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 March 28, 2014, which is incorporated herein by reference in its entirety.

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

[0003] Cancer treatment has traditionally been achieved by surgical reduction of tumor burden followed by chemotherapy and / or radiation therapy. This strategy can reduce tumor size and, in less advanced stages, often results in complete remission. Unfortunately, the prognosis for more advanced tumors has changed little over the past 50 years, and a significant proportion of cancer-related deaths are caused by subsequent metastasis. New preventative and therapeutic treatments are needed to combat the increasing incidence of cancer.

[0004] Worldwide, more than one million people are diagnosed with breast cancer each year, and more than 400,000 people die from the disease each year. It is estimated that one in eight women will be diagnosed with breast cancer at some point in their lifetime. Preventing breast cancer from developing could have significant health and economic benefits for every individual. Billions of dollars could be saved by avoiding the need for expensive cancer-related surveillance and treatment interventions. New approaches to breast cancer prevention and treatment are needed. Summary of the Invention [Means for solving the problem]

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

[0006] In another aspect, the disclosure includes 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 HIF-1α peptide, and the first nucleotide sequence is located on the plasmid. In yet another aspect, the disclosure includes 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 a HIF-1α peptide, and the first nucleotide sequence and the second nucleotide sequence are located on one or more plasmids.

[0007]

[0003] Compositions described herein, in some aspects, include compositions 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 wherein the first nucleotide sequence is located on the plasmid. In other aspects, the present disclosure includes compositions 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 wherein the first nucleotide sequence and the second nucleotide sequence are located on one or more plasmids.

[0008]

[0003] In some embodiments, the compositions described herein include compositions 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 IGFBP-2, HER-2, and IGF-1R, and the first nucleotide sequence is located on the plasmid. In yet some other embodiments, the present disclosure includes compositions 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 IGFBP-2, HER-2, or IGF-1R, and the first nucleotide sequence and the second nucleotide sequence are located on one or more plasmids.

[0009] The compositions described herein, in some embodiments, include compositions comprising a first epitope of a first antigen expressed by a cell associated with breast cancer and a second epitope of a second antigen expressed by a cell associated with breast cancer.

[0010] In other aspects, the disclosure includes compositions 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 yet other aspects, the disclosure includes compositions 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 aspects, the present disclosure includes compositions 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 yet other aspects, the present disclosure includes compositions 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 instances, the 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 comprising 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 and second nucleotide sequences can be located on one or more isolated and purified plasmids. The first and second epitopes can be independently selected from portions of the HIF-1α peptide that contain 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 and second nucleotide sequences can be located on 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 and second nucleotide sequences can be located on one or more isolated and purified plasmids. The first and second nucleic acid sequences can be located on a first isolated and purified plasmid. The second nucleic acid sequence can be located on a second isolated and purified plasmid.

[0014] In some examples, the disclosure includes a composition comprising a first epitope of a first antigen expressed by a cell associated with breast cancer or ovarian cancer and a second epitope of a second antigen expressed by a cell 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 instances, the disclosure includes a composition comprising 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.

[0016] In some examples, the disclosure includes a composition comprising a plasmid comprising four nucleotide sequences and an excipient, 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 NOs: 54, 73, 85, and 87.

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

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

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

[0020] Optionally, described herein is 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. Described herein is a method for preventing breast cancer in a subject, the method comprising administering to the subject a composition described herein.

[0021] Optionally, described herein is 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. Described herein is a method for treating breast cancer in a subject, the method comprising administering to the subject a composition described herein.

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

[0023] Described herein, in some cases, is a method of generating an immune response in a subject with breast cancer or ovarian cancer, the method comprising administering to the subject a composition described herein.

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

[0025] The disclosure also includes 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: 1, 6, 8-10, 14-16, 20, 25-28, and 32-34. 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 comprising at least 90% sequence identity selected from SEQ ID NOs: 1, 6, 8-10, 14-16, 20, 25-28, and 32-34. 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 comprising at least 90% sequence identity to a sequence selected from SEQ ID NOs: 1, 6, 8-10, 14-16, 20, 25-28, and 32-34, and each of the nucleotides is not identical within the set of two or more nucleotide sequences.

[0026] The disclosure can include isolated and purified plasmids comprising at least one nucleotide sequence encoding a polypeptide comprising 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 plasmids can comprise a set of two or more nucleotide sequences, each of the two or more nucleotide sequences independently encoding a polypeptide comprising at least 90% sequence identity selected from SEQ ID NOs: 46-56, 60-62, or 66-75. The isolated and purified plasmids can comprise a set of two or more nucleotide sequences, each of the two or more nucleotide sequences encoding a polypeptide comprising at least 90% sequence identity 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.

[0027] Optionally, the disclosure can further include 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 NOs: 54, 73, 85 and 87. Incorporation by Reference

[0028] All publications, patents, and patent applications mentioned in this specification are herein 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 set forth with particularity 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 disclosure are utilized, and the accompanying drawings. [Brief explanation of the drawings]

[0030] [Figure 1]FIG. 1 demonstrates that Th1 and Th2 epitopes vary in functional avidity.

[0031] [Figure 2] FIG. 2 shows that Th2 suppresses the anti-tumor efficacy of Th1.

[0032] [Figure 3] FIG. 3 depicts antigen-specific IgG immunity.

[0033] [Figure 4] FIG. 4 shows population-based epitope screening.

[0034] [Figure 5] Figure 5 demonstrates the characteristics of breast cancer subjects.

[0035] [Figure 6] FIG. 6 depicts antigen-specific IFNγ responses to stem cell / EMT proteins.

[0036] [Figure 7] FIG. 7 depicts antigen-specific IL-10 responses to stem cell / EMT proteins.

[0037] [Figure 8] FIG. 8 shows that the peptide in the extended sequence verifies as a native epitope with the CD105 extended epitope (52 aa) QNGTWPREVLLVLSVNS SVFLHL QALGI PLHLAYNSSLVTFQEPPGVNTTEL (SEQ ID NO: 1).

[0038] [Figure 9] FIG. 9 shows the extended epitope of Yb-1 based on the IFNγ / IL-10 activity ratio.

[0039] [Figure 10]Figure 10 shows the magnitude and incidence of IFNγ dominance. IFNγ / IL-10 activity ratio of CDH3 antigen.

[0040] [Figure 11] Figure 11 shows the magnitude and incidence of IFNγ dominance. IFNγ / IL-10 activity ratio of HIF1α antigen.

[0041] [Figure 12] FIG. 12 depicts a chart of the simultaneous in vivo evaluation in mice.

[0042] [Figure 13] FIG. 13 shows the immunogenicity and efficacy of an exemplary Yb-1 plasmid-based vaccine in mice.

[0043] [Figure 14] Figure 14 depicts the map, immunogenicity and exemplary sequences of the compositions described herein. SEQ ID NO: 39 is illustrated in Figure 14.

[0044] [Figure 15A] FIG. 15A shows an exemplary validation of peptide-specific T cells as native epitopes.

[0045] [Figure 15B] FIG. 15B depicts a timeline of a clinical trial using the compositions described herein.

[0046] [Figure 15C] FIG. 15C depicts a study schema for Phase I of a clinical trial using the compositions described herein.

[0047] [Figure 16] FIG. 16 shows Western blot analysis of HIF1α expression in the single plasmid pHIF1α and the BCMA5 plasmid encoding five antigens.

[0048] [Figure 17] Figure 17 shows the magnitude and incidence of IFNγ dominance. IFNγ / IL-10 activity ratio of the CD105 antigen.

[0049] [Figure 18] Figure 18 shows the magnitude and incidence of IFNγ dominance. IFNγ / IL-10 activity ratio of MDM-2 antigen.

