast cells expressing atypia features, pre-tumor breast cells, breast cancer cells, pre-invasive breast cancer cells, breast cancer stem cells, epithelial cells, mesenchymal cells, stromal cells, or combinations thereof.

[0138] In some examples, at least the first epitope is contained within a pharmaceutical composition. In some examples, at least the first epitope is contained within a pharmaceutical composition further comprising a pharmaceutical carrier. In some examples, at least the first epitope is contained within a pharmaceutical composition further comprising a pharmaceutical carrier and an adjuvant. In some examples, at least the first epitope is contained within a pharmaceutical composition further comprising an adjuvant. In some examples, the composition further comprises an adjuvant and a pharmaceutical carrier. In some examples, the adjuvant is GM-CSF.

[0139] In some examples, the composition can be administered to a subject. In some examples, the subject is in need thereof. In some examples, a method for preventing breast cancer in a subject is provided herein such that the method comprises the step of administering the composition described herein to the subject. In some examples, a method for treating breast cancer in a subject is provided herein such that the method comprises the step of administering the composition described herein to the subject. In some examples, the step of administering further comprises delivering at least one dose of the composition described herein to the subject. In some examples, the step of administering further comprises delivering the composition described herein to the subject by subcutaneous injection, intradermal injection, intramuscular injection, intravascular injection, topical application, or inhalation. In some examples, the subject is selected from the group consisting of a human having breast cancer, a mouse having breast cancer, and a rat having breast cancer. In some examples, the subject is selected from the group consisting of a human without breast cancer, a mouse without breast cancer, and a rat without breast cancer.

[0140] The present disclosure also provides a kit for preparing the compositions described herein, the kit including instructions for preparing the compositions. The present disclosure also provides a kit for administering the compositions described herein, the kit including instructions for administering the compositions. A composition comprising an epitope selected from survivin, HIF-1α, IGFBP-2, and IGF-1R

[0141] The compositions described herein include a plasmid comprising at least one nucleotide sequence encoding a polypeptide comprising at least 70% sequence identity to an epitope sequence selected from SEQ ID NOs: 54, 73, 85, and 87, and an excipient. The composition can include a plasmid comprising at least one nucleotide sequence encoding a polypeptide comprising at least 80% sequence identity to an epitope sequence selected from SEQ ID NOs: 54, 73, 85, and 87. The composition can include a plasmid comprising at least one nucleotide sequence encoding a polypeptide comprising at least 90% sequence identity to an epitope sequence selected from SEQ ID NOs: 54, 73, 85, and 87. The composition can include a plasmid comprising at least one nucleotide sequence encoding a polypeptide comprising at least 95% sequence identity to an epitope sequence selected from SEQ ID NOs: 54, 73, 85, and 87. The composition can include a plasmid comprising at least one nucleotide sequence encoding a polypeptide comprising at least 99% sequence identity to an epitope sequence selected from SEQ ID NOs: 54, 73, 85, and 87. The composition can include a plasmid comprising at least one nucleotide sequence encoding a polypeptide comprising 100% sequence identity to an epitope sequence selected from SEQ ID NOs: 54, 73, 85, and 87. The composition can include a plasmid comprising at least one nucleotide sequence encoding a polypeptide consisting of 100% sequence identity to the full length of an epitope sequence selected from SEQ ID NOs: 54, 73, 85, and 87.

[0142] Optionally, at least one nucleotide sequence can encode a polypeptide that contains at least 70% sequence identity to at least 20 contiguous amino acids of SEQ ID NO: 85. At least one nucleotide sequence can encode a polypeptide that contains at least 80% sequence identity to at least 20 contiguous amino acids of SEQ ID NO: 85. At least one nucleotide sequence can encode a polypeptide that contains at least 90% sequence identity to at least 20 contiguous amino acids of SEQ ID NO: 85. At least one nucleotide sequence can encode a polypeptide that contains at least 95% sequence identity to at least 20 contiguous amino acids of SEQ ID NO: 85. At least one nucleotide sequence can encode a polypeptide that contains at least 99% sequence identity to at least 20 contiguous amino acids of SEQ ID NO: 85. At least one nucleotide sequence can encode a polypeptide that contains at least 100% sequence identity to at least 20 contiguous amino acids of SEQ ID NO: 85.

[0143] Optionally, at least one nucleotide sequence can encode a polypeptide that contains at least 70% sequence identity to at least 60 contiguous amino acids of SEQ ID NO: 87. At least one nucleotide sequence can encode a polypeptide that contains at least 80% sequence identity to at least 60 contiguous amino acids of SEQ ID NO: 87. At least one nucleotide sequence can encode a polypeptide that contains at least 90% sequence identity to at least 60 contiguous amino acids of SEQ ID NO: 87. At least one nucleotide sequence can encode a polypeptide that contains at least 95% sequence identity to at least 60 contiguous amino acids of SEQ ID NO: 87. At least one nucleotide sequence can encode a polypeptide that contains at least 99% sequence identity to at least 60 contiguous amino acids of SEQ ID NO: 87. At least one nucleotide sequence can encode a polypeptide that contains at least 100% sequence identity to at least 60 contiguous amino acids of SEQ ID NO: 87.

[0144] In some cases, the composition comprises an isolated plasmid comprising at least four nucleotide sequences. Optionally, each of the at least four nucleotide sequences independently encodes a polypeptide comprising at least 60%, 70%, 80%, 90%, 95%, 99% or 100% sequence identity to an epitope sequence selected from SEQ ID NO: 54, 73, 85 and 87. Optionally, the isolated plasmid can comprise four nucleotide sequences. Optionally, each of the four nucleotide sequences can independently encode a polypeptide comprising at least 60%, 70%, 80%, 90%, 95%, 99% or 100% sequence identity to an epitope sequence selected from SEQ ID NO: 54, 73, 85 and 87. In other cases, each of the four nucleotide sequences can encode a different polypeptide. Optionally, each of the different polypeptides can comprise at least 60%, 70%, 80%, 90%, 95%, 99% or 100% sequence identity to an epitope sequence selected from SEQ ID NO: 54, 73, 85 and 87.

[0145] Optionally, one of the four nucleotide sequences can encode a polypeptide comprising at least 70% sequence identity to at least 20 contiguous amino acids of SEQ ID NO: 85. One of the four nucleotide sequences can encode a polypeptide comprising at least 80% sequence identity to at least 20 contiguous amino acids of SEQ ID NO: 85. One of the four nucleotide sequences can encode a polypeptide comprising at least 90% sequence identity to at least 20 contiguous amino acids of SEQ ID NO: 85. One of the four nucleotide sequences can encode a polypeptide comprising at least 95% sequence identity to at least 20 contiguous amino acids of SEQ ID NO: 85. One of the four nucleotide sequences can encode a polypeptide comprising at least 100% sequence identity to at least 20 contiguous amino acids of SEQ ID NO: 85.

[0146] Optionally, one of the four nucleotide sequences can encode a polypeptide that contains at least 70% sequence identity to at least 60 consecutive amino acids of SEQ ID NO: 87. One of the four nucleotide sequences can encode a polypeptide that contains at least 80% sequence identity to at least 60 consecutive amino acids of SEQ ID NO: 87. One of the four nucleotide sequences can encode a polypeptide that contains at least 90% sequence identity to at least 60 consecutive amino acids of SEQ ID NO: 87. One of the four nucleotide sequences can encode a polypeptide that contains at least 95% sequence identity to at least 60 consecutive amino acids of SEQ ID NO: 87. One of the four nucleotide sequences can encode a polypeptide that contains at least 100% sequence identity to at least 60 consecutive amino acids of SEQ ID NO: 87.

[0147] In some examples, the four nucleotide sequences are arranged tandemly within a plasmid. The four nucleotide sequences can be separated by the sequence of a linker nucleic acid. The sequence of the linker nucleic acid can be about 1 to about 150, about 5 to about 100, or about 10 to about 50 nucleic acids in length. In some cases, the nucleic acid can encode one or more amino acid residues. Optionally, the amino acid sequence of the linker can be about 1 to about 50 or about 5 to about 25 amino acid residues in length. Optionally, the linker can include the linker shown as the underlined portion in FIG. 14 (SEQ ID NO: 14).

[0148] Optionally, the composition can further include at least one additional isolated plasmid. Optionally, the composition can further include at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 30, 35, 40, 45, 50 or more additional isolated plasmids.

[0149] In some examples, at least one additional isolated plasmid comprises a nucleotide sequence encoding a polypeptide comprising at least 70% sequence identity to an epitope sequence selected from SEQ ID NOs: 54, 73, 85, and 87. At least one additional isolated plasmid can comprise a nucleotide sequence encoding a polypeptide comprising at least 80% sequence identity to an epitope sequence selected from SEQ ID NOs: 54, 73, 85, and 87. At least one additional isolated plasmid can comprise a nucleotide sequence encoding a polypeptide comprising at least 90% sequence identity to an epitope sequence selected from SEQ ID NOs: 54, 73, 85, and 87. At least one additional isolated plasmid can comprise a nucleotide sequence encoding a polypeptide comprising at least 95% sequence identity to an epitope sequence selected from SEQ ID NOs: 54, 73, 85, and 87. At least one additional isolated plasmid can comprise a nucleotide sequence encoding a polypeptide comprising at least 99% sequence identity to an epitope sequence selected from SEQ ID NOs: 54, 73, 85, and 87. At least one additional isolated plasmid can comprise a nucleotide sequence encoding a polypeptide comprising 100% sequence identity to an epitope sequence selected from SEQ ID NOs: 54, 73, 85, and 87.

[0150] The composition described herein can include a plasmid containing a nucleotide sequence encoding a polypeptide having at least 80% sequence identity to SEQ ID NO: 89, and an excipient. The composition can include a plasmid containing a nucleotide sequence encoding a polypeptide having at least 90% sequence identity to SEQ ID NO: 89. The composition can include a plasmid containing a nucleotide sequence encoding a polypeptide having at least 95% sequence identity to SEQ ID NO: 89. The composition can include a plasmid containing a nucleotide sequence encoding a polypeptide having 100% sequence identity to SEQ ID NO: 89. The composition can include a plasmid containing a nucleotide sequence encoding a polypeptide consisting of 100% sequence identity to SEQ ID NO: 89.

[0151] The composition described herein can include a composition containing a polypeptide having at least 80% sequence identity to SEQ ID NO: 89. The composition can include a polypeptide having at least 90% sequence identity to SEQ ID NO: 89. The composition can include a polypeptide having at least 95% sequence identity to SEQ ID NO: 89. The composition can include a polypeptide having 100% sequence identity to SEQ ID NO: 89. The composition can include a polypeptide consisting of 100% sequence identity to SEQ ID NO: 89.

[0152] The composition can be formulated for the treatment of breast cancer or ovarian cancer in a subject. The breast cancer can be recurrent or refractory breast cancer. The ovarian cancer can be recurrent or refractory ovarian cancer. The breast cancer can be metastatic breast cancer. The ovarian cancer can be metastatic ovarian cancer.

[0153] The composition can induce an immune response. The immune response can be characterized by the ratio of type I cytokine production to type II cytokine production that exceeds 1. The immune response can be characterized by the ratio of type I cytokine production to type II cytokine production that is less than 1. The immune response can be characterized by the ratio of IFNγ production to IL-10 production that exceeds 1. The immune response can be characterized by the ratio of IFNγ production to IL-10 production that is less than 1.

[0154] Optionally, the composition can further comprise an adjuvant. Optionally, the adjuvant is GM-CSF.

[0155] The present composition can further comprise an excipient. The excipient can be a pharmaceutically acceptable carrier.

[0156] Often, the composition can be formulated for subcutaneous, intramuscular or intradermal administration.

[0157] The present disclosure also provides a kit for preparing the composition described herein, the kit comprising instructions for preparing the composition. The present disclosure also provides a kit for administering the composition described herein, the kit comprising instructions for administering the composition. Plasmid for pharmaceutical composition

[0158] In some examples, the epitope can be derived from a human protein that can be used directly in a peptide-based vaccine. In other examples, the epitope can be derived from a human protein, and the nucleic acid sequence encoding the epitope can be incorporated into a nucleic acid construct designed to induce the expression of the epitope in a subject after administration. For example, the encoded epitope from the nucleic acid construct can enable an immune response against at least one epitope to be coordinated, amplified, attenuated, suppressed, or eliminated with respect to a specific set of proteins (e.g., self-proteins). In some examples, the peptide or nucleic acid construct can be optimized for protein- or plasmid-based vaccination to induce, amplify, or coordinate a Th1 immune response. In some examples, the epitope can be an extended Th1 epitope. In other examples, the peptide or nucleic acid construct can be optimized for protein- or plasmid-based vaccination to suppress, attenuate, or eliminate a pathological response in a subject (e.g., a human or an animal) that requires it.

[0159] The compositions described herein can include a plasmid containing a nucleic acid sequence for expressing at least one epitope in a subject after administration of the composition (e.g., a vaccine). Known to those of ordinary skill in the art suitable for pharmaceutical use for the expression of the nucleic acid sequence Any plasmid backbone (e.g., a vector) can be used in the compositions described herein. In some examples, a commercially available plasmid backbone can be used. For example, plasmid pUMVC3 can be used. In some examples, a commercially available plasmid backbone can be modified, mutated, genetically engineered, or cloned before use. In other examples, a non-commercially available plasmid backbone can be used.

[0160] Before inserting the nucleic acid sequence of at least one epitope, the plasmid backbone can be less than about 500 bp, about 1.0 kB, about 1.2 kB, about 1.4 kB, about 1.6 kB, about 1.8 kB, about 2.0 kB, about 2.2 kB, about 2.4 kB, about 2.6 kB, about 2.8 kB, about 3.0 kB, about 3.2 kB, about 3.4 kB, about 3.6 kB, about 3.8 kB, about 4.0 kB, about 4.2 kB, about 4.4 kB, about 4.6 kB, about 4.8 kB, about 5.0 kB, about 5.2 kB, about 5.4 kB, about 5.6 kB, about 5.8 kB, about 6.0 kB, about 6.2 kB, about 6.4 kB, about 6.6 kB, about 6.8 kB, about 7.0 kB, about 7.2 kB, about 7.4 kB, about 7.6 kB, about 7.8 kB, about 8.0 kB, about 8.2 kB, about 8.4 kB, about 8.6 kB, about 8.8 kB, about 9.0 kB, about 9.2 kB, about 9.4 kB, about 9.6 kB, about 9.8 kB, about 10.0 kB, about 10.2 kB, about 10.4 kB, about 10.6 kB, about 10.8 kB, about 11.0 kB, about 11.2 kB, about 11.4 kB, about 11.6 kB, about 11.8 kB, about 12.0 kB, about 12.2 kB, about 12.4 kB, about 12.6 kB, about 12.8 kB, about 13.0 kB, about 13.2 kB, about 13.4 kB, about 13.6 kB, about 13.8 kB, about 14 kB, about 14.5 kB, about 15 kB, about 15.5 kB, about 16 kB, about 16.5 kB, about 17 kB, about 17.5 kB, about 18 kB, about 18.5 kB, about 19 kB, about 19.5 kB, about 20 kB, about 30 kB, about 40 kB, about 50 kB, about 60 kB, about 70 kB, about 80 kB, about 90 kB, about 100 kB, about 110 kB, about 120 kB, about 130 kB, about 140 kB, about 150 kB, about 160 kB, about 170 kB, about 180 kB, about 190 kB or about 200 kB in length. In an exemplary example, before the addition of the nucleic acid sequence encoding at least one epitope, the plasmid is about 4 kB in length.

[0161] In some examples, the compositions described herein can include one plasmid. In other examples, the compositions described herein can include more than one plasmid. For example, the compositions described herein can include two plasmids, three plasmids, four plasmids, five plasmids, six plasmids, seven plasmids, eight plasmids, nine plasmids, ten plasmids, eleven plasmids, twelve plasmids, thirteen plasmids, fourteen plasmids, fifteen plasmids, sixteen plasmids, seventeen plasmids, eighteen plasmids, nineteen plasmids, twenty plasmids, or more than twenty plasmids.

[0162] In some examples, the nucleic acid encoding at least one epitope of the plasmid can be deoxyribonucleic acid. For example, the deoxyribonucleic acid can be single-stranded, double-stranded, or complementary. The deoxyribonucleic acid can be derived from genomic, mitochondrial, or plasmid deoxyribonucleic acid. In other examples, the nucleic acid of the plasmid can be ribonucleic acid. For example, the ribonucleic acid can be single-stranded or double-stranded. In some examples, the ribonucleic acid can be micro, antisense, small hairpin type, small interfering, messenger, transfer, ribosomal, or others. In some examples, the nucleic acid of the plasmid can be a portion of deoxyribonucleic acid and a portion of ribonucleic acid.

[0163] The nucleic acid encoding at least one epitope of the plasmid can be derived from any species such that the epitope expressed from the nucleic acid elicits an immune response in a subject. In some examples, the subject can be a rodent, non-human primate, or human. The nucleic acid encoding the epitope of the plasmid can be isolated from any nucleic acid source using methods and techniques known to those of skill in the art. The nucleic acid encoding the epitope of the plasmid can be cloned into the plasmid backbone using methods and techniques known to those of skill in the art.

[0164] In some examples, the nucleic acid sequence encoding the epitope can be a nucleic acid sequence endogenous to the subject. For example, the nucleic acid sequence of CD105 from humans can be used to express CD105 in humans. In other examples, the nucleic acid sequence of the antigenic epitope can be a nucleic acid sequence exogenous to the subject. For example, the nucleic acid sequence of CD105 from non-humans can be used to express CD105 in humans.

[0165] In some examples, the nucleic acid sequence for expressing the antigenic epitope can be a wild-type nucleic acid sequence. For example, the naturally occurring nucleic acid sequence of CD105 in a certain genome can be used to express CD105 in a subject. In other examples, the nucleic acid sequence encoding the epitope can be a synthetic nucleic acid sequence. For example, the naturally occurring nucleic acid sequence of CD105 in a certain genome can be modified using molecular techniques known to those skilled in the art and used for the expression of CD105 in a subject.

[0166] In some examples, the nucleic acid sequence encoding the epitope can be a nucleic acid sequence endogenous to the subject. For example, the nucleic acid sequence of CD105 from humans can be used to express CD105 in humans. In other examples, the nucleic acid sequence of the antigenic epitope can be a nucleic acid sequence exogenous to the subject. For example, the nucleic acid sequence of CD105 from non-humans can be used to express CD105 in humans.

[0167] In some examples, the nucleic acid sequence for expressing the antigenic epitope can be a wild-type nucleic acid sequence. For example, the naturally occurring nucleic acid sequence of CD105 in a certain genome can be used to express CD105 in a subject. In other examples, the nucleic acid sequence encoding the epitope can be a synthetic nucleic acid sequence. For example, the naturally occurring nucleic acid sequence of CD105 in a certain genome can be modified using molecular techniques known to those skilled in the art and used for the expression of CD105 in a subject.

[0168] In some examples, the nucleic acid sequence encoding the epitope can be a nucleic acid sequence endogenous to the subject. For example, the nucleic acid sequence of human-derived HIF-1A can be used to express HIF-1A in humans. In other examples, the nucleic acid sequence of the antigenic epitope can be a nucleic acid sequence exogenous to the subject. For example, the nucleic acid sequence of non-human-derived HIF-1A can be used to express HIF-1A in humans.

[0169] In some examples, the nucleic acid sequence for expressing the antigenic epitope can be a wild-type nucleic acid sequence. For example, the naturally-occurring nucleic acid sequence of HIF-1A in a certain genome can be used to express HIF-1A in a subject. In other examples, the nucleic acid sequence encoding the epitope can be a synthetic nucleic acid sequence. For example, the naturally-occurring nucleic acid sequence of HIF-1A in a certain genome can be modified using molecular techniques known to those skilled in the art and used for the expression of HIF-1A in a subject.