[0050] [Figure 19] Figure 19 shows the magnitude and incidence of IFNγ dominance. IFNγ / IL-10 activity ratio of SOX-2 antigen.

[0051] [Figure 20] Figure 20 shows the magnitude and incidence of IFNγ dominance. IFNγ / IL-10 activity ratio of Yb-1 antigen.

[0052] [Figure 21] FIG. 21 depicts HIF1α peptide and plasmid vaccine immunogenicity and efficacy in mice.

[0053] [Figure 22] FIG. 22 depicts CD105 peptide and plasmid vaccine immunogenicity and efficacy in mice.

[0054] [Figure 23] FIG. 23 depicts CDH3 peptide and plasmid vaccine immunogenicity and efficacy in mice.

[0055] [Figure 24] FIG. 24 depicts SOX2 peptide and plasmid vaccine immunogenicity and efficacy in mice.

[0056] [Figure 25] FIG. 25 depicts MDM2 peptide and plasmid vaccine immunogenicity and efficacy in mice.

[0057] [Figure 26] Figure 26 shows the mass of the mice 3 months after the last vaccination.

[0058] [Figure 27] Figure 27 shows the mass of the mice 10 days after the last vaccination.

[0059] [Figure 28] FIG. 28 demonstrates that the IGFBP-2 C-terminus is enriched for epitopes that induce IL-10-secreting T cells compared to the N-terminus.

[0060] [Figure 29] FIG. 29 shows that N-terminal, but not C-terminal, IGFBP-2 vaccines stimulate type I immunity and inhibit tumor growth.

[0061] [Figure 30] FIG. 30 shows that IGFBP-2 vaccine-induced Th2 suppresses the anti-tumor effect of IGFBP-2-specific Th1.

[0062] [Figure 31] FIG. 31 demonstrates that a vaccine based on a HER2 Th1 epitope can increase survival associated with the epitope being spread in patients with advanced HER2+ breast cancer.

[0063] [Figure 32] FIG. 32 demonstrates that expanded Th plasmid-based vaccines are more effective than peptide-based vaccines in generating tumor antigen-specific Th1 immunity.

[0064] [Figure 33] Figure 33 shows that persistent HER2 ICD-specific immunity was abolished >1 year after plasmid DNA-based vaccination.

[0065] [Figure 34]FIG. 34 shows IGF-1R epitopes screened for IFNγ and IL-10 T cell secretion by ELISPOT.

[0066] [Figure 35] FIG. 35 demonstrates that a multi-epitope IGF-1R vaccine inhibits the growth of implanted breast cancer.

[0067] [Figure 36] FIG. 36 shows that a multi-antigen polyepitope vaccine prevents breast cancer development in mice.

[0068] [Figure 37] FIG. 37 shows exemplary cytokine secretion patterns induced by HER2 vaccination.

[0069] [Figure 38] FIG. 38 depicts the ROC analysis of stem cell / EMT antigens.

[0070] [Figure 39] FIG. 39 depicts candidate proteins overexpressed in stem cells and / or EMT.

[0071] [Figure 40] FIG. 40 illustrates a cartoon representation of the constructs described herein.

[0072] [Figure 41] FIG. 41 shows Western blot images of IGFBP-2, survivin, HIF-1A and IGF-IR expression.

[0073] [Figure 42-1]Figures 42A-D show the antitumor effect of the multi-antigen vaccine in the ID8 ovarian cancer implant model. Mice were imaged in an IVIS Bioluminescent Imager 3 weeks after ID8-Luc implantation. Total luminous flux (photons / second) was measured at the primary implant (Figure 42A) and metastatic sites (Figure 42B). Representative animals are shown with adjuvant alone (Figure 42C) and tri-antigen vaccination (Figure 42D). [Figure 42-2] Figures 42A-D show the antitumor effect of the multi-antigen vaccine in the ID8 ovarian cancer implant model. Mice were imaged in an IVIS Bioluminescent Imager 3 weeks after ID8-Luc implantation. Total luminous flux (photons / second) was measured at the primary implant (Figure 42A) and metastatic sites (Figure 42B). Representative animals are shown with adjuvant alone (Figure 42C) and tri-antigen vaccination (Figure 42D).

[0074] [Figure 43A] Figures 43A and 43B illustrate TH1 responses as a function of protein sequence. Selective TH1-inducing sequences were identified in the N-terminus of HIF1a and the C-terminus of survivin. The average cSPW x incidence per peptide is shown by donor type. IFN-g cSPW x incidence for volunteer donors (n=20) (white) and cancer donors (n=20) (gray) is shown on the positive y-axis. IL-10 cSPW x incidence for volunteer donors (solid) and cancer donors (dotted black) is shown on the negative y-axis. Figure 43A shows TH1 responses to HIF-1A peptides. Figure 43B shows TH1 responses to survivin peptides. Vertical lines indicate selected sequences. [Figure 43B]Figures 43A and 43B illustrate TH1 responses as a function of protein sequence. Selective TH1-inducing sequences were identified in the N-terminus of HIF1a and the C-terminus of survivin. The average cSPW x incidence per peptide is shown by donor type. IFN-g cSPW x incidence for volunteer donors (n=20) (white) and cancer donors (n=20) (gray) is shown on the positive y-axis. IL-10 cSPW x incidence for volunteer donors (solid) and cancer donors (dotted black) is shown on the negative y-axis. Figure 43A shows TH1 responses to HIF-1A peptides. Figure 43B shows TH1 responses to survivin peptides. Vertical lines indicate selected sequences.

[0075] [Figure 44] Figures 44A-D show a comparison of IgG antibody expression levels in ovarian cancer patients and volunteers. IgG antibodies specific to candidate antigens are significantly elevated in ovarian cancer patients compared to volunteer controls. IgG (y-axis) in ug / ml and experimental population (x-axis) are shown for IGF-IR (Figure 44A), IGFBP-2 (Figure 44B), HIF-1A (Figure 44C), and survivin (Figure 44D). Mean and 2 standard deviations of volunteer controls (dotted line), *p<0.05; **p<0.01; ***p<0.001. DETAILED DESCRIPTION OF THE INVENTION

[0076] The present disclosure provides breast cancer vaccine and ovarian cancer vaccine compositions, often for the prevention or treatment of breast cancer or ovarian cancer. The present disclosure also provides methods for administering breast cancer vaccine or 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 may include a nucleic acid sequence encoding an epitope of a breast cancer or ovarian cancer antigen capable of eliciting an immunogenic response in a subject, a plasmid containing the sequence described herein, an adjuvant, a pharmaceutical carrier, and an inert chemical suitable for use with the pharmaceutical composition. The breast cancer or ovarian cancer antigen may be at least one of any antigens expressed in a subject who may have or develop breast cancer or ovarian cancer. In many cases, breast cancer or ovarian cancer antigens are expressed by tissues such as breast cancer cells, ovarian cancer cells, and / or breast cancer or ovarian cancer stem cells (CSCs). CSCs may exhibit the potential for self-renewal, uncontrolled growth, and drug resistance. In some examples, CSCs may express proteins (e.g., antigens), and for example, the level of protein (e.g., antigen) expression by CSCs may 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 may 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 cases, one epitope of a breast cancer or ovarian cancer antigen can be used in the composition. In other cases, more than one epitope of a breast cancer or ovarian cancer antigen can be used in the composition. In other cases, more than two antigens, more than three, more than four, more than five, more than six, more than seven, more than eight, more than nine, more than ten, more than fifteen, more than twenty, more than twenty-five, or more than thirty breast cancer or ovarian cancer antigens can be used in the composition. In some cases, the antigens can be the same. In other cases, 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 (e.g., CSCs, such as breast CSCs or ovarian CSCs) with aberrant (e.g., upregulated) expression of a protein to prevent breast cancer or ovarian cancer.