[0170] In some examples, the nucleic acid sequence encoding the epitope can be a nucleic acid sequence endogenous to the subject. For example, the nucleic acid sequence of human-derived HIF-1A can be used to express HIF-1A in humans. In other examples, the nucleic acid sequence of the antigenic epitope can be a nucleic acid sequence exogenous to the subject. For example, the nucleic acid sequence of non-human-derived HIF-1A can be used to express HIF-1A in humans.

[0171] In some examples, the nucleic acid sequence for expressing the antigenic epitope can be a wild-type nucleic acid sequence. For example, the naturally-occurring nucleic acid sequence of HIF-1A in a certain genome can be used to express HIF-1A in a subject. In other examples, the nucleic acid sequence encoding the epitope can be a synthetic nucleic acid sequence. For example, the naturally-occurring nucleic acid sequence of HIF-1A in a certain genome can be modified using molecular techniques known to those skilled in the art and used for the expression of HIF-1A in a subject.

[0172] In some examples, the nucleic acid sequence encoding the epitope can be a nucleic acid sequence endogenous to the subject. For example, the nucleic acid sequence of MDM2 from humans can be used to express MDM2 in humans. In other examples, the nucleic acid sequence of the antigenic epitope can be a nucleic acid sequence exogenous to the subject. For example, the nucleic acid sequence of MDM2 from non-humans can be used to express MDM2 in humans.

[0173] In some examples, the nucleic acid sequence for expressing the antigenic epitope can be a wild-type nucleic acid sequence. For example, the nucleic acid sequence of the natural origin of MDM2 in a certain genome can be used to express MDM2 in a subject. In other examples, the nucleic acid sequence encoding the epitope can be a synthetic nucleic acid sequence. For example, the nucleic acid sequence of the natural origin of MDM2 in a certain genome can be modified using molecular techniques known to those skilled in the art and used for the expression of MDM2 in a subject.

[0174] In some examples, the nucleic acid sequence encoding the epitope can be a nucleic acid sequence endogenous to the subject. For example, the nucleic acid sequence of MDM2 from humans can be used to express MDM2 in humans. In other examples, the nucleic acid sequence of the antigenic epitope can be a nucleic acid sequence exogenous to the subject. For example, the nucleic acid sequence of MDM2 from non-humans can be used to express MDM2 in humans.

[0175] In some examples, the nucleic acid sequence for expressing the antigenic epitope can be a wild-type nucleic acid sequence. For example, the nucleic acid sequence of the natural origin of MDM2 in a certain genome can be used to express MDM2 in a subject. In other examples, the nucleic acid sequence encoding the epitope can be a synthetic nucleic acid sequence. For example, the nucleic acid sequence of the natural origin of MDM2 in a certain genome can be modified using molecular techniques known to those skilled in the art and used for the expression of MDM-2 in a subject.

[0176] In some examples, the nucleic acid sequence encoding the epitope can be a nucleic acid sequence endogenous to the subject. For example, the nucleic acid sequence of Yb-1 derived from humans can be used to express Yb-1 in humans. In other examples, the nucleic acid sequence of the antigenic epitope can be a nucleic acid sequence exogenous to the subject. For example, the nucleic acid sequence of Yb-1 derived from non-humans can be used to express Yb-1 in humans.

[0177] In some examples, the nucleic acid sequence for expressing the antigenic epitope can be a wild-type nucleic acid sequence. For example, the nucleic acid sequence of the natural origin of Yb-1 in a certain genome can be used to express Yb-1 in the subject. In other examples, the nucleic acid sequence encoding the epitope can be a synthetic nucleic acid sequence. For example, the nucleic acid sequence of the natural origin of Yb-1 in a certain genome can be modified using molecular techniques known to those skilled in the art and can be used for the expression of Yb-1 in the subject.

[0178] In some examples, the nucleic acid sequence encoding the epitope can be a nucleic acid sequence endogenous to the subject. For example, the nucleic acid sequence of Yb-1 derived from humans can be used to express Yb-1 in humans. In other examples, the nucleic acid sequence of the antigenic epitope can be a nucleic acid sequence exogenous to the subject. For example, the nucleic acid sequence of Yb-1 derived from non-humans can be used to express Yb-1 in humans.

[0179] In some examples, the nucleic acid sequence for expressing the antigenic epitope can be a wild-type nucleic acid sequence. For example, the nucleic acid sequence of the natural origin of Yb-1 in a certain genome can be used to express Yb-1 in the subject. In other examples, the nucleic acid sequence encoding the epitope can be a synthetic nucleic acid sequence. For example, the nucleic acid sequence of the natural origin of Yb-1 in a certain genome can be modified using molecular techniques known to those skilled in the art and can be used for the expression of Yb-1 in the subject.

[0180] In some examples, the nucleic acid sequence encoding the epitope can be a nucleic acid sequence endogenous to the subject. For example, the nucleic acid sequence of SOX-2 derived from humans can be used to express SOX-2 in humans. In other examples, the nucleic acid sequence of the antigenic epitope can be a nucleic acid sequence exogenous to the subject. For example, the nucleic acid sequence of SOX-2 derived from non-humans can be used to express SOX-2 in humans.

[0181] In some examples, the nucleic acid sequence for expressing the antigenic epitope can be a wild-type nucleic acid sequence. For example, the naturally occurring nucleic acid sequence of SOX-2 in a certain genome can be used to express SOX-2 in a subject. In other examples, the nucleic acid sequence encoding the epitope can be a synthetic nucleic acid sequence. For example, the naturally occurring nucleic acid sequence of SOX-2 in a certain genome can be modified using molecular techniques known to those skilled in the art and used for the expression of SOX-2 in a subject.

[0182] In some examples, the nucleic acid sequence encoding the epitope can be a nucleic acid sequence endogenous to the subject. For example, the nucleic acid sequence of SOX-2 derived from humans can be used to express SOX-2 in humans. In other examples, the nucleic acid sequence of the antigenic epitope can be a nucleic acid sequence exogenous to the subject. For example, the nucleic acid sequence of SOX-2 derived from non-humans can be used to express SOX-2 in humans.

[0183] In some examples, the nucleic acid sequence for expressing the antigenic epitope can be a wild-type nucleic acid sequence. For example, the naturally occurring nucleic acid sequence of SOX-2 in a certain genome can be used to express SOX-2 in a subject. In other examples, the nucleic acid sequence encoding the epitope can be a synthetic nucleic acid sequence. For example, the naturally occurring nucleic acid sequence of SOX-2 in a certain genome can be modified using molecular techniques known to those skilled in the art and used for the expression of SOX-2 in a subject.

[0184] In some examples, the nucleic acid sequence encoding the epitope can be a nucleic acid sequence endogenous to the subject. For example, the nucleic acid sequence of HER-2 derived from humans can be used to express HER-2 in humans. In other examples, the nucleic acid sequence of the antigenic epitope can be a nucleic acid sequence exogenous to the subject. For example, the nucleic acid sequence of HER-2 derived from non-humans can be used to express HER-2 in humans.

[0185] In some examples, the nucleic acid sequence for expressing the antigenic epitope can be a wild-type nucleic acid sequence. For example, the nucleic acid sequence of the natural origin of HER-2 in a certain genome can be used to express HER-2 in a subject. In other examples, the nucleic acid sequence encoding the epitope can be a synthetic nucleic acid sequence. For example, the nucleic acid sequence of the natural origin of HER-2 in a certain genome can be modified using molecular techniques known to those skilled in the art and can be used for the expression of HER-2 in a subject.

[0186] In some examples, the nucleic acid sequence encoding the epitope can be a nucleic acid sequence endogenous to the subject. For example, the nucleic acid sequence of HER-2 derived from humans can be used to express HER-2 in humans. In other examples, the nucleic acid sequence of the antigenic epitope can be a nucleic acid sequence exogenous to the subject. For example, the nucleic acid sequence of HER-2 derived from non-humans can be used to express HER-2 in humans.

[0187] In some examples, the nucleic acid sequence for expressing the antigenic epitope can be a wild-type nucleic acid sequence. For example, the nucleic acid sequence of the natural origin of HER-2 in a certain genome can be used to express HER-2 in a subject. In other examples, the nucleic acid sequence encoding the epitope can be a synthetic nucleic acid sequence. For example, the nucleic acid sequence of the natural origin of HER-2 in a certain genome can be modified using molecular techniques known to those skilled in the art and can be used for the expression of HER-2 in a subject.

[0188] In some examples, the nucleic acid sequence encoding the epitope can be a nucleic acid sequence endogenous to the subject. For example, the nucleic acid sequence of IGFBP2 derived from humans can be used to express IGFBP2 in humans. In other examples, the nucleic acid sequence of the antigenic epitope can be a nucleic acid sequence exogenous to the subject. For example, the nucleic acid sequence of IGFBP2 derived from non-humans can be used to express IGFBP2 in humans.

[0189] In some examples, the nucleic acid sequence for expressing the antigenic epitope can be a wild-type nucleic acid sequence. For example, the nucleic acid sequence of the natural origin of IGFBP2 in a certain genome can be used to express IGFBP2 in the subject. In other examples, the nucleic acid sequence encoding the epitope can be a synthetic nucleic acid sequence. For example, the nucleic acid sequence of the natural origin of IGFBP2 in a certain genome can be modified using molecular techniques known to those skilled in the art and can be used for the expression of IGFBP2 in the subject.

[0190] In some examples, the nucleic acid sequence encoding the epitope can be a nucleic acid sequence endogenous to the subject. For example, the nucleic acid sequence of IGFBP2 derived from humans can be used to express IGFBP2 in humans. In other examples, the nucleic acid sequence of the antigenic epitope can be a nucleic acid sequence exogenous to the subject. For example, the nucleic acid sequence of IGFBP2 derived from non-humans can be used to express IGFBP2 in humans.

[0191] In some examples, the nucleic acid sequence for expressing the antigenic epitope can be a wild-type nucleic acid sequence. For example, the nucleic acid sequence of the natural origin of IGFBP2 in a certain genome can be used to express IGFBP2 in the subject. In other examples, the nucleic acid sequence encoding the epitope can be a synthetic nucleic acid sequence. For example, the nucleic acid sequence of the natural origin of IGFBP2 in a certain genome can be modified using molecular techniques known to those skilled in the art and can be used for the expression of IGFBP2 in the subject.

[0192] In some examples, the nucleic acid sequence encoding the epitope can be a nucleic acid sequence endogenous to the subject. For example, the nucleic acid sequence of IGF-1R derived from humans can be used to express IGF-1R in humans. In other examples, the nucleic acid sequence of the antigenic epitope can be a nucleic acid sequence exogenous to the subject. For example, the nucleic acid sequence of IGF-1R derived from non-humans can be used to express IGF-1R in humans.

[0193] In some examples, the nucleic acid sequence for expressing the antigenic epitope can be a wild-type nucleic acid sequence. For example, the nucleic acid sequence of natural origin of IGF-1R in a certain genome can be used to express IGF-1R in a subject. In other examples, the nucleic acid sequence encoding the epitope can be a synthetic nucleic acid sequence. For example, the nucleic acid sequence of natural origin of IGF-1R in a certain genome can be modified using molecular techniques known to those skilled in the art and can be used for the expression of IGF-1R in a subject.

[0194] In some examples, the nucleic acid sequence encoding the epitope can be a nucleic acid sequence endogenous to the subject. For example, the nucleic acid sequence of IGF-1R derived from humans can be used to express IGF-1R in humans. In other examples, the nucleic acid sequence of the antigenic epitope can be a nucleic acid sequence exogenous to the subject. For example, the nucleic acid sequence of IGF-1R derived from non-humans can be used to express IGF-1R in humans.

[0195] In some examples, the nucleic acid sequence for expressing the antigenic epitope can be a wild-type nucleic acid sequence. For example, the nucleic acid sequence of natural origin of IGF-1R in a certain genome can be used to express IGF-1R in a subject. In other examples, the nucleic acid sequence encoding the epitope can be a synthetic nucleic acid sequence. For example, the nucleic acid sequence of natural origin of IGF-1R in a certain genome can be modified using molecular techniques known to those skilled in the art and can be used for the expression of IGF-1R in a subject.

[0196] In some examples, the nucleic acid sequence encoding the epitope can be a nucleic acid sequence endogenous to the subject. For example, the nucleic acid sequence of CDH3 derived from humans can be used to express CDH3 in humans. In other examples, the nucleic acid sequence of the antigenic epitope can be a nucleic acid sequence exogenous to the subject. For example, the nucleic acid sequence of CDH3 derived from non-humans can be used to express CDH3 in humans.

[0197] In some examples, the nucleic acid sequence for expressing the antigenic epitope can be a wild-type nucleic acid sequence. For example, the nucleic acid sequence of the natural origin of CDH3 in a certain genome can be used to express CDH3 in a subject. In other examples, the nucleic acid sequence encoding the epitope can be a synthetic nucleic acid sequence. For example, the nucleic acid sequence of the natural origin of CDH3 in a certain genome can be modified using molecular techniques known to those skilled in the art and used for the expression of CDH3 in a subject.

[0198] In some examples, the nucleic acid sequence encoding the epitope can be a nucleic acid sequence endogenous to the subject. For example, the nucleic acid sequence of CDH3 derived from humans can be used to express CDH3 in humans. In other examples, the nucleic acid sequence of the antigenic epitope can be a nucleic acid sequence exogenous to the subject. For example, the nucleic acid sequence of CDH3 derived from non-humans can be used to express CDH3 in humans.

[0199] In some examples, the nucleic acid sequence for expressing the antigenic epitope can be a wild-type nucleic acid sequence. For example, the nucleic acid sequence of the natural origin of CDH3 in a certain genome can be used to express CDH3 in a subject. In other examples, the nucleic acid sequence encoding the epitope can be a synthetic nucleic acid sequence. For example, the nucleic acid sequence of the natural origin of CDH3 in a certain genome can be modified using molecular techniques known to those skilled in the art and used for the expression of CDH3 in a subject.

[0200] The composition described in this specification comprises a first plasmid containing a first nucleotide sequence encoding a first epitope of a first antigen expressed by breast cancer-related cells, and a second nucleotide sequence encoding a second epitope of a second antigen expressed by breast cancer-related cells, wherein the first nucleotide sequence and the second nucleotide sequence are located in one or more plasmids. In some examples, the composition can comprise nucleic acids encoding epitopes derived from the following proteins: CD105, HIF1α, MDM2, Yb-1, SOX-2, HER-2, IGFBP2, IGF-1R, and CDH3.

[0201] In some examples, the composition is a composition comprising a first plasmid containing a first nucleotide sequence encoding a first epitope of a first antigen, wherein the first epitope is a portion of a peptide selected from CD105, Yb-1, SOX-2, CDH3, or MDM2, and the first nucleotide sequence can be located in a plasmid. In other examples, the composition is a composition comprising a first plasmid containing a first nucleotide sequence encoding a first epitope of a first antigen and a second nucleotide sequence encoding a second epitope of a second antigen, wherein the first and second epitopes are independently selected from CD105, Yb-1, SOX-2, CDH3, or MDM2, and the first nucleotide sequence and the second nucleotide sequence can be located in one or more plasmids.