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

[0080] The compositions and methods described herein can induce an immune response in a subject. The immune response can be an immune response to 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 anything in the subject that contains the vaccine's antigen. The compositions and methods described herein can induce a type 1 (Th1) immune response in a subject. A 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 cases, the inflammatory cytokines activate another subtype of immune cell (e.g., cytotoxic T cells) that can destroy anything in the subject that contains the antigen.

[0081] The screening method described herein for identifying epitopes and binding peptides from tumor antigens can be used to screen epitopes of multiple antigens for inducing a Th1 immune response. For example, the screening method can identify epitopes from at least one tumor antigen that induce a Th1 response (e.g., preferentially induce the secretion of Th1 cytokines) against breast cancer or ovarian cancer antigens, including CSC (e.g., breast CSC or ovarian CSC) antigens described herein.

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

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

[0084] In some cases, one adjuvant can be used in the composition.In other cases, 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 cases, one pharmaceutical carrier can be used in the composition.In other cases, 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 cases, one chemical substance can be used in the composition. In other examples, more than 1 chemical, more than 2 chemicals, more than 3 chemicals, more than 4 chemicals, more than 5 chemicals, more than 6 chemicals, more than 7 chemicals, more than 8 chemicals, more than 9 chemicals, or more than 10 chemicals can be used in the composition.

[0085] The present disclosure further describes methods for administering a breast cancer or ovarian cancer vaccine to a subject. In some examples, the method can include constructing a plasmid-based vaccine targeting these antigens and determining whether administering the vaccine is safe, immunogenic, and effective in preventing breast cancer development. For example, the composition can be a multi-antigen Th1 polyepitope plasmid-based vaccine. 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 CSCs (e.g., breast CSCs or ovarian CSCs) and / or associated with epithelial to mesenchymal cell (EMT) cell transition. 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 can include nucleic acids encoding more than one antigen or more than one epitope of an antigen. In some examples, the methods can be used to determine whether the compositions described herein prevent the development of breast cancer or ovarian cancer in cancer (e.g., breast cancer or ovarian cancer) models using multiple organisms, for example, genetically similar rodents (e.g., mice), using genetically diverse rodents (e.g., mice), and in subjects who may or may not have 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). Antigen identification

[0086] The compositions and methods described herein include the identification and genetic manipulation of breast cancer or ovarian cancer antigens in pharmaceutical compositions (e.g., vaccines). While any technique known to those skilled in the art may be used to identify antigens expressed by subjects with breast cancer or ovarian cancer, in illustrative examples, suitable antigens can be identified using the methods described herein. In some examples, the method can include screening serum from the subject. In some examples, the screening can be an antibody screening. For example, the antibody screened can be an IgG antibody. In some examples, the serum can be derived from a subject with breast cancer or ovarian cancer. In other examples, the serum can be derived from a subject without breast cancer or ovarian cancer.

[0087] For example, a cancer antigen, such as a breast cancer antigen or an ovarian cancer antigen, can be a portion of a protein, a portion of a peptide, or a portion of a polyamino acid. In some examples, the portion can be a percentage of the protein, a percentage of the peptide, or a percentage of the 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 midpoint. In some examples, the moiety can be located at the N-terminus of the protein, peptide, or polyamino acid. In other examples, the moiety 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 midpoint. In some examples, the moiety can be located near the middle of the protein, peptide, or polyamino acid. In other examples, the moiety can be located near the middle of the protein, peptide, or polyamino acid. For example, the vicinity of the middle can be within 1%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50% of the length of the total protein, peptide or polyamino acid from the end.

[0088] At least one antigen can be identified and screened for suitability as an antigen in a composition (e.g., a vaccine) described herein. In some cases, one antigen can be identified and screened. In other examples, more than 1 antigen may be identified and screened for suitability in a vaccine, more than 2 antigens may be identified and screened, more than 3 antigens may be identified and screened, more than 4 antigens may be identified and screened, more than 5 antigens may be identified and screened, more than 6 antigens may be identified and screened, more than 7 antigens may be identified and screened, more than 8 antigens may be identified and screened, more than 9 antigens may be identified and screened, more than 10 antigens may be identified and screened, more than 11 antigens may be identified and screened, more than 12 antigens may be identified and screened, more than 13 antigens may be identified and screened, more than 14 antigens may be identified and screened, more than 15 antigens may be identified and screened, more than 20 antigens may be identified and screened, more than 25 antigens may be identified and screened, more than 30 antigens may be identified and screened, more than 35 antigens may be identified and screened, more than 40 antigens may be identified and screened, more than 45 antigens may be identified and screened, or more than 50 antigens may be identified and screened. In an illustrative example, five antigens can be identified and screened for suitability in a vaccine.

[0089] The antigen that is screened for suitability in vaccines can be derived from any protein that is detected in the serum of subjects with breast cancer or ovarian cancer, using screening techniques known to those skilled in the art.In some examples, screening can often be antibody screening.Protein can be any protein that is detected in the serum of subjects with breast cancer or ovarian cancer, but in exemplary examples, the protein that can be derived from antigen can be classified as stem cell protein and / or EMT protein.For example, breast cancer stem cell / EMT protein can include SOX2, YB1, CD105, MDM2, CDH3+ / - and HIF1α.In many cases, antigen can be immunogenic in both breast cancer subjects and non-breast cancer subjects. Antigen epitope mapping

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

[0091] In some examples, the epitope can be a human leukocyte antigen (HLA) class I epitope derived from 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 breast cancer or ovarian cancer antigen for cancer vaccine (e.g., breast cancer or ovarian cancer) development. 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, epitopes can be mapped using the methods described herein, in addition to the following steps: (1) determining whether the epitope binds (e.g., with high affinity) to MHC by at least one HLA allele (e.g., HLA-DR, i.e., a universal epitope); (2) determining whether the epitope stimulates IFNγ but not IL-10 secretion (e.g., from antigen-specific T cells); and (3) determining whether T cells can recognize peptides (e.g., epitopes) processed by antigen-presenting cells (APCs), i.e., whether the epitope is a native 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 cells can be exogenous T cells 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 cases, the protein can be endogenously processed. In other cases, the protein can be exogenously processed. In some cases, the protein can be endogenously processed by autologous APC. In other cases, the protein can be exogenously processed by autologous APC.

[0092] In all examples, the peptide is an epitope mapped from the antigen and can be identified using the methods described herein for peptide epitope selection. In some examples, the epitope can be derived from a human protein, which 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 to at least one epitope to be synchronized, 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 synchronize 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 animal) in need thereof.

[0093] In some examples, the peptide is positioned within a portion of a protein, peptide, or polyamino acid such that the protein, peptide, or polyamino acid stimulates the secretion of IFNγ. In some examples, the peptide is positioned within a portion of a protein, peptide, or polyamino acid such that the protein, peptide, or polyamino acid inhibits the secretion of IFNγ. In some examples, the peptide is positioned within a portion of a protein, peptide, or polyamino acid such that the protein, peptide, or polyamino acid stimulates the secretion of IL-10. In some examples, the peptide is positioned within a portion of a protein, peptide, or polyamino acid such that the protein, peptide, or polyamino acid inhibits the secretion of IL-10. 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 instances, the amino acids that make up the peptide can be tailored to achieve the desired effect of the peptide on IFNγ secretion and / or the desired effect of the peptide on IL-10 secretion. For example, a peptide that stimulates the secretion of both IFNγ and IL-10 can be tailored such that the length of the peptide is shortened to eliminate amino acids that stimulate IL-10 secretion, so that the peptide stimulates only the secretion of IFNγ.