[0202] In one example, the composition can include nucleic acids encoding epitopes from the following proteins, CD105, MDM2, Yb-1, SOX-2, and CDH3.In some examples, the composition has a nucleotide sequence having at least 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the nucleotide sequence of CAGAACGGCACCTGGCCCCGCGAGGTGCTGCTGGTGCTGTCCGTGAACTCCTCCGTGTTCCTGCACCTACAGGCCCTGGGCATCCCCCTGCACCTGGCCTACAACTCCTCCCTGGTGACCTTCCAGGAGCCCCCCGGCGTGAACACCACCGAGCTG (SEQ ID NO: 2); a nucleotide sequence having at least 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the nucleotide sequence of ACCGTGTTCATGCGCCTGAACATCATCTCCCCCGACCTGTCCGGCTGCACCTCCAAGGGCCTGGTGCTGCCCGCCGTGCTGGGCATCACCTTCGGCGCCTTCCTGATCGGCGCCCTGCTGACCGCCGCCCTGTGGTACATCTACTCCCACACCCGCTCCCCCTCCAAGCGCGAGCCCGTGGTGGCCGTGGCCGCCCCCGCCTCCTCCGAGTCCTCCTCCACCAACCACTCCATCGGCTCCACCCAGTCCACCCCCTGCTCCACCTCCTCCATGGCC (SEQ ID NO: 3); a nucleotide sequence having at least 50%, 60%, 70%。 , a nucleotide sequence having 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity; at least 50%, 60%, 70%, 8 with respect to the nucleotide sequence of ACCGTGTCCATGCGCCTGAACATCGTGTCCCCCGACCTGTCCGGCAAGGGCCTGGTGCTGCCCTCCGTGCTGGGCATCACCTTCGGCGCCTTCCTGATCGGCGCCCTGCTGACCGCCGCCCTGTGGTACATCTACTCCCACACCCGCGCCCCCTCCAAGCGCGAGCCCGTGGTGGCCGTGGCCGCCCCCGCCTCCTCCGAGTCCTCCTCCACCAACCACTCCATCGGCTCCACCCAGTCCACCCCCTGCTCCACCTCCTCCATGGCC (SEQ ID NO: 5) Nucleotide sequences having 0%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity; nucleotide sequences encoding amino acid sequences having at least 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the amino acid sequence of EARMLNASIVASFVELPL (SEQ ID NO: 6); nucleotide sequences encoding amino acid sequences having at least 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the amino acid sequence of QNGTWPREVLLVLSVNSSVFLHLQALGIPLHLAYNSSLVTFQEPPGVNTTEL (SEQ ID NO: 1); nucleotide sequences encoding amino acid sequences having at least 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the amino acid sequence of TVFMRLNIISPDLSGCTSKGLVLPAVLGITFGAFLIGALLTAALWYIYSHTRSPSKREPVVAVAAPASSESSSTNHSIGSTQSTPCSTSSMA (SEQ ID NO: 8); nucleotide sequences encoding amino acid sequences having at least 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the amino acid sequence of TVSMRLNIVSPDLSGKGLVLPSVLGITFGAFLIGALLTAALWYIYSHTRGPSKREPVVAVAAPASSESSSTNHSIGSTQSTPCSTSSMA (SEQ ID NO: 9);or a nucleic acid sequence encoding an epitope of peptide CD105 selected from the group consisting of nucleotide sequences encoding amino acid sequences having at least 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the amino acid sequence of TVSMRLNIVSPDLSGKGLVLPSVLGITFGAFLIGALLTAALWYIYSHTRAPSKREPVVAVAAPASSESSSTNHSIGSTQSTPCSTSSMA (SEQ ID NO: 10). In some examples, the composition comprises a nucleotide sequence having at least 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the nucleotide sequence of GGAGTGCCAGTGCAGGGCTCCAAGTACGCTGCCGACCGCAACCACTACCGCCGCTACCCACGCCGTCGCGGCCCACCCCGCAACTACCAGCAGAAC (SEQ ID NO: 11); a nucleotide sequence having at least 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the nucleotide sequence of GGCGTGCCCGTGCAGGGCTCCAAGTACGCCGCCGACCGCAACCACTACCGCCGCTACCCCCGCCGCCGCGGCCCCCCCCGCAACTACCAGCAGAAC (SEQ ID NO: 12); a nucleotide sequence having at least 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the nucleotide sequence of GGCGTGCCCGTGCAGGGCTCCAAGTACGCCGCCGACCGCAACCACTACCGCCGCTACCCCCGCCGCCGCGGCCCCCCCCGCAACTACCAGCAGAAC (SEQ ID NO: 12); a nucleotide sequence encoding an amino acid sequence having at least 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the amino acid sequence of EDVFVHQTAIKKNNPRK (SEQ ID NO: 14);A nucleotide sequence encoding an amino acid sequence having at least 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the amino acid sequence of YRRNFNYRRRRPEN (SEQ ID NO: 15); or a nucleotide sequence encoding an amino acid sequence having at least 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the amino acid sequence of GVPVQGSKYAADRNHYRRYPRRRGPPRNYQQN (SEQ ID NO: 16). In some examples, the composition can comprise at least with respect to the nucleotide sequence of GGCCTCAATGCGCACGGCGCAGCGCAGATGCAGCCCATGCACCGCTACGACGTGAGCGCCCTGCAGTACAACTCCATGACCAGCTCGCAGACCTACATGAACGGCTCGCCCACCTACAGCATGTCCTACTCGCAGCAGGGCACCCCTGGCATGGCTCTTGGCTCCATGGGTTCGGTG (SEQ ID NO: 17); A nucleotide sequence having at least 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity; at least 50%, 60 a nucleotide sequence having 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity; at least 50%, 60%, 70% with respect to the nucleotide sequence of GGCCTGAACGCCCACGGCGCCGCCCAGATGCAGCCCATGCACCGCTACGACGTGTCCGCCCTGCAGTACAACTCCATGACCTCCTCCCAGACCTACATGAACGGCTCCCCCACCTACTCCATGTCCTACTCCCAGCAGGGCACCCCCGGCATGGCCCTGGGCTCCATGGGCTCCGTG (SEQ ID NO: 18) A nucleotide sequence having 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity; or a nucleotide sequence encoding an amino acid sequence having at least 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the amino acid sequence of GLNAHGAAQMQPMHRYDVSALQYNSMTSSQTYMNGSPTYSMSYSQQGTPGMALGSMGSV (SEQ ID NO: 20). In some examples, the composition has at least 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the nucleotide sequence of AGGTCACTGAAGGAAAGGAATCCATTGAAAATCTTCCCATCCAAACGTATCTTACGAAGACACAAGAGAGATTGGGTGGTTGCTCCAATATCTGTCCCTGAAAATGGCAAGGGTCCCTTCCCACAGAGACTGAATCAGCTCAAGTCTAATAAAGATAGAGACACCAAGATTTTCTACAGCATCACGGGGCCGGGTGCAGACAGCCCACCTGAGGGTGTCTTCGCTGTAGAGAAGGAGACA (SEQ ID NO: 21); at least 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the nucleotide sequence of TTGAAAATCTTCCCATCCAAACGTATCTTACGAAGACACAAGAGAGATTGGGTGGTTGCTCCAATATCTGTCCCTGAAAATGGCAAGGGTCCCTTCCCACAGAGACTGAATCAGCTCAAGTCTAATAAAGATAGAGACACCAAGATTTTCTACAGCATCACGGGGCCGGGTGCAGACAGCCCACCTGAGGGTGTCTTCGCTGTAGAGAAGGAGACA (SEQ ID NO: 22);A nucleotide sequence having at least 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the nucleotide sequence of GCCATGCACTCCCCCCCCACCCGCATCCTGCGCCGCCGCAAGCGCGAGTGGGTGATGCCCCCCATCTTCGTGCCCGAGAACGGCAAGGGCCCCTTCCCCCAGCGCCTGAACCAGCTGAAGTCCAACAAGGACCGCGGCACCAAGATCTTCTACTCCATCACCGGCCCCGGCGCCGACTCCCCCCCCGAGGGCGTGTTCACCATCGAGAAGGAGTCC (SEQ ID NO: 23); A nucleotide sequence having at least 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the nucleotide sequence of GTGATGAACTCCCCCCCCTCCCGCATCCTGCGCCGCCGCAAGCGCGAGTGGGTGATGCCCCCCATCTCCGTGCCCGAGAACGGCAAGGGCCCCTTCCCCCAGCGCCTGAACCAGCTGAAGTCCAACAAGGACCGCGGCACCAAGCTGTTCTACTCCATCACCGGCCCCGGCGCCGACTCCCCCCCCGAGGGCGTGTTCACCATCGAGAAGGAGACC (SEQ ID NO: 24); A nucleotide sequence encoding an amino acid sequence having at least 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the amino acid sequence of RSLKERNPLKIFPSKRILRRHKRDWVVAPISVPENGKGPFPQRLNQLKSNKDRDTKIFYSITGPGADSPPEGVFAVEKET (SEQ ID NO: 25);A nucleotide sequence encoding an amino acid sequence having at least 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the amino acid sequence of LKIFPSKRILRRHKRDWVVAPISVPENGKGPFPQRLNQLKSNKDRDTKIFYSITGPGADSPPEGVFAVEKET (SEQ ID NO: 26); A nucleotide sequence encoding an amino acid sequence having at least 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the amino acid sequence of AMHSPPTRILRRRKREWVMPPIFVPENGKGPFPQRLNQLKSNKDRGTKIFYSITGPGADSPPEGVFTIEKES (SEQ ID NO: 27); or A nucleotide sequence encoding an amino acid sequence having at least 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the amino acid sequence of VMNSPPSRILRRRKREWVMPPISVPENGKGPFPQRLNQLKSNKDRGTKLFYSITGPGADSPPEGVFTIEKET (SEQ ID NO: 28), and can include a nucleic acid sequence encoding an epitope of peptide CDH3 selected from the group consisting of. In some examples, the composition is at least to the nucleotide sequence of ACCTACACCATGAAGGAGGTGCTGTTCTACCTGGGCCAGTACATCATGACCAAGCGCCTGTACGACGAGAAGCAGCAGCACATCGTGTACTGCTCCAACGACCTGCTGGGCGACCTGTTCGGCGTGCCCTCCTTCTCCGTGAAGGAGCACCGCAAaATCTACACCATGATCTACCGCAACCTGGTGGTGGTGAACCAGCAGGAGTCCTCCGACTCCGGCACCTCCGTGTCC (SEQ ID NO: 29) -encoding nucleotide sequence; or a nucleic acid sequence encoding an epitope of peptide CDH3 selected from the group consisting of a nucleotide sequence encoding an amino acid sequence having at least 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the amino acid sequence of VMNSPPSRILRRRKREWVMPPISVPENGKGPFPQRLNQLKSNKDRGTKLFYSITGPGADSPPEGVFTIEKET (SEQ ID NO: 28). In some examples, the composition is at least a nucleotide sequence having at least 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity; a nucleotide sequence having at least 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the nucleotide sequence of ACCTACACCATGAAGGAGATCATCTTCTACATCGGCCAGTACATCATGACCAAGCGCCTGTACGACGAGAAGCAGCAGCACATCGTGTACTGCTCCAACGACCTGCTGGGCGACGTGTTCGGCGTGCCCTCCTTCTCCGTGAAGGAGCACCGCAAGATCTACGCCATGATCTACCGCAACCTGGTGGCCGTGTCCCAGCAGGACTCCGGCACCTCCCTGTCC (SEQ ID NO: 30); a nucleotide sequence having at least 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the nucleotide sequence of ATCTACACCATGAAGGAGATCATCTTCTACATCGGCCAGTACATCATGACCAAGCGCCTGTACGACGAGAAGCAGCAGCACATCGTGTACTGCTCCAACGACCTGCTGGGCGACGTGTTCGGCGTGCCCTCCTTCTCCGTGAAGGAGCACCGCAAGATCTACGCCATGATCTACCGCAACCTGGTGGTGGTGTCCCAGCAGGACTCCGGCACCTCCCCCTCC (SEQ ID NO: 31); a nucleotide sequence encoding an amino acid sequence having at least 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the amino acid sequence of TYTMKEVLFYLGQYIMTKRLYDEKQQHIVYCSNDLLGDLFGVPSFSVKEHRKIYTMIYRNLVVVNQQESSDSGTSV (SEQ ID NO: 32);A nucleotide sequence encoding an amino acid sequence having at least 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the amino acid sequence of TYTMKEIIFYIGQYIMTKRLYDEKQQHIVYCSNDLLGDVFGVPSFSVKEHRKIYAMIYRNLVAVSQQDSGTSLS (SEQ ID NO: 33); or IYTMKEIIFYIGQYIMTKRLYDEKQQHIVYCSNDLLGDVFGVPSFSVKEHRKIYAMIYRNLVVVSQQDSGTSPS (SEQ ID NO: 34) A nucleic acid sequence encoding an epitope of the peptide MDM2 selected from the group consisting of nucleotide sequences encoding amino acid sequences having at least 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the amino acid sequence of can be included.;

[0203] In an exemplary example, the composition isATGGCGGTACCCATGCAACTGTCCTGCTCTAGACAGAACGGCACCTGGCCCCGCGAGGTGCTGCTGGTGCTGTCCGTGAACTCCTCCGTGTTCCTGCACCTACAGGCCCTGGGCATCCCCCTGCACCTGGCCTACAACTCCTCCCTGGTGACCTTCCAGGAGCCCCCCGGCGTGAACACCACCGAGCTGAGATCCACCGGTGGAGTGCCAGTGCAGGGCTCCAAGTACGCTGCCGACCGCAACCACTACCGCCGCTACCCACGCCGTCGCGGCCCACCCCGCAACTACCAGCAGAACACGCGTGGCCTCAATGCGCACGGCGCAGCGCAGATGCAGCCCATGCACCGCTACGACGTGAGCGCCCTGCAGTACAACTCCATGACCAGCTCGCAGACCTACATGAACGGCTCGCCCACCTACAGCATGTCCTACTCGCAGCAGGGCACCCCTGGCATGGCTCTTGGCTCCATGGGTTCGGTGAGATCCCAATTGAGGTCACTGAAGGAAAGGAATCCATTGAAAATCTTCCCATCCAAACGTATCTTACGAAGACACAAGAGAGATTGGGTGGTTGCTCCAATATCTGTCCCTGAAAATGGCAAGGGTCCCTTCCCACAGAGACTGAATCAGCTCAAGTCTAATAAAGATAGAGACACCAAGATTTTCTACAGCATCACGGGGCCGGGTGCAGACAGCCCACCTGAGGGTGTCTTCGCTGTAGAGAAGGAGACAAGATCCGCCGGCGAAACCTACACCATGAAGGAGGTGCTGTTCTACCTGGGCCAGTACATCATGACCAAGCGCCTGTACGACGAGAAGCAGCAGCACATCGTGTACTGCTCCAACGACCTGCTGGGCGACCTGTTCGGCGTGCCCTCCTTCTCCGTGAAGGAGCACCGCAAAATCTACACCATGATCTACCGCAACCTGGTGGTGGTGAACCAGCAGGAGTCCTCCGACTCCGGCACCTCCGTGTCCAGATCTTAG (SEQ ID NO: 35) nucleotides,A nucleotide sequence having at least 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the nucleotide sequence of (38); a nucleotide sequence encoding an amino acid sequence having at least 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the amino acid sequence of MAVPMQLSCSRQNGTWPREVLLVLSVNSSVFLHLQALGIPLHLAYNSSLVTFQEPPGVNTTELRSTGGVPVQGSKYAADRNHYRRYPRRRGPPRNYQQNTRGLNAHGAAQMQPMHRYDVSALQYNSMTSSQTYMNGSPTYSMSYSQQGTPGMALGSMGSVRSQLRSLKERNPLKIFPSKRILRRHKRDWVVAPISVPENGKGPFPQRLNQLKSNKDRDTKIFYSITGPGADSPPEGVFAVEKETRSAGETYTMKEVLFYLGQYIMTKRLYDEKQQHIVYCSNDLLGDLFGVPSFSVKEHRKIYTMIYRNLVVVNQQESSDSGTSVSRS (SEQ ID NO: 39);A nucleotide sequence encoding an amino acid sequence having at least 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the amino acid sequence of MAVPMTVFMRLNIISPDLSGCTSKGLVLPAVLGITFGAFLIGALLTAALWYIYSHTRSPSKREPVVAVAAPASSESSSTNHSIGSTQSTPCSTSSMATGGVPVQGSKYAADRNHYRRYPRRRGPPRNYQQNTRGLNAHGAAQMQPMHRYDVSALQYNSMTSSQTYMNGSPTYSMSYSQQGTPGMALGSMGSVRSQLLKIFPSKRILRRHKRDWVVAPISVPENGKGPFPQRLNQLKSNKDRDTKIFYSITGPGADSPPEGVFAVEKETRSAGETYTMKEVLFYLGQYIMTKRLYDEKQQHIVYCSNDLLGDLFGVPSFSVKEHRKIYTMIYRNLVVVNQQESSDSGTSVSRS (SEQ ID NO: 40); A nucleotide sequence encoding an amino acid sequence having at least 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the amino acid sequence of MAVPMTVSMRLNIVSPDLSGKGLVLPSVLGITFGAFLIGALLTAALWYIYSHTRGPSKREPVVAVAAPASSESSSTNHSIGSTQSTPCSTSSMATGGVPVQGSKYAADRNHYRRYPRRRGPPRNYQQNTRGLNAHGAAQMQPMHRYDVSALQYNSMTSSQTYMNGSPTYSMSYSQQGTPGMALGSMGSVRSQLAMHSPPTRILRRRKREWVMPPIFVPENGKGPFPQRLNQLKSNKDRGTKIFYSITGPGADSPPEGVFTIEKESRSAGETYTMKEIIFYIGQYIMTKRLYDEKQQHIVYCSNDLLGDVFGVPSFSVKEHRKIYAMIYRNLVAVSQQDSGTSLSRS (SEQ ID NO: 41);Or a nucleic acid sequence encoding a fusion peptide of five epitopes selected from the group consisting of nucleotide sequences encoding amino acid sequences having at least 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the amino acid sequence of MAVPMTVSMRLNIVSPDLSGKGLVLPSVLGITFGAFLIGALLTAALWYIYSHTRAPSKREPVVAVAAPASSESSSTNHSIGSTQSTPCSTSSMATGGVPVQGSKYAADRNHYRRYPRRRGPPRNYQQNTRGLNAHGAAQMQPMHRYDVSALQYNSMTSSQTYMNGSPTYSMSYSQQGTPGMALGSMGSVRSQLVMNSPPSRILRRRKREWVMPPISVPENGKGPFPQRLNQLKSNKDRGTKLFYSITGPGADSPPEGVFTIEKETRSAGEIYTMKEIIFYIGQYIMTKRLYDEKQQHIVYCSNDLLGDVFGVPSFSVKEHRKIYAMIYRNLVVVSQQDSGTSPSRS (SEQ ID NO: 42).;

[0204] In some examples, the composition can include nucleic acids encoding epitopes from the following proteins, HER-2, IGFBP2, and IGF-1R. In some examples, the composition can include a first plasmid comprising a first nucleotide sequence encoding a first epitope of a first antigen, the first epitope being a portion of a peptide selected from IGFBP-2, HER-2, IGF-1R, and the first nucleotide sequence being located in the plasmid. In some examples, the composition can include a first plasmid comprising a first nucleotide sequence encoding a first epitope of a first antigen and a second nucleotide sequence encoding a second epitope of a second antigen, the first and second epitopes being independently selected from IGFBP-2, HER-2, or IGF-1R, and the first nucleotide sequence and the second nucleotide sequence being located in one or more plasmids. In some examples, the composition is at least 50% relative to the nucleotide sequence of ATGCTGCCGAGAGTGGGCTGCCCCGCGCTGCCGCTGCCGCCGCCGCCGCTGCTGCCGCTGCTGCCGCTGCTGCTGCTGCTACTGGGCGCGAGTGGCGGCGGCGGCGGGGCGCGCGCGGAGGTGCTGTTCCGCTGCCCGCCCTGCACACCCGAGCGCCTGGCCGCCTGCGGGCCCCCGCCGGTTGCGCCGCCCGCCGCGGTGGCCGCAGTGGCCGGAGGCGCCCGCATGCCATGCGCGGAGCTCGTCCGGGAGCCGGGCTGCGGCTGCTGCTCGGTGTGCGCCCGGCTGGAGGGCGAGGCGTGCGGCGTCTACACCCCGCGCTGCGGCCAGGGGCTGCGCTGCTATCCCCACCCGGGCTCCGAGCTGCCCCTGCAGGCGCTGGTCATGGGCGAGGGCACTTGTGAGAAGCGCCGGGACGCCGAGTATGGCGCCAGCCCGGAGCAGGTTGCAGACAATGGCGATGACCACTCAGAAGGAGGCCTGGTGGAG (SEQ ID NO: 43), 6 Nucleotide sequences having 0%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity; at least 50%, 60% with respect to the nucleotide sequence of ATGCTGCCCCGCCTGGGCGGCCCCGCCCTGCCCCTGCTGCTGCCCTCCCTGCTGCTGCTGCTGCTGCTGGGCGCCGGCGGCTGCGGCCCCGGCGTGCGCGCCGAGGTGCTGTTCCGCTGCCCCCCCTGCACCCCCGAGCGCCTGGCCGCCTGCGGCCCCCCCCCCGACGCCCCCTGCGCCGAGCTGGTGCGCGAGCCCGGCTGCGGCTGCTGCTCCGTGTGCGCCCGCCAGGAGGGCGAGGCCTGCGGCGTGTACATCCCCCGCTGCGCCCAGACCCTGCGCTGCTACCCCAACCCCGGCTCCGAGCTGCCCCTGAAGGCCCTGGTGACCGGCGCCGGCACCTGCGAGAAGCGCCGCGTGGGCACCACCCCCCAGCAGGTGGCCGACTCCGACGACGACCACTCCGAGGGCGGCCTGGTGGAG (SEQ ID NO: 44) Nucleotide sequences having 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity; at least 50%, 60%, 70% with respect to the nucleotide sequence of ATGCTGCCCCGCCTGGGCGGCCCCGCCCTGCCCCTGCTGCTGCCCTCCCTGCTGCTGCTGCTGCTGCTGGGCGCCGGCGGCTGCGGCCCCGGCGTGCGCGCCGAGGTGCTGTTCCGCTGCCCCCCCTGCACCCCCGAGCGCCTGGCCGCCTGCGGCCCCCCCCCCGACGCCCCCTGCGCCGAGCTGGTGCGCGAGCCCGGCTGCGGCTGCTGCTCCGTGTGCGCCCGCCAGGAGGGCGAGGCCTGCGGCGTGTACATCCCCCGCTGCGCCCAGACCCTGCGCTGCTACCCCAACCCCGGCTCCGAGCTGCCCCTGAAGGCCCTGGTGACCGGCGCCGGCACCTGCGAGAAGCGCCGCGTGGGCACCACCCCCCAGCAGGTGGCCGACTCCGAGGACGACCACTCCGAGGGCGGCCTGGTGGAG (SEQ ID NO: 45) A nucleotide sequence having at least 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to an array; a nucleotide sequence encoding an amino acid sequence having at least 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the amino acid sequence of TMRRLLQETELVEPLTPSGAMPNQAQMRILKETELRKVKVLGSGAFGTVYKGIWIPDGENVKIPVAIKVLRENTSPKANKEILDEAYVMAGVGSPYVSRLLGICLTSTVQLVTQLMPYGCLLDHVRENRGRLGSQDLLNWCMQIAKGMSYLEDVRLVHRDLAARNVLVKSPNHVKITDFGLARLLDIDETEYHADGGKVPIKWMALESILRRRFTHQSDVWSYGVTVWELMTFGAKPYDGIPAREIPDLLEKGERLPQPPICTIDVYMIMVKCWMIDSECRPRFRELVSEFSRMARDPQRFVVIQNEDLAPGAGGMVHHRHRSSSPLPAARPAGATLERPKTLSPGKNGVVKDVFAFGGAVENPEYL (SEQ ID NO: 60); at least 50% with respect to the amino acid sequence of TMRRLLQETELVEPLTPSGAVPNQAQMRILKETELRKLKVLGSGAFGTVYKGIWIPDGENVKIPVAIKVLRENTSPKANKEILDEAYVMAGVGSPYVSRLLGICLTSTVQLVTQLMPYGCLLDHVREHRGRLGSQDLLNWCVQIAKGMSYLEEVRLVHRDLAARNVLVKSPNHVKITDFGLARLLDIDETEYHADGGKVPIKWMALESILRRRFTHQSDVWSYGVTVWELMTFGAKPYDGIPAREIPDLLEKGERLPQPPICTIDVYMIMVKCWMIDSECRPRFRELVSEFSRMARDPQRFVVIQNEDLALGTGSTAHRRHRSSSPPPPIRPAGATLERPKTLSPGKNGVVKDVFAFGGAVENPEYL (SEQ ID NO: 61), A nucleotide sequence encoding an amino acid sequence having 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity; and TMRRLLQETELVEPLTPSGAMPNQAQMRILKETELRKVKVLGSGAFGTVYKGIWIPDGENVKIPVAIKVLRENTSPKANKEILDEAYVMAGVGSPYVSRLLGICLTSTVQLVTQLMPYGCLLDHVREHRGRLGSQDLLNWCVQIAKGMSYLEDVRLVHRDLAARNVLVKSPNHVKITDFGLARLLDIDETEYHADGGKVPIKWMALESILRRRFTHQSDVWSYGVTVWELMTFGAKPYDGIPAREIPDLLEKGERLPQPPICTIDVYMIMVKCWMIDSECRPRFRELVSEFSRMARDPQRFVVIQNEDLTPGTGSTAHRRHRSSSPLPPVRPAGATLERPKTLSPGKNGVVKDVFAFGGAVENPEYL (SEQ ID NO At least 50%, 60%, 70%, 80%, 90%, 91%, 92%, 9 A nucleotide sequence having at least 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the nucleotide sequence of (78); A nucleotide sequence encoding an amino acid sequence having at least 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the amino acid sequence of MAVPMLPRVGCPALPLPPPPLLPLLPLLLLLLGASGGGGGARAEVLFRCPPCTPERLAACGPPPVAPPAAVAAVAGGARMPCAELVREPGCGCCSVCARLEGEACGVYTPRCGQGLRCYPHPGSELPLQALVMGEGTCEKRRDAEYGASPEQVADNGDDHSEGGLVEQLTMRRLLQETELVEPLTPSGAMPNQAQMRILKETELRKVKVLGSGAFGTVYKGIWIPDGENVKIPVAIKVLRENTSPKANKEILDEAYVMAGVGSPYVSRLLGICLTSTVQLVTQLMPYGCLLDHVRENRGRLGSQDLLNWCMQIAKGMSYLEDVRLVHRDLAARNVLVKSPNHVKITDFGLARLLDIDETEYHADGGKVPIKWMALESILRRRFTHQSDVWSYGVTVWELMTFGAKPYDGIPAREIPDLLEKGERLPQPPICTIDVYMIMVKCWMIDSECRPRFRELVSEFSRMARDPQRFVVIQNEDLAPGAGGMVHHRHRSSSPLPAARPAGATLERPKTLSPGKNGVVKDVFAFGGAVENPEYLGRPVPWSFGVVLWEIATLAEQPYQGLSNEQVLRFVMEGGLLDKPDNCPDMLFELMRMCWQYNPKMRPSFLEHKAENGPGPGVLVLRASFDERQPYAHMNGGRKNERALPAAA (SEQ ID NO: 79);A nucleotide sequence encoding an amino acid sequence having at least 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the amino acid sequence of MAVPMLPRLGGPALPLLLPSLLLLLLLGAGGCGPGVRAEVLFRCPPCTPERLAACGPPPDAPCAELVREPGCGCCSVCARQEGEACGVYIPRCAQTLRCYPNPGSELPLKALVTGAGTCEKRRVGTTPQQVADSDDDHSEGGLVEQLTMRRLLQETELVEPLTPSGAVPNQAQMRILKETELRKLKVLGSGAFGTVYKGIWIPDGENVKIPVAIKVLRENTSPKANKEILDEAYVMAGVGSPYVSRLLGICLTSTVQLVTQLMPYGCLLDHVREHRGRLGSQDLLNWCVQIAKGMSYLEEVRLVHRDLAARNVLVKSPNHVKITDFGLARLLDIDETEYHADGGKVPIKWMALESILRRRFTHQSDVWSYGVTVWELMTFGAKPYDGIPAREIPDLLEKGERLPQPPICTIDVYMIMVKCWMIDSECRPRFRELVSEFSRMARDPQRFVVIQNEDLALGTGSTAHRRHRSSSPPPPIRPAGATLERPKTLSPGKNGVVKDVFAFGGAVENPEYLGRPVPWSFGVVLWEIATLAEQPYQGLSNEQVLRFVMEGGLLDKPDNCPDMLFELMRMCWQYNPKMRPSFLEHKAENGPGPGVLVLRASFDERQPYAHMNGGRANERALPAAA (SEQ ID NO: 80);and a nucleic acid sequence encoding a fusion protein of three epitopes selected from the group consisting of nucleotide sequences encoding amino acid sequences having at least 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the amino acid sequence of MAVPMLPRLGGPALPLLLPSLLLLLLLGAGGCGPGVRAEVLFRCPPCTPERLAACGPPPDAPCAELVREPGCGCCSVCARQEGEACGVYIPRCAQTLRCYPNPGSELPLKALVTGAGTCEKRRVGATPQQVADSEDDHSEGGLVEQLTMRRLLQETELVEPLTPSGAMPNQAQMRILKETELRKVKVLGSGAFGTVYKGIWIPDGENVKIPVAIKVLRENTSPKANKEILDEAYVMAGVGSPYVSRLLGICLTSTVQLVTQLMPYGCLLDHVREHRGRLGSQDLLNWCVQIAKGMSYLEDVRLVHRDLAARNVLVKSPNHVKITDFGLARLLDIDETEYHADGGKVPIKWMALESILRRRFTHQSDVWSYGVTVWELMTFGAKPYDGIPAREIPDLLEKGERLPQPPICTIDVYMIMVKCWMIDSECRPRFRELVSEFSRMARDPQRFVVIQNEDLTPGTGSTAHRRHRSSSPLPPVRPAGATLERPKTLSPGKNGVVKDVFAFGGAVENPEYLGRPVPWSFGVVLWEIATLAEQPYQGLSNEQVLRFVMEGGLLDKPDNCPDMLFELMRMCWQYNPKMRPSFLEHKAENGPGVLVLRASFDERQPYAHMNGGRANERALPAAA (SEQ ID NO: 81).;

[0205] In some examples, the composition can include first and second epitopes independently selected from CD105, Yb-1, SOX-2, CDH3, or MDM2. In some examples, the composition can include a third epitope, and the first, second, and third epitopes are independently selected from CD105, Yb-1, SOX-2, CDH3, or MDM2. In some examples, the composition can include third and fourth epitopes, and the first, second, third, and fourth epitopes are independently selected from CD105, Yb-1, SOX-2, CDH3, or MDM2. In some examples, the composition can include third, fourth, and fifth epitopes, and the first, second, third, fourth, and fifth epitopes are independently selected from CD105, Yb-1, SOX-2, CDH3, or MDM2.

[0206] In some examples, the composition can include first and second epitopes independently selected from IGFBP2, HER-2, or IGF-1R. In some examples, the composition can include a third epitope, and the first, second, and third epitopes are independently selected from IGFBP2, HER-2, or IGF-1R.

[0207] In some examples, the composition can be administered to a subject. In some examples, the subject is in need of administration of the composition. In some examples, the composition is effective in inducing an immune response in the subject. In some examples, the composition is effective in eliminating a number of cells associated with breast cancer in the subject. In some examples, the composition can be used to prevent the growth of cells associated with breast cancer in the subject.

[0208] In some examples, the first and second nucleic acid sequences are located on a first plasmid. In some examples, the second nucleic acid sequence is located on a second plasmid.

[0209] In some examples, the cells associated with breast cancer are selected from breast cells expressing atypical features, pre-tumor breast cells, breast cancer cells, pre-invasive breast cancer cells, breast cancer stem cells, epithelial cells, mesenchymal cells, stromal cells, or combinations thereof.

[0210] In some examples, the first and second nucleic acid sequences are purified to at least 70% purity. In some examples, the first and second nucleic acid sequences are located on a first plasmid and are separated by the sequence of a linker nucleic acid. In some examples, the first nucleic acid sequence is adjacent to the second nucleic acid sequence in the first plasmid.

[0211] In some examples, at least the first plasmid is contained within a pharmaceutical composition. In some examples, at least the first plasmid is contained within a pharmaceutical composition that further comprises a pharmaceutical carrier. In some examples, at least the first plasmid is contained within a pharmaceutical composition that further comprises a pharmaceutical carrier and an adjuvant. In some examples, at least the first plasmid is contained within a pharmaceutical composition that further comprises an adjuvant. In some examples, the composition further comprises an adjuvant and a pharmaceutically acceptable carrier. In some examples, the adjuvant is GM-CSF.

[0212] In some examples, the subject is selected from a human having breast cancer, a mouse having breast cancer, or a rat having breast cancer. In some examples, the subject is selected from a human without breast cancer, a mouse without breast cancer, or a rat without breast cancer.

[0213] In some examples, the immune response is a type 1 immune response. In some examples, the first nucleic acid sequence is a species selected from human, mouse, or rat. In some examples, the second nucleic acid sequence is a species selected from human, mouse, or rat. In some examples, the immune response is characterized by the ratio of type I cytokine production to type II cytokine production that is greater than 1. In some examples, the immune response is characterized by the ratio of type I cytokine production to type II cytokine production that is less than 1. In some examples, the immune response is characterized by the ratio of IFNγ production to IL-10 production that is greater than 1. In some examples, the immune response is characterized by the ratio of IFNγ production to IL-10 production that is less than 1.

[0214] In some examples, the composition is a composition comprising a first plasmid comprising a first nucleotide sequence encoding a first epitope of a first antigen, wherein the first epitope is a portion of the HIF-1α peptide, and the first nucleotide sequence is located in the plasmid. In other examples, the composition is a composition comprising a first plasmid comprising a first nucleotide sequence encoding a first epitope of a first antigen and a second nucleotide sequence encoding a second epitope of a second antigen, wherein the first and second epitopes are portions of the HIF-1α peptide, and the first nucleotide sequence and the second nucleotide sequence are located in one or more plasmids.

[0215] Nucleic acid sequences encoding epitopes from the following proteins, CD105, HIF1α, MDM2, Yb-1, SOX-2, HER-2, IGFBP2, IGF-1R and CDH3, can be different from those described herein. In some examples, nucleic acid sequences that are more than 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% identical to the nucleic acid sequences disclosed herein, or more than 50% identical, can be used in the compositions described herein.

[0216] In some examples, the compositions described herein can include compositions that include a first epitope of a first antigen expressed by cells associated with breast cancer and a second epitope of a second antigen expressed by cells associated with breast cancer.

[0217] In some examples, the composition can comprise at least a first epitope of a first antigen, the first epitope being a portion of a peptide selected from CD105, Yb-1, SOX-2, CDH3 or MDM2. In some examples, the composition can comprise at least a first epitope of a first antigen and at least a second epitope of a second antigen, the first and second epitopes being independently selected from CD105, Yb-1, SOX-2, CDH3 or MDM2.In some examples, at least the first epitope of the peptide CD105 is an amino acid sequence having at least 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the amino acid sequence of EARMLNASIVASFVELPL (SEQ ID NO: 6); an amino acid sequence having at least 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the amino acid sequence of QNGTWPREVLLVLSVNSSVFLHLQALGIPLHLAYNSSLVTFQEPPGVNTTEL (SEQ ID NO: 1); an amino acid sequence having at least 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the amino acid sequence of TVFMRLNIISPDLSGCTSKGLVLPAVLGITFGAFLIGALLTAALWYIYSHTRSPSKREPVVAVAAPASSESSSTNHSIGSTQSTPCSTSSMA (SEQ ID NO: 8); an amino acid sequence having at least 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the amino acid sequence of TVSMRLNIVSPDLSGKGLVLPSVLGITFGAFLIGALLTAALWYIYSHTRGPSKREPVVAVAAPASSESSSTNHSIGSTQSTPCSTSSMA (SEQ ID NO: 9); or an amino acid sequence having at least 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the amino acid sequence of TVSMRLNIVSPDLSGKGLVLPSVLGITFGAFLIGALLTAALWYIYSHTRAPSKREPVVAVAAPASSESSSTNHSIGSTQSTPCSTSSMA (SEQ ID NO: 10), and is selected from the group consisting of these amino acid sequences.In some examples, at least the first epitope of peptide Yb-1 is an amino acid sequence having at least 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the amino acid sequence of EDVFVHQTAIKKNNPRK (SEQ ID NO: 14); an amino acid sequence having at least 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the amino acid sequence of YRRNFNYRRRRPEN (SEQ ID NO: 15); or an amino acid sequence having at least 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the amino acid sequence of GVPVQGSKYAADRNHYRRYPRRRGPPRNYQQN (SEQ ID NO: 16), and is selected from the group consisting of. In some examples, at least the first epitope of peptide SOX-2 is selected from the group consisting of amino acid sequences having at least 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the amino acid sequence of GLNAHGAAQMQPMHRYDVSALQYNSMTSSQTYMNGSPTYSMSYSQQGTPGMALGSMGSV (SEQ ID NO: 20).In some examples, at least the first epitope of the peptide CDH3 is an amino acid sequence having at least 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the amino acid sequence of RSLKERNPLKIFPSKRILRRHKRDWVVAPISVPENGKGPFPQRLNQLKSNKDRDTKIFYSITGPGADSPPEGVFAVEKET (SEQ ID NO: 25); an amino acid sequence having at least 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the amino acid sequence of LKIFPSKRILRRHKRDWVVAPISVPENGKGPFPQRLNQLKSNKDRDTKIFYSITGPGADSPPEGVFAVEKET (SEQ ID NO: 26); an amino acid sequence having at least 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the amino acid sequence of AMHSPPTRILRRRKREWVMPPIFVPENGKGPFPQRLNQLKSNKDRGTKIFYSITGPGADSPPEGVFTIEKES (SEQ ID NO: 27); or an amino acid sequence having at least 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the amino acid sequence of VMNSPPSRILRRRKREWVMPPISVPENGKGPFPQRLNQLKSNKDRGTKLFYSITGPGADSPPEGVFTIEKET (SEQ ID NO: 28), and is selected from the group consisting of.In some examples, at least the first epitope of the peptide MDM-2 is an amino acid sequence having at least 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the amino acid sequence of TYTMKEVLFYLGQYIMTKRLYDEKQQHIVYCSNDLLGDLFGVPSFSVKEHRKIYTMIYRNLVVVNQQESSDSGTSV (SEQ ID NO: 32); an amino acid sequence having at least 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the amino acid sequence of TYTMKEIIFYIGQYIMTKRLYDEKQQHIVYCSNDLLGDVFGVPSFSVKEHRKIYAMIYRNLVAVSQQDSGTSLS (SEQ ID NO: 33); or an amino acid sequence having at least 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the amino acid sequence of IYTMKEIIFYIGQYIMTKRLYDEKQQHIVYCSNDLLGDVFGVPSFSVKEHRKIYAMIYRNLVVVSQQDSGTSPS (SEQ ID NO: 34), and is selected from the group consisting of such amino acid sequences.

[0218] In one example, the compositions described herein have at least 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the amino acid sequence of MAVPMQLSCSRQNGTWPREVLLVLSVNSSVFLHLQALGIPLHLAYNSSLVTFQEPPGVNTTELRSTGGVPVQGSKYAADRNHYRRYPRRRGPPRNYQQNTRGLNAHGAAQMQPMHRYDVSALQYNSMTSSQTYMNGSPTYSMSYSQQGTPGMALGSMGSVRSQLRSLKERNPLKIFPSKRILRRHKRDWVVAPISVPENGKGPFPQRLNQLKSNKDRDTKIFYSITGPGADSPPEGVFAVEKETRSAGETYTMKEVLFYLGQYIMTKRLYDEKQQHIVYCSNDLLGDLFGVPSFSVKEHRKIYTMIYRNLVVVNQQESSDSGTSVSRS (SEQ ID NO: 39); at least 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the amino acid sequence of MAVPMTVFMRLNIISPDLSGCTSKGLVLPAVLGITFGAFLIGALLTAALWYIYSHTRSPSKREPVVAVAAPASSESSSTNHSIGSTQSTPCSTSSMATGGVPVQGSKYAADRNHYRRYPRRRGPPRNYQQNTRGLNAHGAAQMQPMHRYDVSALQYNSMTSSQTYMNGSPTYSMSYSQQGTPGMALGSMGSVRSQLLKIFPSKRILRRHKRDWVVAPISVPENGKGPFPQRLNQLKSNKDRDTKIFYSITGPGADSPPEGVFAVEKETRSAGETYTMKEVLFYLGQYIMTKRLYDEKQQHIVYCSNDLLGDLFGVPSFSVKEHRKIYTMIYRNLVVVNQQESSDSGTSVSRS (SEQ ID NO: 40);An amino acid sequence having at least 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the amino acid sequence of MAVPMTVSMRLNIVSPDLSGKGLVLPSVLGITFGAFLIGALLTAALWYIYSHTRGPSKREPVVAVAAPASSESSSTNHSIGSTQSTPCSTSSMATGGVPVQGSKYAADRNHYRRYPRRRGPPRNYQQNTRGLNAHGAAQMQPMHRYDVSALQYNSMTSSQTYMNGSPTYSMSYSQQGTPGMALGSMGSVRSQLAMHSPPTRILRRRKREWVMPPIFVPENGKGPFPQRLNQLKSNKDRGTKIFYSITGPGADSPPEGVFTIEKESRSAGETYTMKEIIFYIGQYIMTKRLYDEKQQHIVYCSNDLLGDVFGVPSFSVKEHRKIYAMIYRNLVAVSQQDSGTSLSRS (SEQ ID NO: 41); or an amino acid sequence having at least 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the amino acid sequence of MAVPMTVSMRLNIVSPDLSGKGLVLPSVLGITFGAFLIGALLTAALWYIYSHTRAPSKREPVVAVAAPASSESSSTNHSIGSTQSTPCSTSSMATGGVPVQGSKYAADRNHYRRYPRRRGPPRNYQQNTRGLNAHGAAQMQPMHRYDVSALQYNSMTSSQTYMNGSPTYSMSYSQQGTPGMALGSMGSVRSQLVMNSPPSRILRRRKREWVMPPISVPENGKGPFPQRLNQLKSNKDRGTKLFYSITGPGADSPPEGVFTIEKETRSAGEIYTMKEIIFYIGQYIMTKRLYDEKQQHIVYCSNDLLGDVFGVPSFSVKEHRKIYAMIYRNLVVVSQQDSGTSPSRS (SEQ ID NO: 42). The amino acid sequence of the fusion peptide of 5 epitopes selected from the group consisting of;