[0095] In some examples, the identified epitopes can be included in vaccine compositions of extended epitope vaccines. In some examples, the extended epitopes can be 40-80mer peptides. In illustrative examples, either nucleic acid or peptide sequences are juxtaposed to construct extended epitope sequences. Juxtaposition of selected peptides within a parent protein (e.g., within 10 amino acids of each other) can allow for the construction of in-tandem extended epitopes that may contain tolerizing and / or inhibitory epitopes. For example, tandem extended epitopes can contain short intervening sequences of <10 amino acids. Any of these peptides and / or extended epitopes (embodied either as the peptides themselves or as corresponding nucleic acid constructs), alone or in any combination, can be optimized into protein or plasmid-based vaccinations that specifically induce, amplify, or synchronize protective immune responses, or suppress, attenuate, or eliminate pathological responses, in subjects (human or animal) in need thereof.

[0096] In some examples, the epitope may be amino acid long. In some examples, the epitope may 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, It may be 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 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 comprising 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 comprising 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 comprising 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 includes one or more epitopes that have 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 includes one or more epitopes that have 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 includes 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 cases, the multiple epitopes are multiple consecutive epitopes. In some cases, the consecutive epitopes further comprise a linker between one or more of the epitope sequences. In some cases, the amino acid sequences of the first and second epitopes are separated by a linker amino acid sequence. In some cases, 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, wherein the additional polypeptide comprises multiple epitopes, including one or more epitopes comprising 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, wherein the additional polypeptide comprises multiple 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 the subject. In some examples, the composition is effective in eliminating a large number of cells associated with breast cancer or ovarian cancer in the subject. In some examples, the composition can be used to prevent the growth of cells associated with breast cancer or ovarian cancer in the subject.

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

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

[0104] The present disclosure also provides kits for preparing the compositions described herein, the kits including instructions for preparing the compositions. The present disclosure also provides kits for administering the compositions described herein, the kits including instructions for administering the compositions. Compositions containing epitopes for breast cancer vaccines

[0105] The compositions described herein include compositions comprising 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 nucleotide sequence encoding a second epitope of a second antigen expressed by a cell 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 compositions include 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 can include 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 is located on the plasmid. In other examples, the composition can include 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 are located on 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 can include a nucleotide sequence having at least 90% sequence identity to the nucleotide sequence of CAGAACGGCACCTGGCCCCGCGAGGTGCTGCTGGTGCTGTCCGTGAACTCCTCCGTGTTCCTGCACCTACAGGCCCTGGGCATCCCCCTGCACCTGGCCTACAACTCCTCCCTGGTGACCTTCCAGGAGCCCCCCGGCGTGAACACCACCGAGCTG (SEQ ID NO: 2); A nucleotide sequence having at least 90% sequence identity to the nucleotide sequence of: TCCGAGTCCTCCTCCACCAACCACTCCATCGGCTCCACCCAGTCCACCCCCTGCTCCACCTCCTCCATGGCC (SEQ ID NO: 3); A nucleotide sequence having at least 90% sequence identity to the nucleotide sequence of: ACCGTGTCCATGCGCCTGAACATCGTGTCCCCCGACCTGTCCGGCAAGGGCCTGGTGCTGCCCTCCGTGCTGGGCATCACCTTCGGCGCCTTCCTGATCGGCGCCCTGCTGACCGCCGCCCTGTGGTACATCTACTCCCACACCCGCGGCCCCTCCAAGCGCGAGCCCGTGGTGGCCGTGGCCGCCCCCGCCTCCTCCGAGTCCTCCTCCACCAACCACTCCATCGGCTCCACCCAGTCCACCCCCTGCTCCACCTCCTCCATGGCC (SEQ ID NO: 4). a nucleotide sequence having at least 90% sequence identity to the nucleotide sequence of: ACCGTGTCCATGCGCCTGAACATCGTGTCCCCCGACCTGTCCGGCAAGGGCCTGGTGCTGCCCTCCGTGCTGGGCATCACCTTCGGCGCCTTCCTGATCGGCGCCCTGCTGACCGCCGCCCTGTGGTACATCTACTCCCACACCCGCGCCCCCTCCAAGCGCGAGCCCGTGGTGGCCGTGGCCGCCCCCGCCTCCTCCGAGTCCTCCTCCACCAACCACTCCATCGGCTCCACCCAGTCCACCCCCTGCTCCACCTCCTCCATGGCC (SEQ ID NO: 5); a nucleotide sequence encoding an amino acid sequence having at least 90% sequence identity to the amino acid sequence of: EARMLNASIVASFVELPL (SEQ ID NO: 6); a nucleotide sequence encoding an amino acid sequence having at least 90% sequence identity to the amino acid sequence of: QNGTWPREVLLVLSVNSSVFLHLQALGIPLHLAYNSSLVTFQEPPGVNTTEL (SEQ ID NO: 7). a nucleotide sequence encoding an amino acid sequence having at least 90% sequence identity to the amino acid sequence of TVFMRLNIISPDLSGCTSKGLVLPAVLGITFGAFLIGALLTAALWYIYSHTRSPSKREPVVAVAAPASSESSSTNHSIGSTQSTPCSTSSMA (SEQ ID NO: 8); a nucleotide sequence encoding an amino acid sequence having at least 90% sequence identity to the amino acid sequence of 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 of TVSMRLNIVSPDLSGKGLVLPSVLGITFGAFLIGALLTAALWYIYSHTRAPSKREPVVAVAAPASSESSSTNHSIGSTQSTPCSTSSMA (SEQ ID NO: 10).In some examples, the composition comprises a nucleotide sequence having at least 90% sequence identity to the nucleotide sequence of GGAGTGCAGTGCAGGGCTCCAAGTACGCTGCCGACCGCAACCACTACCGCCGCTACCCACGCCGTCGCGGCCCACCCCGCAACTACCAGCAGAAC (SEQ ID NO: 11); a nucleotide sequence having at least 90% sequence identity to the nucleotide sequence of GCGTGCCCGTGCAGGGCTCCAAGTACGCCGCCGACCGCAACCACTACCGCCGCTACCCCCGCCGCCGGCCCCCCCCGCAACTACCAGCAGAAC (SEQ ID NO: 12); The nucleic acid sequence may include a nucleic acid sequence encoding an epitope of peptide Yb-1 selected from the group consisting of a nucleotide sequence having at least 90% sequence identity to the nucleotide sequence of CCGCCGCGGCCCCCCCCGCAACTACCAGCAGAAC (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). In some examples, the composition has a sequence that is at least 90% identical to the nucleotide sequence of GGCCTCAATGCGCACGGCGCAGCGCAGATGCAGCCCATGCACCGCTACGACGTGAGCGCCCTGCAGTACAACTCCATGACCAGCTCGCAGACCTACATGAACGGCTCGCCCACCTACAGCATGTCCTACTCGCAGCAGGGCACCCCTGGCATGGCTCTTGGCTCCATGGGTTCGGTG (SEQ ID NO: 17). The nucleotide sequence having identity: GGCCTGAACGCCCACGGCGCCGCCCAGATGCAGCCCATGCACCGCTACGACGTGTCCGCCCTGCAGTACAACTCCATGACCTCCTCCCAGACCTACATGAACGGCTCCCCCACCTACTCCATGTCCTACTCCCAGCAGGGCACCCCCGGCATGGCCCTGGGCTCCATGGGCTCCGTG (SEQ ID NO: 18) A nucleotide sequence having at least 90% sequence identity to the sequence: GGCCTGAACGCCCACGGCGCCGCCCAGATGCAGCCCATGCACCGCTACGACGTGTCCGCCCTGCAGTACAACTCCATGACCTCCTCCCAGACCTACATGAACGGCTCCCCCACCTACTCCATGTCCTACTCCCAGCAGGGCACCCCCGGCATGGCCCTGGGCTCCATGGGCTCCGTG (SEQ ID NO: 19); or a nucleotide sequence encoding an amino acid sequence having at least 90% sequence identity to the amino acid sequence of GLNAHGAAQMQPMHRYDVSALQYNSMTSSQTYMNGSPTYSMSYSQQGTPGMALGSMGSV (SEQ ID NO: 20). In some examples, the composition can include a nucleic acid sequence encoding an epitope of peptide SOX-2 selected from the group consisting of: AGGTCACTGAAGGAAAGGAATCCATTGAAAATCTTCCCATCCAAACGTATCTTACGAAGACACAAGAGAGATTGGGTGGTTGCTCCAATATCTGTCCCTGAAAATGGCAAGGGTCCCTTCCCACAGAGACTGAATCAGCTCAAGTCTAATAAAGATAGAGACACCAAGATTTTCTACAGCATCACGGGGCCGGGTGCAGACAGCCCACCTGAGGGTGTCTTCGCTGTAGAGAAGGACA (SEQ ID NO: 21). a nucleotide sequence having at least 90% sequence identity to the nucleotide sequence of TTGAAAATCTTCCCATCCAAACGTATCTTACGAAGACACAAGAGAGATTGGGTGGTTGCTCCAATATCTGTCCCTGAAAATGGCAAGGGTCCCTTCCCACAGAGACTGAATCAGCTCAAGTCTAATAAAGATAGAGACACCAAGATTTTCTACAGCATCACGGGGCCGGGTGCAGACAGCCCACCTGAGGGTGTCTTCGCTGTAGAGAAGGACA (SEQ ID NO: 22);A nucleotide sequence having at least 90% sequence identity to the nucleotide sequence of GCCATGCACTCCCCCCCCACCCGCATCCTGCGCCGCCGCAAGCGCGAGTGGGTGATGCCCCCCATCTTCGTGCCCGAGAACGGCAAGGGCCCCTTCCCCCAGCGCCTGAACCAGCTGAAGTCCAACAAGGACCGCGGCACCAAGATCTTCTACTCCATCACCGGCCCCGGCGCCGACTCCCCCCCCGAGGGCGTGTTCACCATCGAGAAGGAGTCC (SEQ ID NO: 23); GTGATGAACTCCCCCCCCTCCCGCATCCTGCGCCGCCGCAAGCGCGAGTGGGTGATGCCCCCCATCTCCGTGCCCGAGAACGGCAAGGGCCCCTTCCCCCAGCGCCTGAACCAGCTGAAGTCCAACAAGGACCGCGGCACCAAGCTGTTCTACTCCATCACCGGCCCCGGCGCCGACTCCCCCCCCGAGGGCGTGT a nucleotide sequence having at least 90% sequence identity to the nucleotide sequence of TCACCATCGAGAAGGAGACC (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 nucleotide sequence 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 can include 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 90% sequence identity to the amino acid sequence of ACCTACACCATGAAGGAGGTGCTGTTCTACCTGGGCCAGTACATCATGACCAAGCGCCTGTACGACGAGAAGCAGCAGCACATCGTGTACTGCTCCAACGACCTGCTGGGCGACCTGTTCGGCGTGCCCTCCTTCTCCGTGAAGGAGCACCGCAAaATCTACACCATGATCTACCGCAACCTGGTGGTGGTGAACCAGCAGGAGTCCTCCGACTCCGGCACCTCCGTGTCC (SEQ ID NO: 29); 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 TGTTCGGCGTGCCCTCCTTCTCCGTGAAGGAGCACCGCAAGATCTACGCCATGATCTACCGCAACCTGGTGGTGGTGTCCCAGCAGGACTCCGGCACCTCCCCCTCC (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 nucleotide sequence encoding an amino acid sequence having at least 90% sequence identity to the amino acid sequence of IYTMKEIIFYIGQYIMTKRLYDEKQQHIVYCSNDLLGDVFGVPSFSVKEHRKIYAMIYRNLVVVSQQDSGTSPS (SEQ ID NO: 34);