[0219] The composition described in this specification is a composition comprising a first plasmid containing a first nucleotide sequence encoding a first epitope of a first antigen, wherein the first epitope is a portion of a peptide selected from IGFBP-2, HER-2 or IGF-1R, and the first nucleotide sequence can comprise a composition located on the plasmid. In some examples, the composition can comprise a first plasmid containing a first nucleotide sequence encoding a first epitope of a first antigen and a second nucleotide sequence encoding a second epitope of a second antigen, wherein the first and second epitopes are independently selected from IGFBP-2, HER-2 or IGF-1R, and the first nucleotide sequence and the second nucleotide sequence are located on one or more plasmids. In some examples, at least the first epitope of the peptide IGFBP-2 has an amino acid sequence having at least 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the amino acid sequence of NHVDSTMNMLGGGGS (SEQ ID NO: 46); an amino acid sequence having at least 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the amino acid sequence of ELAVFREKVTEQHRQ (SEQ ID NO: 47); an amino acid sequence having at least 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the amino acid sequence of LGLEEPKKLRPPPAR (SEQ ID NO: 48); an amino acid sequence having at least 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the amino acid sequence of DQVLERISTMRLPDE (SEQ ID NO: 49); an amino acid sequence having at least 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the amino acid sequence of GPLEHLYSLHIPNCD (SEQ ID NO: 50);An amino acid sequence having at least 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the amino acid sequence of KHGLYNLKQCKMSLN (SEQ ID NO: 51); an amino acid sequence having at least 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the amino acid sequence of PECHLFYNEQQEARG (SEQ ID NO: 53); an amino acid sequence having at least 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the amino acid sequence of MLPRVGCPALPLPPPPLLPLLPLLLLLLGASGGGGGARAEVLFRCPPCTPERLAACGPPPVAPPAAVAAVAGGARMPCAELVREPGCGCCSVCARLEGEACGVYTPRCGQGLRCYPHPGSELPLQALVMGEGTCEKRRDAEYGASPEQVADNGDDHSEGGLVE (SEQ ID NO: 54); an amino acid sequence having at least 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the amino acid sequence of MLPRLGGPALPLLLPSLLLLLLLGAGGCGPGVRAEVLFRCPPCTPERLAACGPPPDAPCAELVREPGCGCCSVCARQEGEACGVYIPRCAQTLRCYPNPGSELPLKALVTGAGTCEKRRVGTTPQQVADSDDDHSEGGLVE (SEQ ID NO: 55);Or it is selected from the group consisting of amino acid sequences having at least 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the amino acid sequence of or MLPRLGGPALPLLLPSLLLLLLLGAGGCGPGVRAEVLFRCPPCTPERLAACGPPPDAPCAELVREPGCGCCSVCARQEGEACGVYIPRCAQTLRCYPNPGSELPLKALVTGAGTCEKRRVGATPQQVADSEDDHSEGGLVE (SEQ ID NO: 56). In some examples, at least the first epitope of the peptide HER-2 is or TMRRLLQETELVEPLTPSGAMPNQAQMRILKETELRKVKVLGSGAFGTVYKGIWIPDGENVKIPVAIKVLRENTSPKANKEILDEAYVMAGVGSPYVSRLLGICLTSTVQLVTQLMPYGCLLDHVRENRGRLGSQDLLNWCMQIAKGMSYLEDVRLVHRDLAARNVLVKSPNHVKITDFGLARLLDIDETEYHADGGKVPIKWMALESILRRRFTHQSDVWSYGVTVWELMTFGAKPYDGIPAREIPDLLEKGERLPQPPICTIDVYMIMVKCWMIDSECRPRFRELVSEFSRMARDPQRFVVIQNEDLAPGAGGMVHHRHRSSSPLPAARPAGATLERPKTLSPGKNGVVKDVFAFGGAVENPEYL (SEQ ID NO: 60) with at least 50%, 6; A nucleotide sequence encoding an amino acid sequence having 0%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity; TMRRLLQETELVEPLTPSGAVPNQAQMRILKETELRKLKVLGSGAFGTVYKGIWIPDGENVKIPVAIKVLRENTSPKANKEILDEAYVMAGVGSPYVSRLLGICLTSTVQLVTQLMPYGCLLDHVREHRGRLGSQDLLNWCVQIAKGMSYLEEVRLVHRDLAARNVLVKSPNHVKITDFGLARLLDIDETEYHADGGKVPIKWMALESILRRRFTHQSDVWSYGVTVWELMTFGAKPYDGIPAREIPDLLEKGERLPQPPICTIDVYMIMVKCWMIDSECRPRFRELVSEFSRMARDPQRFVVIQNEDLALGTGSTAHRRHRSSSPPPPIRPAGATLERPKTLSPGKNGVVKDVFAFGGAVENPEYL (SEQ ID NO: 61) A nucleotide sequence encoding an amino acid sequence having at least 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the amino acid sequence; or at least 50% to the amino acid sequence of TMRRLLQETELVEPLTPSGAMPNQAQMRILKETELRKVKVLGSGAFGTVYKGIWIPDGENVKIPVAIKVLRENTSPKANKEILDEAYVMAGVGSPYVSRLLGICLTSTVQLVTQLMPYGCLLDHVREHRGRLGSQDLLNWCVQIAKGMSYLEDVRLVHRDLAARNVLVKSPNHVKITDFGLARLLDIDETEYHADGGKVPIKWMALESILRRRFTHQSDVWSYGVTVWELMTFGAKPYDGIPAREIPDLLEKGERLPQPPICTIDVYMIMVKCWMIDSECRPRFRELVSEFSRMARDPQRFVVIQNEDLTPGTGSTAHRRHRSSSPLPPVRPAGATLERPKTLSPGKNGVVKDVFAFGGAVENPEYL (SEQ ID NO: 62) It is selected from the group consisting of nucleotide sequences encoding amino acid sequences having 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity. In some examples, the nucleic acid sequence encoding the epitope of the peptide IGF-1R has at least 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the amino acid sequence of DYRSYRFPKLTVITE (SEQ ID NO: 66); at least 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the amino acid sequence of IRGWKLFYNYALVIF (SEQ ID NO: 67); at least 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the amino acid sequence of VVTGYVKIRHSHALV (SEQ ID NO: 68); at least 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the amino acid sequence of FFYVQAKTGYENFIH (SEQ ID NO: 69); at least 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the amino acid sequence of LIIALPVAVLLIVGG (SEQ ID NO: 70); at least 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the amino acid sequence of LVIMLYVFHRKRNNS (SEQ ID NO: 71); at least 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the amino acid sequence of NCHHVVRLLGVVSQG (SEQ ID NO: 72);An amino acid sequence having at least 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the amino acid sequence of WSFGVVLWEIATLAEQPYQGLSNEQVLRFVMEGGLLDKPDNCPDMLFELMRMCWQYNPKMRPSFLEHKAENGPGPGVLVLRASFDERQPYAHMNGGRKNERALP (SEQ ID NO: 73); an amino acid sequence having at least 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the amino acid sequence of WSFGVVLWEIATLAEQPYQGLSNEQVLRFVMEGGLLDKPDNCPDMLFELMRMCWQYNPKMRPSFLEHKAENGPGPGVLVLRASFDERQPYAHMNGGRANERALP (SEQ ID NO: 74); or an amino acid sequence having at least 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the amino acid sequence of WSFGVVLWEIATLAEQPYQGLSNEQVLRFVMEGGLLDKPDNCPDMLFELMRMCWQYNPKMRPSFLEHKAENGPGVLVLRASFDERQPYAHMNGGRANERALP (SEQ ID NO: 75), selected from the group consisting of;

[0220] The compositions described herein have an amino acid sequence with at least 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the amino acid sequence of MAVPMLPRVGCPALPLPPPPLLPLLPLLLLLLGASGGGGGARAEVLFRCPPCTPERLAACGPPPVAPPAAVAAVAGGARMPCAELVREPGCGCCSVCARLEGEACGVYTPRCGQGLRCYPHPGSELPLQALVMGEGTCEKRRDAEYGASPEQVADNGDDHSEGGLVEQLTMRRLLQETELVEPLTPSGAMPNQAQMRILKETELRKVKVLGSGAFGTVYKGIWIPDGENVKIPVAIKVLRENTSPKANKEILDEAYVMAGVGSPYVSRLLGICLTSTVQLVTQLMPYGCLLDHVRENRGRLGSQDLLNWCMQIAKGMSYLEDVRLVHRDLAARNVLVKSPNHVKITDFGLARLLDIDETEYHADGGKVPIKWMALESILRRRFTHQSDVWSYGVTVWELMTFGAKPYDGIPAREIPDLLEKGERLPQPPICTIDVYMIMVKCWMIDSECRPRFRELVSEFSRMARDPQRFVVIQNEDLAPGAGGMVHHRHRSSSPLPAARPAGATLERPKTLSPGKNGVVKDVFAFGGAVENPEYLGRPVPWSFGVVLWEIATLAEQPYQGLSNEQVLRFVMEGGLLDKPDNCPDMLFELMRMCWQYNPKMRPSFLEHKAENGPGPGVLVLRASFDERQPYAHMNGGRKNERALPAAA (SEQ ID NO: 79);An amino acid sequence having at least 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the amino acid sequence of MAVPMLPRLGGPALPLLLPSLLLLLLLGAGGCGPGVRAEVLFRCPPCTPERLAACGPPPDAPCAELVREPGCGCCSVCARQEGEACGVYIPRCAQTLRCYPNPGSELPLKALVTGAGTCEKRRVGTTPQQVADSDDDHSEGGLVEQLTMRRLLQETELVEPLTPSGAVPNQAQMRILKETELRKLKVLGSGAFGTVYKGIWIPDGENVKIPVAIKVLRENTSPKANKEILDEAYVMAGVGSPYVSRLLGICLTSTVQLVTQLMPYGCLLDHVREHRGRLGSQDLLNWCVQIAKGMSYLEEVRLVHRDLAARNVLVKSPNHVKITDFGLARLLDIDETEYHADGGKVPIKWMALESILRRRFTHQSDVWSYGVTVWELMTFGAKPYDGIPAREIPDLLEKGERLPQPPICTIDVYMIMVKCWMIDSECRPRFRELVSEFSRMARDPQRFVVIQNEDLALGTGSTAHRRHRSSSPPPPIRPAGATLERPKTLSPGKNGVVKDVFAFGGAVENPEYLGRPVPWSFGVVLWEIATLAEQPYQGLSNEQVLRFVMEGGLLDKPDNCPDMLFELMRMCWQYNPKMRPSFLEHKAENGPGPGVLVLRASFDERQPYAHMNGGRANERALPAAA (SEQ ID NO: 80);Alternatively, it can further include a nucleic acid sequence encoding a fusion protein of three epitopes selected from the group consisting of amino acid sequences having at least 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the amino acid sequence of MAVPMLPRLGGPALPLLLPSLLLLLLLGAGGCGPGVRAEVLFRCPPCTPERLAACGPPPDAPCAELVREPGCGCCSVCARQEGEACGVYIPRCAQTLRCYPNPGSELPLKALVTGAGTCEKRRVGATPQQVADSEDDHSEGGLVEQLTMRRLLQETELVEPLTPSGAMPNQAQMRILKETELRKVKVLGSGAFGTVYKGIWIPDGENVKIPVAIKVLRENTSPKANKEILDEAYVMAGVGSPYVSRLLGICLTSTVQLVTQLMPYGCLLDHVREHRGRLGSQDLLNWCVQIAKGMSYLEDVRLVHRDLAARNVLVKSPNHVKITDFGLARLLDIDETEYHADGGKVPIKWMALESILRRRFTHQSDVWSYGVTVWELMTFGAKPYDGIPAREIPDLLEKGERLPQPPICTIDVYMIMVKCWMIDSECRPRFRELVSEFSRMARDPQRFVVIQNEDLTPGTGSTAHRRHRSSSPLPPVRPAGATLERPKTLSPGKNGVVKDVFAFGGAVENPEYLGRPVPWSFGVVLWEIATLAEQPYQGLSNEQVLRFVMEGGLLDKPDNCPDMLFELMRMCWQYNPKMRPSFLEHKAENGPGVLVLRASFDERQPYAHMNGGRANERALPAAA (SEQ ID NO: 81).;

[0221] In some examples, the composition comprises first and second epitopes independently selected from CD105, Yb-1, SOX-2, CDH3, or MDM2. In some examples, the composition further comprises a third epitope, and the first, second, and third epitopes are independently selected from CD105, Yb-1, SOX-2, CDH3, or MDM2. In some examples, the composition further comprises third and fourth epitopes, and the first, second, third, and fourth epitopes are independently selected from CD105, Yb-1, SOX-2, CDH3, or MDM2. In some examples, the composition further comprises third, fourth, and fifth epitopes, and the first, second, third, fourth, and fifth epitopes are independently selected from CD105, Yb-1, SOX-2, CDH3, or MDM2.

[0222] In some examples, the composition comprises first and second epitopes independently selected from IGFBP2, HER-2, or IGF-1R. In some examples, the composition further comprises a third epitope, and the first, second, and third epitopes are independently selected from IGFBP2, HER-2, or IGF-1R.

[0223] In some examples, the composition can comprise at least a first epitope of a first antigen, the first epitope being a portion of a peptide derived from HIF-1α. In some examples, the composition can comprise at least a first epitope of a first antigen and at least a second epitope of a second antigen, the first and second epitopes being derived from HIF-1α.

[0224] In some examples, the composition can comprise a nucleotide sequence encoding an amino acid sequence having at least 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the amino acid sequence of DSKTFLSRHSLDMKFSYCDERITELMGYEPEELLGRSIYEYYHALDSDHLTKTHHDMFTKGQVTTGQYRMLAKRGGYVWVETQATVIYN (SEQ ID NO: 82); a nucleotide sequence encoding an amino acid sequence having at least 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the amino acid sequence of SDNVNKYMGLTQFELTGHSVFDFTHP (SEQ ID NO: 83); and a nucleotide sequence encoding an amino acid sequence having at least 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the amino acid sequence of GGYVWVETQATVIYNTKNSQ (SEQ ID NO: 84), and can include a nucleic acid sequence encoding an epitope of the peptide HIF-1α selected from the group consisting of these nucleotide sequences.

[0225] In some examples, the composition can comprise at least a first epitope of a peptide HIF-1α selected from the group consisting of an amino acid sequence having at least 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the amino acid sequence of DSKTFLSRHSLDMKFSYCDERITELMGYEPEELLGRSIYEYYHALDSDHLTKTHHDMFTKGQVTTGQYRMLAKRGGYVWVETQATVIYN (SEQ ID NO: 82); an amino acid sequence having at least 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the amino acid sequence of SDNVNKYMGLTQFELTGHSVFDFTHP (SEQ ID NO: 83); and an amino acid sequence having at least 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the amino acid sequence of GGYVWVETQATVIYNTKNSQ (SEQ ID NO: 84).

[0226] The compositions described herein can contain short epitopes encoded in a single plasmid backbone. In some examples, the plasmid backbone can encode one short epitope. In other examples, the plasmids described herein can encode more than one short epitope. For example, the compositions described herein can encode two short epitopes, three short epitopes, four short epitopes, five short epitopes, six short epitopes, seven short epitopes, eight short epitopes, nine short epitopes, ten short epitopes, eleven short epitopes, twelve short epitopes, thirteen short epitopes, fourteen short epitopes, fifteen short epitopes, sixteen short epitopes, seventeen short epitopes, eighteen short epitopes, nineteen short epitopes, twenty short epitopes or more than twenty short epitopes. In an exemplary example, the plasmid encodes six or fewer short epitopes.

[0227] The compositions described herein can contain extended epitopes encoded in a single plasmid backbone. In some examples, the plasmid can encode one extended epitope. In other examples, the composition can encode more than one extended epitope. For example, the plasmid can encode two extended epitopes, three extended epitopes, four extended epitopes, five extended epitopes, six extended epitopes, seven extended epitopes, eight extended epitopes, nine extended epitopes, ten extended epitopes, eleven extended epitopes, twelve extended epitopes, thirteen extended epitopes, fourteen extended epitopes, fifteen extended epitopes, sixteen extended epitopes, seventeen extended epitopes, eighteen extended epitopes, nineteen extended epitopes, twenty extended epitopes, or more than twenty extended epitopes. In an exemplary example, the plasmid encodes four or fewer extended epitopes.

[0228] The compositions of the breast cancer vaccines described herein can contain short epitopes and extended epitopes in a single plasmid backbone. In some examples, the plasmid can contain one short epitope. In other examples, the compositions of the plasmids described herein can contain more than one short epitope. For example, the compositions of the plasmids described herein can contain two short epitopes, three short epitopes, four short epitopes, five short epitopes, six short epitopes, seven short epitopes, eight short epitopes, nine short epitopes, ten short epitopes, eleven short epitopes, twelve short epitopes, thirteen short epitopes, fourteen short epitopes, fifteen short epitopes, sixteen short epitopes, seventeen short epitopes, eighteen short epitopes, nineteen short epitopes, twenty short epitopes, or more than twenty short epitopes.

[0229] In some examples, the plasmid can encode one extended epitope. In other examples, the compositions described herein can encode more than one extended epitope. For example, the compositions described herein can encode two extended epitopes, three extended epitopes, four extended epitopes, five extended epitopes, six extended epitopes, seven extended epitopes, eight extended epitopes, nine extended epitopes, ten extended epitopes, eleven extended epitopes, twelve extended epitopes, thirteen extended epitopes, fourteen extended epitopes, fifteen extended epitopes, sixteen extended epitopes, seventeen extended epitopes, eighteen extended epitopes nineteen extended epitopes, twenty extended epitopes or more than twenty extended epitopes.

[0230] Plasmids for compositions containing more than one sequence encoding an epitope can contain a spacer between each epitope sequence. In some examples, the sequences of short epitopes can be encoded tandemly without using a spacer. In some examples, the sequences of extended epitopes can be encoded tandemly without using a spacer. In some examples, the sequences of short epitopes can be encoded tandemly using a spacer. In some examples, the sequences of extended epitopes can be encoded tandemly using a spacer.

[0231] In an exemplary example, the composition can be a plasmid-based vaccine containing short and extended antigenic epitopes. The plasmid(s) of the vaccine can be constructed using a 4 kB plasmid backbone (e.g., pUMVC3 or pNGVL3). In many cases, the plasmid can contain an antibiotic resistance gene. For example, pUMVC3 contains a kanamycin resistance gene in addition to an origin of replication for selection and propagation in bacteria. In some examples, the multiple cloning site in pUMVC3 can be adjacent to eukaryotic transcriptional control elements for promoting the expression of the inserted sequence (e.g., gene cassette) in eukaryotic cells. For example, the inserted sequence can be an epitope.

[0232] In an exemplary example, the nucleic acid coding sequence of the antigenic epitope peptide can be assembled with a Kozak consensus translation initiation sequence, a stop codon, and a cloning site in the plasmid backbone. Standard molecular techniques known to those skilled in the art, including synthetic oligonucleotides, polymerase chain reaction amplification, restriction endonucleases, and nucleic acid ligases (e.g., DNA ligase), can be used to generate nucleic acids (e.g., DNA fragments) and insert the nucleic acid fragments into the plasmid vector backbone.

[0233] In some examples, the plasmid can contain a nucleic acid sequence encoding at least one tag. In some examples, the tag can be translated into a peptide. Any nucleic acid sequence for tags known to those skilled in the art can be used with the plasmids described herein. For example, the tag can be a histidine tag with 3 histidine residues, a histidine tag with 4 histidine residues, a histidine tag with 5 histidine residues, a histidine tag with 6 histidine residues, or others. The expression of the tag in a subject can be determined using any suitable technique known to those skilled in the art.

[0234] In some examples, the plasmid can be sequenced using any sequencing technique known to those skilled in the art such that the results of the sequencing technique provide nucleotide-level resolution of the entire plasmid.