[0108] In an illustrative example, the composition comprises:ATGGCGGTACCCATGCAACTGTCCTGCTCTAGACAGAACGGCACCTGGCCCCGCGAGGTGCTGCTGGTGCTGTCCGTGAACTCCTCCGTGTTCCTGCACCTACAGGCCCTGGGCATCCCCCTGCACCTGGCCTACAACTCCTCCCTGGTGACCTTCCAGGAGCCCCCCGGCGTGAACACCACCGAGCTGAGATCCACCGGTGGAGTGCCAGTGCAGGGCTCCAAGTACGCTGCCGACCGCAACCACTACCGCCGCTACCCACGCCGTCGCGGCCCACCCCGCAACTACCAGCAGAACACGCGTGGCCTCAATGCGCACGGCGCAGCGCAGATGCAGCCCATGCACCGCTACGACGTGAGCGCCCTGCAGTACAACTCCATGACCAGCTCGCAGACCTACATGAACGGCTCGCCCACCTACAGCATGTCCTACTCGCAGCAGGGCACCCCTGGCATGGCTCTTGGCTCCATGGGTTCGGTGAGATCCCAATTGAGGTCACTGAAGGAAAGGAATCCATTGAAAATCTTCCCATCCAAACGTATCTTACGAAGACACAAGAGAGATTGGGTGGTTGCTCCAATATCTGTCCCTGAAAATGGCAAGGGTCCCTTCCCACAGAGACTGAATCAGCTCAAGTCTAATAAAGATAGAGACACCAAGATTTTCTACAGCATCACGGGGCCGGGTGCAGACAGCCCACCTGAGGGTGTCTTCGCTGTAGAGAAGGAGACAAGATCCGCCGGCGAAACCTACACCATGAAGGAGGTGCTGTTCTACCTGGGCCAGTACATCATGACCAAGCGCCTGTACGACGAGAAGCAGCAGCACATCGTGTACTGCTCCAACGACCTGCTGGGCGACCTGTTCGGCGTGCCCTCCTTCTCCGTGAAGGAGCACCGCAAAATCTACACCATGATCTACCGCAACCTGGTGGTGGTGAACCAGCAGGAGTCCTCCGACTCCGGCACCTCCGTGTCCAGATCTTAG (SEQ ID NO: 35) nucleotides,a nucleotide sequence encoding an amino acid sequence having at least 90% sequence identity to the amino acid sequence of MAVPMQLSCSRQNGTWPREVLLVLSVNSSVFLHLQALGIPLHLAYNSSLVTFQEPPGVNTTELRSTGGVPVQGSKYAADRNHYRRYPRRRGPPRNYQQNTRGLNAHGAAQMQPMHRYDVSALQYNSMTSSQTYMNGSPTYSMSYSQQGTPGMALGSMGSVRSQLRSLKERNPLKIFPSKRILRRHKRDWVVAPISVPENGKGPFPQRLNQLKSNKDRDTKIFYSITGPGADSPPEGVFAVEKETRSAGETYTMKEVLFYLGQYIMTKRLYDEKQQHIVYCSNDLLGDLFGVPSFSVKEHRKIYTMIYRNLVVVNQQESSDSGTSVSRS (SEQ ID NO: 39); a nucleotide sequence encoding an amino acid sequence having at least 90% sequence identity to the amino acid sequence of 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 MAVPMTVSMRLNIVSPDLSGK GLVLPSVLGITFGAFLIGALLTAALWYIYSHTRAPSKREPVVAVAAPASSESSSTNHSIGSTQSTPCSTSSMATGGVPVQGSKYAADRNHYRRYPRRRGPPRNYQQNTRGLNAHGAAQMQPMHRYDVSALQYNSMTSSQTYMNGSPTYSMSYSQQGTPGMALGSMGSVRSQLVMNSPPSRILRRRKREWVMPPISVPENGKGPFPQRLNQLKSNKDRGTKLFYSITGPGADSPPEGVFTIEKETRSAGEIYTMKEIIFYIGQYIMTKRLYDEKQQHIVYCSNDLLGDVFGVPSFSVKEHRKIYAMIYRNLVVVSQQDSGTSPSRS (SEQ ID NO: 42) It can include a nucleic acid sequence encoding a fusion peptide of five epitopes selected from the group consisting of nucleotide sequences encoding an amino acid sequence having at least 90% sequence identity to the amino acid sequence;

[0109] 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 including a first nucleotide sequence encoding a first epitope of a first antigen, where the first epitope is a portion of a peptide selected from IGFBP-2, HER-2, and IGF-1R, and the first nucleotide sequence is located on the plasmid. In some examples, the composition can include a first plasmid including 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, where 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, the composition has a nucleotide sequence having at least 90% sequence identity to the nucleotide sequence of ATGCTGCCGAGAGTGGGCTGCCCCGCGCTGCCGCTGCCGCCGCCGCCGCTGCTGCCGCTGCTGCCGCTGCTGCTGCTGCTACTGGGCGCGAGTGGCGGCGGCGGCGGGGCGCGCGCGGAGGTGCTGTTCCGCTGCCCGCCCTGCACACCCGAGCGCCTGGCCGCCTGCGGGCCCCCGCCGGTTGCGCCGCCCGCCGCGGTGGCCGCAGTGGCCGGAGGCGCCCGCATGCCATGCGCGGAGCTCGTCCGGGAGCCGGGCTGCGGCTGCTGCTCGGTGTGCGCCCGGCTGGAGGGCGAGGCGTGCGGCGTCTACACCCCGCGCTGCGGCCAGGGGCTGCGCTGCTATCCCCACCCGGGCTCCGAGCTGCCCCTGCAGGCGCTGGTCATGGGCGAGGGCACTTGTGAGAAGCGCCGGGACGCCGAGTATGGCGCCAGCCCGGAGCAGGTTGCAGACAATGGCGATGACCACTCAGAAGGAGGCCTGGTGGAG (SEQ ID NO: 43); ATGCTGCCCCGCCTGGGCGGCCCCGCCCTGCCCCTGCTGCTGCCCTCCCTGCTGCTGCTGCTGCTGCTGGGCGCCGGCGGCTGCGGCCCCGGCGTGCGCGCCGAGGTGCTGTTCCGCTGCCCCCCCTGCACCCCCGAGCGCCTGGCCGCCTGCGGCCCCCCCCCCGACGCCCCCTGCGCCGAGCTGGTGCGCGAGCCCGGCTGCGGCTGCTGCTCCGTGTGCGCCCGCCAGGAGGGCGAGGCCTGCGGCGTGTACATCCCCCGCTGCGCCCAGACCCTGCGCTGCTACCCCAACCCCGGCTCCGAGCTGCCCCTGAAGGCCCTGGTGACCGGCGCCGGCACCTGCGAGAAGCGCCGCGTGGGCACCACCCCCCAGCAGGTGGCCGACTCCGACGACGACCACTCCGAGGGCGGCCTGGTGGAG (SEQ ID NO: 44). A nucleotide sequence having at least 90% sequence identity to the nucleotide sequence of: ATGCTGCCCCGCCTGGGCGGCCCCGCCCTGCCCCTGCTGCTGCCCTCCCTGCTGCTGCTGCTGGGCGCCGGCGGCTGCGGCCCCGGCGTGCGCGCCGAGGTGCTGTTCCGCTGCCCCCCCTGCACCCCCGAGCGCCTGGCCGCCTGCGGCCCTGCGGCCCCCCCCCCGACGCCCCCTGCGCCGAGCTGGTGCGCGAGCCCGGCTGCGGCTGCTGCTCCGTGTGCGCCCGCCAGGAGGGCGAGGCCTGCGGCGTGTACATCCCCCGCTGCGCCCAGACCCTGCGCTGCTACCCCAACCCCGGCTCCGAGCTGCCCCTGAAGGCCCTGGTGACCGGCGCCGGCACCTGCGAGAAGCGCCGCGTGGGCACCACCCCCCAGCAGGTGGCCGACTCCGAGGACGACCACTCCGAGGGCGGCCTGGTGGAG (SEQ ID NO: 45) The sequence of the nucleotide sequence is: TMRRLLQETELVEPLTPSGAMPNQAQMRILKETELRKVKVLGSGAFGTVYKGIWIPDGENVKIPVAIKVLRENTSPKANKEILDEAYVMAGVGSPYVSRLLGICLTSTVQLVTQLMPYGCLLDHVRENRGRLGSQDLLNWCMQIAKGMSYLEDVRLVHRDLAARNVLVKSPNHVKITDFGLARLLDIDETEYHADGGKVPIKWMALESILRRRFTHQSDVWSYGVTVWELMTFGAKPYDGIPAREIPDLLEKGERLPQPPICTIDVYMIMVKCWMIDSECRPRFRELVSEFSRMARDPQRFVVIQNEDLAPGAGGMVHHRHRSSSPLPAARPAGATLERPKTLSPGKNGVVKDVFAFGGAVENPEYL (SEQ ID NO: 60) A nucleotide sequence encoding an amino acid sequence having at least 90% sequence identity to the amino acid sequence: TMRRLLQETELVEPLTPSGAVPNQAQMRILKETELRKLKVLGSGAFGTVYKGIWIPDGENVKIPVAIKVLRENTSPKANKEILDEAYVMAGVGSPYVSRLLGICLTSTVQLVTQLMPYGCLLDHVREHRGRLGSQDLLNWCVQIAKGMSYLEEVRLVHRDLAARNVLVKSPNHVKITDFGLARLLDIDETEYHADGGKVPIKWMALESILRRRFTHQSDVWSYGVTVWELMTFGAKPYDGIPAREIPDLLEKGERLPQPPICTIDVYMIMVKCWMIDSECRPRFRELVSEFSRMARDPQRFVVIQNEDLALGTGSTAHRRHRSSSPPPPIRPAGATLERPKTLSPGKNGVVKDVFAFGGAVENPEYL (SEQ ID NO: 61) A nucleotide sequence encoding an amino acid sequence having at least 90% sequence identity to the amino acid sequence; 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 a first and a second epitope independently selected from CD105, Yb-1, SOX-2, CDH3, or MDM2. In some examples, the composition can include a third epitope, wherein 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 a third and a fourth epitope, wherein 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 a third, fourth, and fifth epitope, wherein 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 a first and a second epitope independently selected from IGFBP2, HER-2, or IGF-1R. In some examples, the composition can include a third epitope, wherein the first, second, and third epitopes are independently selected from IGFBP2, HER-2, or IGF-1R.

[0112] In some cases, the composition can be administered to a subject. In some cases, the subject is in need of administration of the composition. In some cases, the composition is effective in inducing an immune response in the subject. In some cases, the composition is effective in eliminating a large number of cells associated with breast cancer in the subject. In some cases, 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 expressing atypical characteristics, pre-neoplastic 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 cases, the first and second nucleic acid sequences are purified to at least 70% purity. In some cases, the first and second nucleic acid sequences are located on a first plasmid and separated by a linker nucleic acid sequence. In some cases, the first nucleic acid sequence is adjacent to the second nucleic acid sequence on the first plasmid.

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

[0117] In some examples, the subject is selected from a human with breast cancer, a mouse with breast cancer, or a rat with 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 a 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 a 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 a ratio of IFNγ production to IL-10 production that is greater than 1. In some examples, the immune response is characterized by a ratio of IFNγ production to IL-10 production that is less than 1.

[0119] In some examples, the composition includes 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 HIF-1α peptide, and the first nucleotide sequence is located on the plasmid. In other examples, the composition includes 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 a HIF-1α peptide, and the first nucleotide sequence and the second nucleotide sequence are located on 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 differ from those listed herein. In some examples, nucleic acid sequences that are greater than 95%, 90%, 85%, 80%, 75%, 70%, 65%, 60%, 55%, or greater than 50% homologous to the nucleic acid sequences disclosed herein can be used in the compositions described herein.