[0235] In some aspects, the composition can be a multi-antigen breast cancer vaccine. For example, the multi-antigen breast cancer vaccine can contain multiple antigens. In some examples, the expression of one antigen can affect the expression of a different antigen. In some examples, the expression of more than one antigen can affect the expression of a different antigen. In some examples, the expression of one antigen can affect the expression of more than one different antigen. In some examples, the expression of one antigen may not affect the expression of a different antigen. In some examples, the expression of more than one antigen may not affect the expression of a different antigen. In some examples, the expression of one antigen may not affect the expression of more than one different antigen. For example, antigenic competition can limit the immunogenicity of the multi-antigen vaccine. Using any technique known to those skilled in the art, it can be determined whether the immune response induced after administration of the multi-antigen vaccine is of a magnitude comparable to that of each antigen as a single antigen vaccine. For example, ELISPOT (e.g., for the secretion of IFNγ) can determine the magnitude of the immune response. In some examples, ELISPOT can detect rodent, non-human primate, or human peptides. Plasmids - Survivin, HIF-1A, IGF-1R and / or IGFBP2

[0236] The compositions described herein can include a nucleic acid-based vaccine comprising a plasmid encoding one or more epitopes selected from survivin, HIF-1A, IGF-1R, or IGFBP2. Optionally, the epitope can be derived from a human protein, and the nucleic acid sequence encoding the epitope can be incorporated into a nucleic acid construct designed to induce expression of the epitope in a subject after administration. For example, the encoded epitope from the nucleic acid construct can enable an immune response against at least one epitope to be tuned, amplified, attenuated, suppressed, or eliminated for a specific set of proteins (e.g., self-proteins).

[0237] In some cases, the vaccines described herein are peptide-based vaccines. Peptide-based vaccines can include a plasmid encoding one or more epitopes selected from survivin, HIF-1A, IGF-1R, or IGFBP2. The epitope can be derived from a human protein that can be used directly in the peptide-based vaccine.

[0238] In some examples, the peptide or nucleic acid construct can be optimized for protein- or plasmid-based vaccination to induce, amplify, or tune a TH1 immune response. In some examples, the epitope can be an extended TH1 epitope. In other examples, the peptide or nucleic acid construct can be optimized for protein- or plasmid-based vaccination to suppress, attenuate, or eliminate a pathological response in a subject (e.g., human or animal) in need thereof.

[0239] The compositions described herein can include a plasmid containing a nucleic acid sequence for expressing at least one epitope in a subject after administration of the composition (e.g., vaccine).

[0240] Any plasmid known to those of ordinary skill in the art suitable for pharmaceutical use for the expression of a nucleic acid sequence The mid-skeleton (e.g., vector) can be used in the compositions described herein.

[0241] The vector can be a circular plasmid or a linear nucleic acid. The circular plasmid or linear nucleic acid can be capable of directing the expression of a specific nucleotide sequence in a suitable target cell. The vector can have a promoter operably linked to a nucleotide sequence encoding a polypeptide that can be operably linked to a termination signal. The vector can also contain sequences required for proper translation of the nucleotide sequence. A vector containing a nucleotide sequence of interest can be chimeric, which means that at least one of its components is heterologous to at least one of the other components. The expression of the nucleotide sequence in the expression cassette can be under the control of a constitutive promoter or an inducible promoter that can initiate transcription only when the host cell is exposed to a specific external stimulus.

[0242] The vector can be a plasmid. The plasmid can be useful for transfection of cells with nucleic acid encoding a polypeptide, and the transformed host cells can be cultured and maintained under conditions in which expression of the polypeptide occurs.

[0243] The plasmid can contain a nucleic acid sequence encoding one or more of the various polypeptides disclosed herein. A single plasmid can contain a coding sequence for a single polypeptide or a coding sequence for more than one polypeptide. Optionally, the plasmid can further contain a coding sequence encoding an adjuvant such as an immunostimulatory molecule such as a cytokine.

[0244] The plasmid can further include a start codon that can be upstream of the coding sequence and a stop codon that can be downstream of the coding sequence. The start and stop codons can be in-frame with the coding sequence. The plasmid can also include a promoter operably linked to the coding sequence and an enhancer upstream of the coding sequence. The enhancer can be a human actin, human myosin, human hemoglobin, human muscle creatine or a viral enhancer such as those derived from CMV, FMDV, RSV or EBV. Polynucleotide function enhancement is described in U.S. Patent Nos. 5,593,972, 5,962,428 and W094 / 016737.

[0245] To maintain the plasmid episomally and produce multiple copies of the plasmid in the cell, the plasmid can also include a mammalian origin of replication. The plasmid can be pVAXI, pCEP4 or pREP4 manufactured by Invitrogen (San Diego, CA).

[0246] The plasmid can include regulatory sequences that can be well-suited for gene expression in the cells to which the plasmid is administered. The coding sequence can include codons that can enable more efficient transcription of the coding sequence in the host cell.

[0247] In some examples, commercially available plasmid backbones can be used. For example, the plasmid pUMVC3 can be used. In some examples, the commercially available plasmid backbones can be modified, mutated, genetically engineered or cloned before use. In other examples, non-commercially available plasmid backbones can be used.

[0248] Additional plasmids can include pSE420 (Invitrogen, San Diego, Calif), which can be used for protein production in Escherichia coli (E. coli). The plasmid can also be pYES2 (Invitrogen, San Diego, Calif), which can be used for protein production in the Saccharomyces cerevisiae strain of yeast. The plasmid can also be from the MAXBAC™ Complete Baculovirus Expression System (Invitrogen, San Diego, Calif), which can be used for protein production in insect cells. The plasmid can also be pcDNA I or pcDNA3 (Invitrogen, San Diego, Calif), which can be used for protein production in mammalian cells such as Chinese hamster ovary (CHO) cells.

[0249] The vector can be a circular plasmid that can transform target cells by integration into the cell genome or can exist extrachromosomally (e.g., an autonomously replicating plasmid having an origin of replication). Exemplary vectors include pVAX, pcDNA3.0 or provax, or any other expression vector capable of expressing DNA encoding an antigen and causing the cell to translate the sequence into an antigen recognized by the immune system.

[0250] Nucleic acid-based vaccines can also be linear nucleic acid vaccines or linear expression cassettes ("LECs") that can be efficiently delivered to a subject by electroporation to express one or more polypeptides disclosed herein. An LEC can be any linear DNA lacking any phosphate backbone. The DNA can encode one or more polypeptides disclosed herein. The LEC can contain a promoter, intron, stop codon, and / or polyadenylation signal. Expression of the polypeptide can be controlled by the promoter. The LEC cannot contain any antibiotic resistance gene and / or phosphate backbone. The LEC cannot contain other nucleic acid sequences unrelated to polypeptide expression.

[0251] The LEC can be derived from any plasmid that can be linearized. The plasmid can express a polypeptide. Exemplary plasmids include pNP (Puerto Rico / 34), pM2 (New Caledonia / 99), WLV009, pVAX, pcDNA3.0, provax, or any other expression vector capable of expressing DNA encoding an antigen and translating the sequence into an antigen recognized by the immune system in a cell.

[0252] Before inserting the nucleic acid sequence of at least one epitope, the plasmid backbone can be less than about 500 bp, about 1.0 kB, about 1.2 kB, about 1.4 kB, about 1.6 kB, about 1.8 kB, about 2.0 kB, about 2.2 kB, about 2.4 kB, about 2.6 kB, about 2.8 kB, about 3.0 kB, about 3.2 kB, about 3.4 kB, about 3.6 kB, about 3.8 kB, about 4.0 kB, about 4.2 kB, about 4.4 kB, about 4.6 kB, about 4.8 kB, about 5.0 kB, about 5.2 kB, about 5.4 kB, about 5.6 kB, about 5.8 kB, about 6.0 kB, about 6.2 kB, about 6.4 kB, about 6.6 kB, about 6.8 kB, about 7.0 kB, about 7.2 kB, about 7.4 kB, about 7.6 kB, about 7.8 kB, about 8.0 kB, about 8.2 kB, about 8.4 kB, about 8.6 kB, about 8.8 kB, about 9.0 kB, about 9.2 kB, about 9.4 kB, about 9.6 kB, about 9.8 kB, about 10.0 kB, about 10.2 kB, about 10.4 kB, about 10.6 kB, about 10.8 kB, about 11.0 kB, about 11.2 kB, about 11.4 kB, about 11.6 kB, about 11.8 kB, about 12.0 kB, about 12.2 kB, about 12.4 kB, about 12.6 kB, about 12.8 kB, about 13.0 kB, about 13.2 kB, about 13.4 kB, about 13.6 kB, about 13.8 kB, about 14 kB, about 14.5 kB, about 15 kB, about 15.5 kB, about 16 kB, about 16.5 kB, about 17 kB, about 17.5 kB, about 18 kB, about 18.5 kB, about 19 kB, about 19.5 kB, about 20 kB, about 30 kB, about 40 kB, about 50 kB, about 60 kB, about 70 kB, about 80 kB, about 90 kB, about 100 kB, about 110 kB, about 120 kB, about 130 kB, about 140 kB, about 150 kB, about 160 kB, about 170 kB, about 180 kB, about 190 kB or about 200 kB in length. In an exemplary example, before the addition of the nucleic acid sequence encoding at least one epitope, the plasmid is about 4 kB in length.

[0253] In some examples, the compositions described herein can include one plasmid. In other examples, the compositions described herein can include more than one plasmid. For example, the compositions described herein can include two plasmids, three plasmids, four plasmids, five plasmids, six plasmids, seven plasmids, eight plasmids, nine plasmids, ten plasmids, eleven plasmids, twelve plasmids, thirteen plasmids, fourteen plasmids, fifteen plasmids, sixteen plasmids, seventeen plasmids, eighteen plasmids, nineteen plasmids, twenty plasmids, or more than twenty plasmids.

[0254] The nucleic acid encoding at least one epitope of the plasmid can be from any species such that the epitope expressed from the nucleic acid elicits an immune response in a subject. In some examples, the subject can be a rodent, non-human primate, or human. The nucleic acid encoding the epitope of the plasmid can be isolated from any nucleic acid source using methods and techniques known to those of skill in the art. The nucleic acid encoding the epitope of the plasmid can be cloned into the plasmid backbone using methods and techniques known to those of skill in the art.

[0255] In some examples, the nucleic acid sequence encoding the epitope can be an endogenous nucleic acid sequence to the subject. For example, the nucleic acid sequence of survivin from human can be used to express survivin in humans. In other examples, the nucleic acid sequence of the antigenic epitope can be an exogenous nucleic acid sequence to the subject. For example, the nucleic acid sequence of survivin from non-human can be used to express survivin in humans.

[0256] In some examples, the nucleic acid sequence for expressing an antigenic epitope can be a wild-type nucleic acid sequence. For example, the nucleic acid sequence of natural origin of survivin in a certain genome can be used to express survivin in a subject. In other examples, the nucleic acid sequence encoding the epitope can be a synthetic nucleic acid sequence. For example, the nucleic acid sequence of natural origin of survivin in a certain genome can be modified using molecular techniques known to those skilled in the art and used for the expression of survivin in a subject.

[0257] In some examples, the nucleic acid sequence encoding the epitope can be a nucleic acid sequence endogenous to the subject. For example, the nucleic acid sequence of HIF-1A derived from humans can be used to express HIF-1A in humans. In other examples, the nucleic acid sequence of the antigenic epitope can be a nucleic acid sequence exogenous to the subject. For example, the nucleic acid sequence of HIF-1A derived from non-humans can be used to express HIF-1A in humans.

[0258] In some examples, the nucleic acid sequence for expressing an antigenic epitope can be a wild-type nucleic acid sequence. For example, the nucleic acid sequence of natural origin of HIF-1A in a certain genome can be used to express HIF-1A in a subject. In other examples, the nucleic acid sequence encoding the epitope can be a synthetic nucleic acid sequence. For example, the nucleic acid sequence of natural origin of HIF-1A in a certain genome can be modified using molecular techniques known to those skilled in the art and used for the expression of HIF-1A in a subject.

[0259] In some examples, the nucleic acid sequence encoding the epitope can be a nucleic acid sequence endogenous to the subject. For example, the nucleic acid sequence of IGFBP2 derived from humans can be used to express IGFBP2 in humans. In other examples, the nucleic acid sequence of the antigenic epitope can be a nucleic acid sequence exogenous to the subject. For example, the nucleic acid sequence of IGFBP2 derived from non-humans can be used to express IGFBP2 in humans.

[0260] In some examples, the nucleic acid sequence for expressing an antigenic epitope can be a wild-type nucleic acid sequence. For example, the nucleic acid sequence of natural origin of IGFBP2 in a certain genome can be used to express IGFBP2 in a subject. In other examples, the nucleic acid sequence encoding an epitope can be a synthetic nucleic acid sequence. For example, the nucleic acid sequence of natural origin of IGFBP2 in a certain genome can be modified using molecular techniques known to those skilled in the art and used for the expression of IGFBP2 in a subject.

[0261] In some examples, the nucleic acid sequence encoding an epitope can be a nucleic acid sequence endogenous to the subject. For example, the nucleic acid sequence of IGF-1R derived from humans can be used to express IGF-1R in humans. In other examples, the nucleic acid sequence of an antigenic epitope can be a nucleic acid sequence exogenous to the subject. For example, the nucleic acid sequence of IGF-1R derived from non-humans can be used to express IGF-1R in humans.

[0262] In some examples, the nucleic acid sequence for expressing an antigenic epitope can be a wild-type nucleic acid sequence. For example, the nucleic acid sequence of natural origin of IGF-1R in a certain genome can be used to express IGF-1R in a subject. In other examples, the nucleic acid sequence encoding an epitope can be a synthetic nucleic acid sequence. For example, the nucleic acid sequence of natural origin of IGF-1R in a certain genome can be modified using molecular techniques known to those skilled in the art and used for the expression of IGF-1R in a subject.

[0263] In some examples, the composition can include a nucleic acid encoding one or more epitopes derived from a protein, survivin, HIF-1A, IGFBP2, and IGF-1R. In some cases, the plasmid can include a nucleic acid sequence encoding a polypeptide that includes at least 70% sequence identity to IGFBP-2 (SEQ ID NO: 54). In some cases, the plasmid can include a nucleic acid sequence encoding a polypeptide that includes at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to IGFBP-2 (SEQ ID NO: 54). In some examples, the plasmid can include a nucleic acid sequence encoding a polypeptide that includes at least 80% sequence identity to IGFBP-2 (SEQ ID NO: 54). In some examples, the plasmid can include a nucleic acid sequence encoding a polypeptide that includes at least 90% sequence identity to IGFBP-2 (SEQ ID NO: 54). Optionally, the plasmid can include a nucleic acid sequence encoding a polypeptide that includes at least 95% sequence identity to IGFBP-2 (SEQ ID NO: 54). Optionally, the plasmid can include a nucleic acid sequence encoding a polypeptide that includes at least 99% sequence identity to IGFBP-2 (SEQ ID NO: 54). In some cases, the plasmid can include a nucleic acid sequence encoding a polypeptide that includes 100% sequence identity to IGFBP-2 (SEQ ID NO: 54). In some cases, the plasmid can include a nucleic acid sequence encoding a polypeptide that consists of 100% sequence identity to IGFBP-2 (SEQ ID NO: 54).

[0264] In some examples, the plasmid can include a nucleic acid sequence encoding a polypeptide that includes at least 70% sequence identity to at least 100 to at least 163 contiguous amino acids of IGFBP-2 (SEQ ID NO: 54). In some examples, the plasmid can include a nucleic acid sequence encoding a polypeptide that includes at least 70% sequence identity to at least 105 to at least 160, at least 110 to at least 155, or at least 120 to at least 145 contiguous amino acids of IGFBP-2 (SEQ ID NO: 54).

[0265] In some examples, the plasmid can include a nucleic acid sequence encoding a polypeptide that includes at least 80% sequence identity to at least 100 to at least 163 contiguous amino acids of IGFBP-2 (SEQ ID NO: 54). In some examples, the plasmid can include a nucleic acid sequence encoding a polypeptide that includes at least 80% sequence identity to at least 105 to at least 160, at least 110 to at least 155, or at least 120 to at least 145 contiguous amino acids of IGFBP-2 (SEQ ID NO: 54).

[0266] In some examples, the plasmid can include a nucleic acid sequence encoding a polypeptide that includes at least 90% sequence identity to at least 100 to at least 163 contiguous amino acids of IGFBP-2 (SEQ ID NO: 54). In some examples, the plasmid can include a nucleic acid sequence encoding a polypeptide that includes at least 90% sequence identity to at least 105 to at least 160, at least 110 to at least 155, or at least 120 to at least 145 contiguous amino acids of IGFBP-2 (SEQ ID NO: 54).

[0267] In some examples, the plasmid can include a nucleic acid sequence encoding a polypeptide that includes at least 95% sequence identity to at least 100 to at least 163 contiguous amino acids of IGFBP-2 (SEQ ID NO: 54). In some examples, the plasmid can include a nucleic acid sequence encoding a polypeptide that includes at least 95% sequence identity to at least 105 to at least 160, at least 110 to at least 155, or at least 120 to at least 145 contiguous amino acids of IGFBP-2 (SEQ ID NO: 54).

[0268] In some examples, the plasmid can include a nucleic acid sequence encoding a polypeptide having 100% sequence identity to at least 100 to at least 163 contiguous amino acids of IGFBP-2 (SEQ ID NO: 54). In some examples, the plasmid can include a nucleic acid sequence encoding a polypeptide having 100% sequence identity to at least 105 to at least 160, at least 110 to at least 155, or at least 120 to at least 145 contiguous amino acids of IGFBP-2 (SEQ ID NO: 54).

[0269] In some examples, the plasmid can include a nucleic acid sequence encoding a polypeptide consisting of 100% sequence identity to at least 100 to at least 163 contiguous amino acids of IGFBP-2 (SEQ ID NO: 54). In some examples, the plasmid can include a nucleic acid sequence encoding a polypeptide consisting of 100% sequence identity to at least 105 to at least 160, at least 110 to at least 155, or at least 120 to at least 145 contiguous amino acids of IGFBP-2 (SEQ ID NO: 54).

[0270] In some cases, the plasmid can comprise a nucleic acid sequence having at least 50% sequence identity to IGFBP-2 (SEQ ID NO: 43). In some cases, the plasmid can comprise a nucleic acid sequence having at least 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to IGFBP-2 (SEQ ID NO: 43). In some examples, the plasmid can comprise a nucleic acid sequence having at least 70% sequence identity to IGFBP-2 (SEQ ID NO: 43). In some examples, the plasmid can comprise a nucleic acid sequence having at least 80% sequence identity to IGFBP-2 (SEQ ID NO: 43). In some examples, the plasmid can comprise a nucleic acid sequence having at least 90% sequence identity to IGFBP-2 (SEQ ID NO: 43). Optionally, the plasmid can comprise a nucleic acid sequence having at least 95% sequence identity to IGFBP-2 (SEQ ID NO: 43). Optionally, the plasmid can comprise a nucleic acid sequence having at least 99% sequence identity to IGFBP-2 (SEQ ID NO: 43). In some cases, the plasmid can comprise a nucleic acid sequence having 100% sequence identity to IGFBP-2 (SEQ ID NO: 43). In some cases, the plasmid can comprise a nucleic acid sequence consisting of 100% sequence identity to IGFBP-2 (SEQ ID NO: 43).