[0121] The compositions described herein can, in some examples, include a composition that includes a first epitope of a first antigen expressed by a cell associated with breast cancer and a second epitope of a second antigen expressed by a cell associated with breast cancer.

[0122] In some examples, the composition can include at least a first epitope of a first antigen, where 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 include at least a first epitope of a first antigen and at least a second epitope of a second antigen, where 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 peptide CD105 has 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).In some examples, at least a first epitope of peptide Yb-1 is selected from the group consisting of 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). In some examples, at least a first epitope of peptide SOX-2 is selected from the group consisting of an amino acid sequence having at least 90% sequence identity to the amino acid sequence of GLNAHGAAQMQPMHRYDVSALQYNSMTSSQTYMNGSPTYSMSYSQQGTPGMALGSMGSV (SEQ ID NO: 20). In some examples, at least a first epitope of the peptide CDH3 has an amino acid sequence having at least 90% sequence identity to the amino acid sequence of RSLKERNPLKIFPSKRILRRHKRDWVVAPISVPENGKGPFPQRLNQLKSNKDRDTKIFYSITGPGADSPPEGVFAVEKET (SEQ ID NO: 25); or 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).In some examples, at least a first epitope of peptide MDM-2 is selected from the group consisting 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).

[0123] In some examples, the compositions described herein comprise 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 MAVPMTVSMRLNIVSPD The amino acid sequence of the fusion peptide of five epitopes selected from the group consisting of amino acid sequences having at least 90% sequence identity to the amino acid sequence of LSGKGLVLPSVLGITFGAFLIGALLTAALWYIYSHTRAPSKREPVVAVAAPASSESSSTNHSIGSTQSTPCSTSSMATGGVPVQGSKYAADRNHYRRYPRRRGPPRNYQQNTRGLNAHGAAQMQPMHRYDVSALQYNSMTSSQTYMNGSPTYSMSYSQQGTPGMALGSMGSVRSQLVMNSPPSRILRRRKREWVMPPISVPENGKGPFPQRLNQLKSNKDRGTKLFYSITGPGADSPPEGVFTIEKETRSAGEIYTMKEIIFYIGQYIMTKRLYDEKQQHIVYCSNDLLGDVFGVPSFSVKEHRKIYAMIYRNLVVVSQQDSGTSPSRS (SEQ ID NO: 42);

[0124] The compositions described herein can include compositions 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 IGFBP-2, HER-2, or IGF-1R, and the first nucleotide sequence is located on the plasmid. In some examples, the compositions 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, 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 a first epitope of the peptide IGFBP-2 is 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 an amino acid sequence having at least 90% sequence identity to the amino acid sequence of: MLPRLGGPALPLLLPSLLLLLLLGAGGCGPGVRAEVLFRCPPCTPERLAACGPPPDAPCAELVREPGCGCCSVCARQEGEACGVYIPRCAQTLRCYPNPGSELPLKALVTGAGTCEKRRVGATPQQVADSEDDHSEGGLVE (SEQ ID NO: 56). In some examples, at least a first epitope of the peptide HER-2 is directed 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 to the amino acid sequence of TMRRLLQETELVEPLTPSGAVPNQAQMRILKETELRKLKVLGSGAFGTVYKGIWIPDGENVKIPVAIKVLRENTSPKANKEILDEAYVMAGVGSPYVSRLLGICLTSTVQLVTQLMPYGCLLDHVREHRGRLGSQDLLNWCVQIAKGMSYLEEVRLVHRDLAARNVLVKSPNHVKITDFGLARLLDIDETEYHADGGKVPIKWMALESILRRRFTHQSDVWSYGVTVWELMTFGAKPYDGIPAREIPDLLEKGERLPQPPICTIDVYMIMVKCWMIDSECRPRFRELVSEFSRMARDPQRFVVIQNEDLALGTGSTAHRRHRSSSPPPPIRPAGATLERPKTLSPGKNGVVKDVFAFGGAVENPEYL (SEQ ID NO: 61). a nucleotide sequence encoding an amino acid sequence with at least 90% sequence identity; or the amino acid sequence of TMRRLLQETELVEPLTPSGAMPNQAQMRILKETELRKVKVLGSGAFGTVYKGIWIPDGENVKIPVAIKVLRENTSPKANKEILDEAYVMAGVGSPYVSRLLGICLTSTVQLVTQLMPYGCLLDHVREHRGRLGSQDLLNWCVQIAKGMSYLEDVRLVHRDLAARNVLVKSPNHVKITDFGLARLLDIDETEYHADGGKVPIKWMALESILRRRFTHQSDVWSYGVTVWELMTFGAKPYDGIPAREIPDLLEKGERLPQPPICTIDVYMIMVKCWMIDSECRPRFRELVSEFSRMARDPQRFVVIQNEDLTPGTGSTAHRRHRSSSPLPPVRPAGATLERPKTLSPGKNGVVKDVFAFGGAVENPEYL (SEQ ID NO: 62) The amino acid sequence is selected from the group consisting of nucleotide sequences encoding amino acid sequences having at least 90% sequence identity to the In some examples, the nucleic acid sequence encoding the peptide IGF-IR epitope 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 EIATLAEQPYQGLSNEQVLRFVMEGGLLDKPDNCPDMLFELMRMCWQYNPKMRPSFLEHKAENGPGPGVLVLRASFDERQPYAHMNGGRKNERALP (SEQ ID NO: 73); WSFGVVLWEIATLAEQPYQGLSNEQVLRFVMEGGLLDKPDNCPDMLFELMRMCWQYNPKMRPSFLEHKAENGPGPGVLVLRAS an amino acid sequence having at least 90% sequence identity to the amino acid sequence of: WSFGVVLWEIATLAEQPYQGLSNEQVLRFVMEGGLLDKPDNCPDMLFELMRMCWQYNPKMRPSFLEHKAENGPGVLVLRASFDERQPYAHMNGGRANERALP (SEQ ID NO: 75); or an amino acid sequence having at least 90% sequence identity to the amino acid sequence of: WSFGVVLWEIATLAEQPYQGLSNEQVLRFVMEGGLLDKPDNCPDMLFELMRMCWQYNPKMRPSFLEHKAENGPGVLVLRASFDERQPYAHMNGGRANERALP (SEQ ID NO: 75).

[0125] The compositions described herein include an amino acid sequence having at least 90% sequence identity to the amino acid sequence of (SEQ ID NO: 79);an amino acid sequence having at least 90% sequence identity to the amino acid sequence of (SEQ ID NO: 80);Or, it may further comprise 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 (SEQ ID NO: 81);

[0126] In some examples, the composition comprises a first and a second epitope independently selected from CD105, Yb-1, SOX-2, CDH3, or MDM2. In some examples, the composition further comprises a third epitope, wherein 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 a third and a fourth epitope, wherein 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 a third, fourth, and fifth epitope, wherein 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 a first and a second epitope independently selected from IGFBP2, HER-2, or IGF-1R. In some examples, the composition further comprises a third epitope, wherein the first, second, and third epitopes are independently selected from IGFBP2, HER-2, or IGF-1R.

[0128] In some examples, the composition can include at least a first epitope of a first antigen, and the first epitope is a peptide portion derived from HIF-1α. In some examples, the composition can include 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 may comprise 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 may comprise at least a first epitope of a 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 can differ from those listed herein: CD105, HIF1α, MDM2, Yb1, SOX-2, HER-2, IGFBP2, IGF-1R, and CDH3. In some instances, amino acid sequences that are greater than 95%, 90%, 85%, 80%, 75%, 70%, 65%, 60%, 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 a first plasmid. In some examples, the second nucleic acid sequence is located on a 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 a 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 a 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 a ratio of IFNγ production to IL-10 production that is greater than 1. In some examples, the immune response is characterized by a ratio of IFNγ production to IL-10 production that is less than 1.