[0271] In some cases, the plasmid can contain a nucleic acid sequence encoding a polypeptide having at least 70% sequence identity to survivin (SEQ ID NO: 85). In some cases, the plasmid can contain a nucleic acid sequence encoding a polypeptide having at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to survivin (SEQ ID NO: 85). In some examples, the plasmid can contain a nucleic acid sequence encoding a polypeptide having at least 80% sequence identity to survivin (SEQ ID NO: 85). In some examples, the plasmid can contain a nucleic acid sequence encoding a polypeptide having at least 90% sequence identity to survivin (SEQ ID NO: 85). Optionally, the plasmid can contain a nucleic acid sequence encoding a polypeptide having at least 95% sequence identity to survivin (SEQ ID NO: 85). Optionally, the plasmid can contain a nucleic acid sequence encoding a polypeptide having at least 99% sequence identity to survivin (SEQ ID NO: 85). In some cases, the plasmid can contain a nucleic acid sequence encoding a polypeptide having 100% sequence identity to survivin (SEQ ID NO: 85). In some cases, the plasmid can contain a nucleic acid sequence encoding a polypeptide consisting of 100% sequence identity to survivin (SEQ ID NO: 85).

[0272] In some examples, the plasmid can comprise a nucleic acid sequence encoding a polypeptide having at least 70% sequence identity to at least 10 to at least 38 contiguous amino acids of survivin (SEQ ID NO: 85). In some examples, the plasmid can comprise a nucleic acid sequence encoding a polypeptide having at least 70% sequence identity to at least 12 to at least 35, at least 15 to at least 30 or at least 20 to at least 25 contiguous amino acids of survivin (SEQ ID NO: 85). In some examples, the plasmid can comprise a nucleic acid sequence encoding a polypeptide having at least 70% sequence identity to at least 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 32, 34 or 36 contiguous amino acids of survivin (SEQ ID NO: 85).

[0273] In some examples, the plasmid can comprise a nucleic acid sequence encoding a polypeptide having at least 80% sequence identity to at least 10 to at least 38 contiguous amino acids of survivin (SEQ ID NO: 85). In some examples, the plasmid can comprise a nucleic acid sequence encoding a polypeptide having at least 80% sequence identity to at least 12 to at least 35, at least 15 to at least 30 or at least 20 to at least 25 contiguous amino acids of survivin (SEQ ID NO: 85). In some examples, the plasmid can comprise a nucleic acid sequence encoding a polypeptide having at least 80% sequence identity to at least 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 32, 34 or 36 contiguous amino acids of survivin (SEQ ID NO: 85).

[0274] In some examples, the plasmid can include a nucleic acid sequence encoding a polypeptide having at least 90% sequence identity to at least 10 to at least 38 contiguous amino acids of survivin (SEQ ID NO: 85). In some examples, the plasmid can include a nucleic acid sequence encoding a polypeptide having at least 90% sequence identity to at least 12 to at least 35, at least 15 to at least 30, or at least 20 to at least 25 contiguous amino acids of survivin (SEQ ID NO: 85). In some examples, the plasmid can include a nucleic acid sequence encoding a polypeptide having at least 90% sequence identity to at least 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 32, 34, or 36 contiguous amino acids of survivin (SEQ ID NO: 85).

[0275] In some examples, the plasmid can include a nucleic acid sequence encoding a polypeptide having at least 95% sequence identity to at least 10 to at least 38 contiguous amino acids of survivin (SEQ ID NO: 85). In some examples, the plasmid can include a nucleic acid sequence encoding a polypeptide having at least 95% sequence identity to at least 12 to at least 35, at least 15 to at least 30, or at least 20 to at least 25 contiguous amino acids of survivin (SEQ ID NO: 85). In some examples, the plasmid can include a nucleic acid sequence encoding a polypeptide having at least 95% sequence identity to at least 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 32, 34, or 36 contiguous amino acids of survivin (SEQ ID NO: 85).

[0276] In some examples, the plasmid can include a nucleic acid sequence encoding a polypeptide having 100% sequence identity to at least 10 to at least 38 contiguous amino acids of survivin (SEQ ID NO: 85). In some examples, the plasmid can include a nucleic acid sequence encoding a polypeptide having 100% sequence identity to at least 12 to at least 35, at least 15 to at least 30, or at least 20 to at least 25 contiguous amino acids of survivin (SEQ ID NO: 85). In some examples, the plasmid can include a nucleic acid sequence encoding a polypeptide having 100% sequence identity to at least 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 32, 34, or 36 contiguous amino acids of survivin (SEQ ID NO: 85).

[0277] In some examples, the plasmid can include a nucleic acid sequence encoding a polypeptide consisting of 100% sequence identity to at least 10 to at least 38 contiguous amino acids of survivin (SEQ ID NO: 85). In some examples, the plasmid can include a nucleic acid sequence encoding a polypeptide consisting of 100% sequence identity to at least 12 to at least 35, at least 15 to at least 30, or at least 20 to at least 25 contiguous amino acids of survivin (SEQ ID NO: 85). In some examples, the plasmid can include a nucleic acid sequence encoding a polypeptide consisting of 100% sequence identity to at least 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 32, 34, or 36 contiguous amino acids of survivin (SEQ ID NO: 85).

[0278] In some cases, the plasmid can comprise a nucleic acid sequence having at least 50% sequence identity to survivin (SEQ ID NO: 86). In some cases, the plasmid can comprise a nucleic acid sequence having at least 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to survivin (SEQ ID NO: 86). In some examples, the plasmid can comprise a nucleic acid sequence having at least 70% sequence identity to survivin (SEQ ID NO: 86). In some examples, the plasmid can comprise a nucleic acid sequence having at least 80% sequence identity to survivin (SEQ ID NO: 86). In some examples, the plasmid can comprise a nucleic acid sequence having at least 90% sequence identity to survivin (SEQ ID NO: 86). Optionally, the plasmid can comprise a nucleic acid sequence having at least 95% sequence identity to survivin (SEQ ID NO: 86). Optionally, the plasmid can comprise a nucleic acid sequence having at least 99% sequence identity to survivin (SEQ ID NO: 86). In some cases, the plasmid can comprise a nucleic acid sequence having 100% sequence identity to survivin (SEQ ID NO: 86). In some cases, the plasmid can comprise a nucleic acid sequence consisting of 100% sequence identity to survivin (SEQ ID NO: 86).

[0279] In some cases, the plasmid can include a nucleic acid sequence encoding a polypeptide having at least 70% sequence identity to HIF-1A (SEQ ID NO: 87). In some cases, the plasmid can include a nucleic acid sequence encoding a polypeptide having at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to HIF-1A (SEQ ID NO: 87). In some examples, the plasmid can include a nucleic acid sequence encoding a polypeptide having at least 80% sequence identity to HIF-1A (SEQ ID NO: 87). In some examples, the plasmid can include a nucleic acid sequence encoding a polypeptide having at least 90% sequence identity to HIF-1A (SEQ ID NO: 87). Optionally, the plasmid can include a nucleic acid sequence encoding a polypeptide having at least 95% sequence identity to HIF-1A (SEQ ID NO: 87). Optionally, the plasmid can include a nucleic acid sequence encoding a polypeptide having at least 99% sequence identity to HIF-1A (SEQ ID NO: 87). In some cases, the plasmid can include a nucleic acid sequence encoding a polypeptide having 100% sequence identity to HIF-1A (SEQ ID NO: 87). In some cases, the plasmid can include a nucleic acid sequence encoding a polypeptide consisting of 100% sequence identity to HIF-1A (SEQ ID NO: 87).

[0280] In some examples, the plasmid can include a nucleic acid sequence encoding a polypeptide having at least 70% sequence identity to at least 40 to at least 89 contiguous amino acids of HIF-1A (SEQ ID NO: 87). In some examples, the plasmid can include a nucleic acid sequence encoding a polypeptide having at least 70% sequence identity to at least 40 to at least 85, at least 50 to at least 80, at least 55 to at least 75, or at least 60 to at least 70 contiguous amino acids of HIF-1A (SEQ ID NO: 87). In some examples, the plasmid can include a nucleic acid sequence encoding a polypeptide having at least 70% sequence identity to at least 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, or 88 contiguous amino acids of HIF-1A (SEQ ID NO: 87).

[0281] In some examples, the plasmid can include a nucleic acid sequence encoding a polypeptide having at least 80% sequence identity to at least 40 to at least 89 contiguous amino acids of HIF-1A (SEQ ID NO: 87). In some examples, the plasmid can include a nucleic acid sequence encoding a polypeptide having at least 80% sequence identity to at least 40 to at least 85, at least 50 to at least 80, at least 55 to at least 75, or at least 60 to at least 70 contiguous amino acids of HIF-1A (SEQ ID NO: 87). In some examples, the plasmid can include a nucleic acid sequence encoding a polypeptide having at least 80% sequence identity to at least 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, or 88 contiguous amino acids of HIF-1A (SEQ ID NO: 87).

[0282] In some examples, the plasmid can include a nucleic acid sequence encoding a polypeptide having at least 90% sequence identity to at least 40 to at least 89 contiguous amino acids of HIF-1A (SEQ ID NO: 87). In some examples, the plasmid can include a nucleic acid sequence encoding a polypeptide having at least 90% sequence identity to at least 40 to at least 85, at least 50 to at least 80, at least 55 to at least 75, or at least 60 to at least 70 contiguous amino acids of HIF-1A (SEQ ID NO: 87). In some examples, the plasmid can include a nucleic acid sequence encoding a polypeptide having at least 90% sequence identity to at least 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, or 88 contiguous amino acids of HIF-1A (SEQ ID NO: 87).

[0283] In some examples, the plasmid can include a nucleic acid sequence encoding a polypeptide having at least 95% sequence identity to at least 40 to at least 89 contiguous amino acids of HIF-1A (SEQ ID NO: 87). In some examples, the plasmid can include a nucleic acid sequence encoding a polypeptide having at least 95% sequence identity to at least 40 to at least 85, at least 50 to at least 80, at least 55 to at least 75, or at least 60 to at least 70 contiguous amino acids of HIF-1A (SEQ ID NO: 87). In some examples, the plasmid can include a nucleic acid sequence encoding a polypeptide having at least 95% sequence identity to at least 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, or 88 contiguous amino acids of HIF-1A (SEQ ID NO: 87).

[0284] In some examples, the plasmid can include a nucleic acid sequence encoding a polypeptide having 100% sequence identity to at least 40 to at least 89 contiguous amino acids of HIF-1A (SEQ ID NO: 87). In some examples, the plasmid can include a nucleic acid sequence encoding a polypeptide having 100% sequence identity to at least 40 to at least 85, at least 50 to at least 80, at least 55 to at least 75, or at least 60 to at least 70 contiguous amino acids of HIF-1A (SEQ ID NO: 87). In some examples, the plasmid can include a nucleic acid sequence encoding a polypeptide having 100% sequence identity to at least 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, or 88 contiguous amino acids of HIF-1A (SEQ ID NO: 87).

[0285] In some examples, the plasmid can include a nucleic acid sequence encoding a polypeptide consisting of 100% sequence identity to at least 40 to at least 89 contiguous amino acids of HIF-1A (SEQ ID NO: 87). In some examples, the plasmid can include a nucleic acid sequence encoding a polypeptide consisting of 100% sequence identity to at least 40 to at least 85, at least 50 to at least 80, at least 55 to at least 75, or at least 60 to at least 70 contiguous amino acids of HIF-1A (SEQ ID NO: 87). In some examples, the plasmid can include a nucleic acid sequence encoding a polypeptide consisting of 100% sequence identity to at least 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, or 88 contiguous amino acids of HIF-1A (SEQ ID NO: 87).

[0286] In some cases, the plasmid can comprise a nucleic acid sequence having at least 50% sequence identity to HIF-1A (SEQ ID NO: 88). In some cases, the plasmid can comprise a nucleic acid sequence having at least 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to HIF-1A (SEQ ID NO: 88). In some examples, the plasmid can comprise a nucleic acid sequence having at least 70% sequence identity to HIF-1A (SEQ ID NO: 88). In some examples, the plasmid can comprise a nucleic acid sequence having at least 80% sequence identity to HIF-1A (SEQ ID NO: 88). In some examples, the plasmid can comprise a nucleic acid sequence having at least 90% sequence identity to HIF-1A (SEQ ID NO: 88). Optionally, the plasmid can comprise a nucleic acid sequence having at least 95% sequence identity to HIF-1A (SEQ ID NO: 88). Optionally, the plasmid can comprise a nucleic acid sequence having at least 99% sequence identity to HIF-1A (SEQ ID NO: 88). In some cases, the plasmid can comprise a nucleic acid sequence having 100% sequence identity to HIF-1A (SEQ ID NO: 88). In some cases, the plasmid can comprise a nucleic acid sequence consisting of 100% sequence identity to HIF-1A (SEQ ID NO: 88).

[0287] In some cases, the plasmid can contain a nucleic acid sequence encoding a polypeptide that contains at least 70% sequence identity to IGF-IR (SEQ ID NO: 73). In some cases, the plasmid can contain a nucleic acid sequence encoding a polypeptide that contains at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to IGF-IR (SEQ ID NO: 73). In some examples, the plasmid can contain a nucleic acid sequence encoding a polypeptide that contains at least 80% sequence identity to IGF-IR (SEQ ID NO: 73). In some examples, the plasmid can contain a nucleic acid sequence encoding a polypeptide that contains at least 90% sequence identity to IGF-IR (SEQ ID NO: 73). Optionally, the plasmid can contain a nucleic acid sequence encoding a polypeptide that contains at least 95% sequence identity to IGF-IR (SEQ ID NO: 73). Optionally, the plasmid can contain a nucleic acid sequence encoding a polypeptide that contains at least 99% sequence identity to IGF-IR (SEQ ID NO: 73). In some cases, the plasmid can contain a nucleic acid sequence encoding a polypeptide that contains 100% sequence identity to IGF-IR (SEQ ID NO: 73). In some cases, the plasmid can contain a nucleic acid sequence encoding a polypeptide that consists of 100% sequence identity to IGF-IR (SEQ ID NO: 73).

[0288] In some examples, the plasmid can contain a nucleic acid sequence encoding a polypeptide that contains at least 70% sequence identity to at least 50 to at least 104 contiguous amino acids of IGF-1R (SEQ ID NO: 73). In some examples, the plasmid can contain a nucleic acid sequence encoding a polypeptide that contains at least 70% sequence identity to at least 55 to at least 100, at least 60 to at least 90 or at least 70 to at least 80 contiguous amino acids of IGF-1R (SEQ ID NO: 73).

[0289] In some examples, the plasmid can include a nucleic acid sequence encoding a polypeptide that includes at least 80% sequence identity to at least 50 to at least 104 contiguous amino acids of IGF-1R (SEQ ID NO: 73). In some examples, the plasmid can include a nucleic acid sequence encoding a polypeptide that includes at least 80% sequence identity to at least 55 to at least 100, at least 60 to at least 90, or at least 70 to at least 80 contiguous amino acids of IGF-1R (SEQ ID NO: 73).

[0290] In some examples, the plasmid can include a nucleic acid sequence encoding a polypeptide that includes at least 90% sequence identity to at least 50 to at least 104 contiguous amino acids of IGF-1R (SEQ ID NO: 73). In some examples, the plasmid can include a nucleic acid sequence encoding a polypeptide that includes at least 90% sequence identity to at least 55 to at least 100, at least 60 to at least 90, or at least 70 to at least 80 contiguous amino acids of IGF-1R (SEQ ID NO: 73).

[0291] In some examples, the plasmid can include a nucleic acid sequence encoding a polypeptide that includes at least 95% sequence identity to at least 50 to at least 104 contiguous amino acids of IGF-1R (SEQ ID NO: 73). In some examples, the plasmid can include a nucleic acid sequence encoding a polypeptide that includes at least 95% sequence identity to at least 55 to at least 100, at least 60 to at least 90, or at least 70 to at least 80 contiguous amino acids of IGF-1R (SEQ ID NO: 73).

[0292] In some examples, the plasmid can include a nucleic acid sequence encoding a polypeptide having 100% sequence identity to at least 50 to at least 104 contiguous amino acids of IGF-1R (SEQ ID NO: 73). In some examples, the plasmid can include a nucleic acid sequence encoding a polypeptide having 100% sequence identity to at least 55 to at least 100, at least 60 to at least 90, or at least 70 to at least 80 contiguous amino acids of IGF-1R (SEQ ID NO: 73).

[0293] In some examples, the plasmid can include a nucleic acid sequence encoding a polypeptide consisting of 100% sequence identity to at least 50 to at least 104 contiguous amino acids of IGF-1R (SEQ ID NO: 73). In some examples, the plasmid can include a nucleic acid sequence encoding a polypeptide consisting of 100% sequence identity to at least 55 to at least 100, at least 60 to at least 90, or at least 70 to at least 80 contiguous amino acids of IGF-1R (SEQ ID NO: 73).

[0294] In some cases, the plasmid can comprise a nucleic acid sequence having at least 50% sequence identity to IGF-IR (SEQ ID NO: 63). In some cases, the plasmid can comprise a nucleic acid sequence having at least 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to IGF-IR (SEQ ID NO: 63). In some examples, the plasmid can comprise a nucleic acid sequence having at least 70% sequence identity to IGF-IR (SEQ ID NO: 63). In some examples, the plasmid can comprise a nucleic acid sequence having at least 80% sequence identity to IGF-IR (SEQ ID NO: 63). In some examples, the plasmid can comprise a nucleic acid sequence having at least 90% sequence identity to IGF-IR (SEQ ID NO: 63). Optionally, the plasmid can comprise a nucleic acid sequence having at least 95% sequence identity to IGF-IR (SEQ ID NO: 63). Optionally, the plasmid can comprise a nucleic acid sequence having at least 99% sequence identity to IGF-IR (SEQ ID NO: 63). In some cases, the plasmid can comprise a nucleic acid sequence having 100% sequence identity to IGF-IR (SEQ ID NO: 63). In some cases, the plasmid can comprise a nucleic acid sequence consisting of 100% sequence identity to IGF-IR (SEQ ID NO: 63).

[0295] Optionally, the isolated and purified plasmid can contain at least one nucleic acid sequence encoding a polypeptide having at least 70% sequence identity to an epitope sequence selected from SEQ ID NOs: 54, 73, 85, and 87. The plasmid can contain at least one nucleic acid sequence encoding a polypeptide having at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to an epitope sequence selected from SEQ ID NOs: 54, 73, 85, and 87. The plasmid can contain at least one nucleic acid sequence encoding a polypeptide having at least 80% sequence identity to an epitope sequence selected from SEQ ID NOs: 54, 73, 85, and 87. The plasmid can contain at least one nucleic acid sequence encoding a polypeptide having at least 90% sequence identity to an epitope sequence selected from SEQ ID NOs: 54, 73, 85, and 87. The plasmid can contain at least one nucleic acid sequence encoding a polypeptide having at least 95% sequence identity to an epitope sequence selected from SEQ ID NOs: 54, 73, 85, and 87. The plasmid can contain at least one nucleic acid sequence encoding a polypeptide having at least 99% sequence identity to an epitope sequence selected from SEQ ID NOs: 54, 73, 85, and 87. The plasmid can contain at least one nucleic acid sequence encoding a polypeptide having 100% sequence identity to an epitope sequence selected from SEQ ID NOs: 54, 73, 85, and 87. The plasmid can contain at least one nucleic acid sequence encoding a polypeptide consisting of 100% sequence identity to an epitope sequence selected from SEQ ID NOs: 54, 73, 85, and 87.

[0296] In some cases, the isolated and purified plasmid can contain at least four nucleic acid sequences, and each of the four nucleic acid sequences encodes a polypeptide that contains at least 70% sequence identity to an epitope sequence selected from SEQ ID NO: 54, 73, 85, and 87. The plasmid can contain at least four nucleic acid sequences, and each of the four nucleic acid sequences encodes a polypeptide that contains at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to an epitope sequence selected from SEQ ID NO: 54, 73, 85, and 87. The plasmid can contain at least four nucleic acid sequences, and each of the four nucleic acid sequences encodes a polypeptide that contains at least 80% sequence identity to an epitope sequence selected from SEQ ID NO: 54, 73, 85, and 87. The plasmid can contain at least four nucleic acid sequences, and each of the four nucleic acid sequences encodes a polypeptide that contains at least 90% sequence identity to an epitope sequence selected from SEQ ID NO: 54, 73, 85, and 87. The plasmid can contain at least four nucleic acid sequences, and each of the four nucleic acid sequences encodes a polypeptide that contains at least 95% sequence identity to an epitope sequence selected from SEQ ID NO: 54, 73, 85, and 87. The plasmid can contain at least four nucleic acid sequences, and each of the four nucleic acid sequences encodes a polypeptide that contains at least 99% sequence identity to an epitope sequence selected from SEQ ID NO: 54, 73, 85, and 87. The plasmid can contain at least four nucleic acid sequences, and each of the four nucleic acid sequences encodes a polypeptide that contains 100% sequence identity to an epitope sequence selected from SEQ ID NO: 54, 73, 85, and 87. The plasmid can contain at least four nucleic acid sequences, and each of the four nucleic acid sequences encodes a polypeptide consisting of 100% sequence identity to an epitope sequence selected from SEQ ID NO: 54, 73, 85, and 87. In some cases, the at least four nucleic acid sequences independently encode polypeptides to an epitope sequence selected from SEQ ID NO: 54, 73, 85, and 87.In other cases, at least four nucleic acid sequences encode different polypeptides for epitope sequences selected from SEQ ID NOs: 54, 73, 85 and 87.

[0297] In some cases, the plasmid can contain four nucleic acid sequences, each of which encodes a polypeptide having at least 70% sequence identity to an epitope sequence selected from SEQ ID NOs: 54, 73, 85, and 87. The plasmid can contain four nucleic acid sequences, each of which encodes a polypeptide having at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to an epitope sequence selected from SEQ ID NOs: 54, 73, 85, and 87. The plasmid can contain four nucleic acid sequences, each of which encodes a polypeptide having at least 80% sequence identity to an epitope sequence selected from SEQ ID NOs: 54, 73, 85, and 87. The plasmid can contain four nucleic acid sequences, each of which encodes a polypeptide having at least 90% sequence identity to an epitope sequence selected from SEQ ID NOs: 54, 73, 85, and 87. The plasmid can contain four nucleic acid sequences, each of which encodes a polypeptide having at least 95% sequence identity to an epitope sequence selected from SEQ ID NOs: 54, 73, 85, and 87. The plasmid can contain four nucleic acid sequences, each of which encodes a polypeptide having at least 99% sequence identity to an epitope sequence selected from SEQ ID NOs: 54, 73, 85, and 87. The plasmid can contain four nucleic acid sequences, each of which encodes a polypeptide having 100% sequence identity to an epitope sequence selected from SEQ ID NOs: 54, 73, 85, and 87. The plasmid can contain four nucleic acid sequences, each of which encodes a polypeptide consisting of 100% sequence identity to an epitope sequence selected from SEQ ID NOs: 54, 73, 85, and 87. In some instances, the four nucleic acid sequences independently encode polypeptides to an epitope sequence selected from SEQ ID NOs: 54, 73, 85, and 87. In other instances, the four nucleic acid sequences encode different polypeptides to an epitope sequence selected from SEQ ID NOs: 54, 73, 85, and 87.

[0298] In some cases, the plasmid can include a nucleic acid sequence encoding a polypeptide having at least 70% sequence identity to SEQ ID NO: 89. In some examples, the plasmid can include a nucleic acid sequence encoding a polypeptide having at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to SEQ ID NO: 89. The plasmid can include a nucleic acid sequence encoding a polypeptide having at least 80% sequence identity to SEQ ID NO: 89. The plasmid can include a nucleic acid sequence encoding a polypeptide having at least 90% sequence identity to SEQ ID NO: 89. The plasmid can include a nucleic acid sequence encoding a polypeptide having at least 95% sequence identity to SEQ ID NO: 89. The plasmid can include a nucleic acid sequence encoding a polypeptide having at least 99% sequence identity to SEQ ID NO: 89. The plasmid can include a nucleic acid sequence encoding a polypeptide having 100% sequence identity to SEQ ID NO: 89. The plasmid can include a nucleic acid sequence encoding a polypeptide consisting of 100% sequence identity to SEQ ID NO: 89.

[0299] In some cases, the plasmid can comprise a nucleic acid sequence having at least 70% sequence identity to SEQ ID NO: 90. In some examples, the plasmid can comprise a nucleic acid sequence having at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to SEQ ID NO: 90. The plasmid can comprise a nucleic acid sequence having at least 80% sequence identity to SEQ ID NO: 90. The plasmid can comprise a nucleic acid sequence having at least 90% sequence identity to SEQ ID NO: 90. The plasmid can comprise a nucleic acid sequence having at least 95% sequence identity to SEQ ID NO: 90. The plasmid can comprise a nucleic acid sequence having at least 99% sequence identity to SEQ ID NO: 90. The plasmid can comprise a nucleic acid sequence having 100% sequence identity to SEQ ID NO: 90. The plasmid can comprise a nucleic acid sequence consisting of 100% sequence identity to SEQ ID NO: 90.

[0300] In some cases, the plasmid can comprise a nucleic acid sequence having at least 70% sequence identity to SEQ ID NO: 91. In some examples, the plasmid can comprise a nucleic acid sequence having at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to SEQ ID NO: 91. The plasmid can comprise a nucleic acid sequence having at least 80% sequence identity to SEQ ID NO: 91. The plasmid can comprise a nucleic acid sequence having at least 90% sequence identity to SEQ ID NO: 91. The plasmid can comprise a nucleic acid sequence having at least 95% sequence identity to SEQ ID NO: 91. The plasmid can comprise a nucleic acid sequence having at least 99% sequence identity to SEQ ID NO: 91. The plasmid can comprise a nucleic acid sequence having 100% sequence identity to SEQ ID NO: 91. The plasmid can comprise a nucleic acid sequence consisting of 100% sequence identity to SEQ ID NO: 91.

[0301] Optionally, a plasmid containing more than one epitope sequence can include a spacer between each epitope sequence. In some examples, the sequences of the epitopes can be encoded tandemly without using a spacer. In some examples, the sequences of the epitopes can be encoded tandemly using a spacer. In some examples, the spacer can include a sequence encoding from about 1 to about 50, from about 3 to about 40, from about 5 to about 35, or from about 10 to about 30 amino acid residues. In some cases, the spacer can include a sequence encoding about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 25, or 30 amino acid residues.

[0302] In some examples, the plasmid can contain a nucleic acid sequence encoding at least one tag. In some examples, the tag can be translated into a peptide. Any nucleic acid sequence for a tag known to those of skill in the art can be used with the plasmids described herein. For example, the tag can be a histidine tag having 3 histidine residues, a histidine tag having 4 histidine residues, a histidine tag having 5 histidine residues, a histidine tag having 6 histidine residues, or the like. Expression of the tag in a subject can be determined using any suitable technique known to those of skill in the art.

[0303] In some examples, the plasmid can be sequenced using any sequencing technique known to those of skill in the art such that the results of the sequencing technique provide nucleotide-level resolution of the entire plasmid.

[0304] In some embodiments, the composition can be a multi-antigen breast cancer vaccine or a multi-antigen ovarian cancer vaccine. For example, the multi-antigen breast cancer vaccine or the multi-antigen ovarian cancer vaccine can contain multiple antigens. In some examples, the expression of one antigen can affect the expression of different antigens. In some examples, the expression of more than one antigen can affect the expression of different antigens. In some examples, the expression of one antigen can affect the expression of more than one different antigen. In some examples, the expression of one antigen may not affect the expression of different antigens. In some examples, the expression of more than one antigen may not affect the expression of different antigens. In some examples, the expression of one antigen may not affect the expression of more than one different antigen. For example, antigenic competition can limit the immunogenicity of multi-antigen vaccines. Any technique known to those skilled in the art can be used to determine whether the immune response induced after administration of the multi-antigen vaccine is of a magnitude comparable to each antigen as a single antigen vaccine. For example, ELISPOT (e.g., for the secretion of IFNγ) can determine the magnitude of the immune response. In some examples, ELISPOT can detect rodent, non-human primate, or human peptides. In some cases, the multi-antigen breast or ovarian cancer vaccine can contain multiple epitopes derived from multiple antigens selected from survivin, HIF-1α, IGF-1R, and / or IGFBP-2. Nucleic acid

[0305] An isolated nucleic acid molecule is a nucleic acid molecule that has been removed from its natural environment (i.e., subjected to human manipulation), where the natural environment is the genome or chromosome in which the nucleic acid molecule is naturally found. As such, "isolated" does not necessarily reflect the degree to which the nucleic acid molecule has been purified, but rather indicates that the molecule is not part of the entire genome or chromosome in which the nucleic acid molecule is naturally found. An isolated nucleic acid molecule can contain a gene. An isolated nucleic acid molecule containing a gene is not a fragment of the chromosome containing such gene, but rather contains the coding region and regulatory regions associated with the gene, but does not contain additional genes naturally found on the same chromosome. An isolated nucleic acid molecule can also contain an additional nucleic acid that is not normally adjacent to the specified nucleic acid sequence in its natural state (i.e., at the 5' and / or 3' ends of the sequence) (i.e., a heterologous sequence).

[0306] Isolated nucleic acid molecules can include DNA, RNA, or hybrids in which both genomic and cDNA nucleic acids can contain a combination of deoxyribonucleotides and ribonucleotides, as well as combinations of bases including uracil, adenine, thymine, cytosine, guanine, inosine, xanthine, hypoxanthine, isocytosine, and isoguanine. Nucleic acids can be obtained by chemical synthesis methods or recombinant methods. The phrase "nucleic acid molecule" primarily refers to a physical nucleic acid molecule, and the phrase "nucleic acid sequence" primarily refers to the sequence of nucleotides in a nucleic acid molecule, but the two phrases can be used interchangeably, particularly with respect to nucleic acid molecules or nucleic acid sequences that can encode a protein or a domain of a protein.

[0307] A nucleic acid molecule can refer to at least two nucleotides covalently linked together. The nucleic acids described herein can contain phosphodiester bonds, but in some examples, as outlined below (e.g., in the construction of probes such as primers and labeled probes), for example, phosphoramidites (Beaucage et al., Tetrahedron Volume 49 (No. 10): page 1925 (1993) and the references cited therein; Letsinger, J. Org. Chem. Volume 35: page 3800 (1970); Sprinzl et al., Eur. J. Biochem. Volume 81: page 579 (1977); Letsinger et al., Nucl. Acids Res. Volume 14: page 3487 (1986); Sawai et al., Chem. Lett. 805 (1984), Letsinger et al., J. Am. Chem. Soc. Volume 110: page 4470 (1988); and Pauwels et al., Chemica Scripta Volume 26: page 141 (1986)), phosphorothioate (Mag et al., Nucleic Acids Res. Volume 19: page 1437 (1991); and U.S. Patent No. 5,644,048), phosphorodithioate (Brü et al., J. Am. Chem. Soc. Volume 111: page 2321 (1989)), O-methylphosphoroamidite linkages (see Eckstein, Oligonucleotides and Analogues: A Practical Approach, Oxford University Press) and peptide nucleic acid (herein also referred to as "PNA") backbones and linkages (all of which are incorporated by reference, see Egholm, J. Am. Chem. Soc. Volume 114: page 1895 (1992); Meier et al., Chem. Int. Ed. Engl. Volume 31: page 1008 (1992); Nielsen, Nature, Volume 365: page 566 (1993); Carlsson et al., Nature Volume 380: page 207 (1996)) having alternative backbones include nucleic acid analogs that can be. Other analog nucleic acids are locked nucleic acid (herein also referred to as "LNA"), Koshkin et al., J. Am. Chem. Soc. Volume 120: page 13 Pages 252 - 253 (1998); positive backbone (Denpcy et al., Proc. Natl. Acad. Sci. USA 92: 6097 (1995)); non - ionic backbone (U.S. Pat. Nos. 5,386,023, 5,637,684, 5,602,240, 5,216,141 and 4,469,863; Kiedrowshi et al., Angew. Chem. Intl. Ed. English 30: 423 (1991); Letsinger et al., J. Am. Chem. Soc. 110: 4 470 (1988); Letsinger et al., Nucleotide & Nucleotide 13: 1597 (1994); Chapters 2 and 3, ASC Symposium Series 580 "Carbohydrate Modifications in Antisense Research", eds. Y. S. Sanghui and P. Dan Cook; Mesmaeker et al., Bioorganic & Medicinal Chem. Lett. 4: 395 (199 4); Jeffs et al., J. Biomolecular NMR 34: 17 (1994); Tetrahedron Lett. 37: 743 (1996)), as well as U.S. Pat. Nos. 5,235,033 and 5,034,506 and Chapters 6 and 7, ASC Symposium Series 580 "Carbohydrate Modifications in Antisense Research", Y. S. Sanghui and P. Dan Cook eds., including non - ribose backbones described therein, bicyclic structures It includes analog nucleic acids having [the relevant feature]. Nucleic acids containing one or more carbocyclic sugars are also included within the definition of nucleic acids (see Jenkins et al., Chem. Soc. Rev. (1995), pp. 169-176). Several types of nucleic acid analogs are described in Rawls, C & E News, June 2, 1997, p. 35. "Locked nucleic acid" is also included within the definition of nucleic acid analogs. LNA is a class of nucleic acid analogs in which the ribose ring is "locked" by a methylene bridge connecting the 2'-O atom and the 4'-C atom. All of these references are hereby expressly incorporated by reference into this specification. These modifications of the ribose-phosphate backbone can be made to increase the stability and half-life of such molecules in a physiological environment. For example, PNA:DNA and LNA-DNA hybrids can exhibit higher stability and thus can be used in some embodiments. The target nucleic acid can be single-stranded or double-stranded as specified, or can contain portions of both double-stranded or single-stranded sequences. Depending on the application, the nucleic acid can be DNA (including, for example, genomic DNA, mitochondrial DNA, and cDNA), RNA (including, for example, mRNA and rRNA), or a hybrid in which the nucleic acid contains any combination of deoxyribonucleotides and ribonucleotides, and can be any combination of bases including uracil, adenine, thymine, cytosine, guanine, inosine, xathanine, hypoxathanine, isocytosine, isoguanine, etc.

[0308] A recombinant nucleic acid molecule can comprise at least one of any nucleic acid sequence encoding any one or more of the proteins described herein, operably linked to at least one of any transcriptional control sequences capable of effectively regulating the expression of the nucleic acid molecule(s) in the cell(s) to be transfected. The term "nucleic acid molecule" mainly refers to a physical nucleic acid molecule, and the term "nucleic acid sequence" mainly refers to the sequence of nucleotides in a nucleic acid molecule. However, the two terms can be used interchangeably, especially with respect to a nucleic acid molecule or nucleic acid sequence capable of encoding a protein. In addition, the term "recombinant molecule" mainly refers to a nucleic acid molecule operably linked to a transcriptional control sequence, but can be used interchangeably with the term "nucleic acid molecule" administered to an animal.

[0309] A recombinant nucleic acid molecule comprises a recombinant vector, which is an isolated nucleic acid sequence operably linked to an isolated nucleic acid molecule encoding a fusion protein of the present invention, which enables the recombinant production of the fusion protein and can deliver the nucleic acid molecule into a host cell according to the present invention, typically a heterologous sequence. Such a vector can contain nucleic acid sequences that are not naturally found adjacent to the isolated nucleic acid molecule to be inserted into the vector. The vector can be either RNA or DNA, and can be either prokaryotic or eukaryotic, and is preferably a virus or a plasmid in the present invention. The recombinant vector can be used in the cloning, sequencing and / or other manipulation of nucleic acid molecules, and can be used in the delivery of such molecules (e.g., in a DNA composition or a composition based on a viral vector). The recombinant vector is preferably used in the expression of nucleic acid molecules and can also be referred to as an expression vector. A preferred recombinant vector can be expressed in a transfected host cell.

[0310] In the recombinant molecule of the present invention, the nucleic acid molecule is compatible with the host cell and is operably linked to an expression vector containing regulatory sequences such as transcriptional regulatory sequences, translational regulatory sequences, origins of replication, and other regulatory sequences that control the expression of the nucleic acid molecule of the present invention. In particular, the recombinant molecule of the present invention comprises a nucleic acid molecule operably linked to one or more expression control sequences. The phrase "operably linked" refers to the linking of a nucleic acid molecule to an expression control sequence in such a manner that the molecule is expressed when transfected (i.e., transformed, transduced, or transfected) into a host cell. Pharmaceutical composition

[0311] The immunogenic composition of the present disclosure is preferably formulated as a vaccine for in vivo administration to a subject such that it confers an antibody titer that exceeds the criterion for the antibody retention rate of each antigenic component at an acceptable percentage for the subject. Antigens having a relevant antibody titer above which the subject is considered to have seroconverted against the antigen are well known, and such titers have been published by organizations such as the WHO. Preferably, more than 80% of the statistically significant samples of the subject seroconvert, more preferably more than 90%, even more preferably more than 93%, and most preferably 96 - 100% seroconvert. Adjuvant

[0312] The immunogenic composition of the present disclosure is preferably adjuvanted. An adjuvant can be used to enhance the immune response (humoral and / or cellular) induced in a patient who has received a vaccine. Optionally, the adjuvant can induce a TH1-type response. In other cases, the adjuvant can induce a TH2-type response. In contrast to a TH2-type response, which can be characterized by the production of cytokines such as IL-4, IL-5, and IL-10, a TH1-type response can be characterized by the production of cytokines such as IFN-γ.

[0313] An adjuvant can contain stimulatory molecules such as cytokines. Non-limiting examples of cytokines include CCL20, alpha-interferon (IFN-α), beta-interferon (IFN-β), gamma-interferon, platelet-derived growth factor (PDGF), TNFα, TNFβ, granulocyte macrophage colony-stimulating factor (GM-CSF), epidermal growth factor (EGF), cutaneous T cell-attracting chemokine (CTACK), thymus-expressed chemokine (TECK), mucos...

Claims

**Claim 1**: A nucleic acid comprising a nucleotide sequence encoding a fusion peptide, wherein the amino acid sequence of the fusion peptide has at least 90% sequence identity to the amino acid sequence of SEQ ID NO: 39, and the amino acid sequence of the fusion peptide comprises the amino acid sequences of SEQ ID NO: 1, 16, 20, 26, and 32. **Claim 2**: The nucleic acid according to claim 1, wherein the amino acid sequence of the fusion peptide has at least 95% sequence identity to the amino acid sequence of SEQ ID NO:

39. **Claim 3**: The nucleic acid according to claim 1, wherein the amino acid sequence of the fusion peptide has at least 99% sequence identity to the amino acid sequence of SEQ ID NO:

39. **Claim 4**: The nucleic acid according to claim 1, wherein the amino acid sequence of the fusion peptide is the amino acid sequence of SEQ ID NO:

39. **Claim 5**: The nucleic acid according to any one of claims 1 to 4, wherein the nucleotide sequence is located in the pUMVC3 plasmid backbone. **Claim 6**: The nucleic acid according to any one of claims 1 to 5, wherein the nucleic acid is provided in a composition, and the composition further comprises a pharmaceutical carrier, an adjuvant, or a combination thereof. **Claim 7**: The nucleic acid according to claim 6, wherein the adjuvant is GM-CSF. **Claim 8**: The nucleic acid according to claim 7, which is capable of inducing a Th1 immune response when administered to a subject.

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