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

[0136] In some examples, the subject is selected from the group consisting of a human with breast cancer, a mouse with breast cancer, and a rat with 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 atypical characteristics, pre-neoplastic 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 cases, at least the first epitope is contained in a pharmaceutical composition. In some cases, at least the first epitope is contained in a pharmaceutical composition further comprising a pharmaceutical carrier. In some cases, at least the first epitope is contained in a pharmaceutical composition further comprising a pharmaceutical carrier and an adjuvant. In some cases, at least the first epitope is contained in a pharmaceutical composition further comprising an adjuvant. In some cases, the composition further comprises an adjuvant and a pharmaceutical carrier. In some cases, 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, provided herein is a method for preventing breast cancer in a subject, the method comprising administering a composition described herein to the subject. In some examples, provided herein is a method for treating breast cancer in a subject, the method comprising administering a composition described herein to the subject. In some examples, the administering step further comprises delivering at least one dose of a composition described herein to the subject. In some examples, the administering step further comprises delivering a 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 with breast cancer, a mouse with breast cancer, and a rat with 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 kits for preparing the compositions described herein, the kits including instructions for preparing the compositions. The present disclosure also provides kits for administering the compositions described herein, the kits including instructions for administering the compositions. Compositions comprising epitopes 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 compositions 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 compositions 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 compositions 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 compositions 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 comprise 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 comprise a plasmid comprising at least one nucleotide sequence encoding a polypeptide consisting of 100% sequence identity over the entire 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 comprising at least 70% sequence identity to at least 20 consecutive amino acids of SEQ ID NO:85. At least one nucleotide sequence can encode a polypeptide comprising at least 80% sequence identity to at least 20 consecutive amino acids of SEQ ID NO:85. At least one nucleotide sequence can encode a polypeptide comprising at least 90% sequence identity to at least 20 consecutive amino acids of SEQ ID NO:85. At least one nucleotide sequence can encode a polypeptide comprising at least 95% sequence identity to at least 20 consecutive amino acids of SEQ ID NO:85. At least one nucleotide sequence can encode a polypeptide comprising at least 99% sequence identity to at least 20 consecutive amino acids of SEQ ID NO:85. At least one nucleotide sequence can encode a polypeptide comprising at least 100% sequence identity to at least 20 consecutive amino acids of SEQ ID NO:85.

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

[0144] In some cases, the composition includes 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 NOs: 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 NOs: 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 NOs: 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 consecutive 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 consecutive 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 consecutive 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 consecutive 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 consecutive amino acids of SEQ ID NO: 85.

[0146] Optionally, one of the four nucleotide sequences can encode a polypeptide comprising 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 comprising 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 comprising 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 comprising 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 comprising 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 in tandem within the plasmid. The four nucleotide sequences can be separated by a linker nucleic acid sequence. The linker nucleic acid sequence 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 linker amino acid sequence 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 underlined in Figure 14 (SEQ ID NO: 14).

[0148] Optionally, the composition can further comprise at least one additional isolated plasmid. Optionally, the composition can further comprise 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 compositions described herein can include a composition comprising a plasmid comprising a nucleotide sequence encoding a polypeptide comprising at least 80% sequence identity to SEQ ID NO:89 and an excipient. The composition can include a plasmid comprising a nucleotide sequence encoding a polypeptide comprising at least 90% sequence identity to SEQ ID NO:89. The composition can include a plasmid comprising a nucleotide sequence encoding a polypeptide comprising at least 95% sequence identity to SEQ ID NO:89. The composition can include a plasmid comprising a nucleotide sequence encoding a polypeptide comprising 100% sequence identity to SEQ ID NO:89. The composition can include a plasmid comprising a nucleotide sequence encoding a polypeptide consisting of 100% sequence identity to SEQ ID NO:89.

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

[0152] The composition can be formulated for treating 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 a ratio of type I cytokine production to type II cytokine production of greater than 1. The immune response can be characterized by a ratio of type I cytokine production to type II cytokine production of less than 1. The immune response can be characterized by a ratio of IFNγ production to IL-10 production of greater than 1. The immune response can be characterized by a ratio of IFNγ production to IL-10 production of less than 1.

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

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

[0156] Often, the compositions will be formulated for subcutaneous, intramuscular, or intradermal administration.

[0157] The present disclosure also provides kits for preparing the compositions described herein, the kits including instructions for preparing the compositions. The present disclosure also provides kits for administering the compositions described herein, the kits including instructions for administering the compositions. Plasmids for pharmaceutical compositions

[0158] In some examples, the epitope can be derived from a human protein that can be directly used in a peptide-based vaccine. In other examples, the epitope can be derived from a human protein, and the encoding 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 the immune response to at least one epitope to be synchronized, amplified, attenuated, suppressed, or eliminated in 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 synchronize 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 pathological responses in a subject (e.g., a human or animal) in need thereof.

[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). Any suitable plasmid known to those skilled in the art for pharmaceutical use for the expression of a nucleic acid sequence can be used. Any plasmid backbone (e.g., vector) can be used in the compositions described herein.In some cases, commercially available plasmid backbone can be used.For example, plasmid pUMVC3 can be used.In some cases, commercially available plasmid backbone can be modified, mutated, genetically engineered or cloned before use.In other cases, non-commercially available plasmid backbone can be used.

[0160] Prior to insertion of the nucleic acid sequence of at least one epitope, the plasmid backbone may be selected from the group consisting of 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 0kB, approximately 5.2kB, approximately 5.4kB, approximately 5.6kB, approximately 5.8kB, approximately 6.0kB, approximately 6.2kB, approximately 6.4kB, approximately 6.6kB, approximately 6.8kB, approximately 7.0kB, approximately 7.2kB, approximately 7.4kB, approximately 7.6kB, approximately 7.8kB, approximately 8.0kB, approximately 8.2kB, approximately 8.4kB, approximately 8.6kB, approximately 8.8kB, approximately 9.0kB, approximately 9.2kB, approximately 9.4kB, approximately 9.6kB, approximately 9.8kB, approximately 10.0kB, approximately 10. 2kB, approximately 10.4kB, approximately 10.6kB, approximately 10.8kB, approximately 11.0kB, approximately 11.2kB, approximately 11.4kB, approximately 11.6kB, approximately 11.8kB, approximately 12.0kB, approximately 12.2kB, approximately 12.4kB, 12.6kB, 12.8kB, 13.0kB, 13.2kB, 13.4kB, 13.6kB, 13.8kB, 14kB, 14.5kB, 15kB, 15.5kB, 16kB, 1 The length of the plasmid may be less than 6.5 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 an illustrative example, prior to 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, three, four, five, six, seven, eight, nine, ten, eleven, twelve, thirteen, fourteen, fifteen, sixteen, seventeen, eighteen, nineteen, twenty, 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 genome, 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, short hairpin, short interfering, messenger, transfer, ribosomal, or the like. 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, so that the epitope expressed by the nucleic acid can induce an immune response in the subject.In some examples, the subject can be a rodent, a non-human primate, or a 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 skilled in the art.The nucleic acid encoding the epitope of the plasmid can be cloned into a plasmid backbone using methods and techniques known to those skilled 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 derived from human can be used to express CD105 in human.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 derived from non-human can be used to express CD105 in human.

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

[0166] In some cases, 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 derived from human can be used to express CD105 in human.In other cases, 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 derived from non-human can be used to express CD105 in human.

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

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

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

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

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

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

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

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

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

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

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

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

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

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

[0181] In some cases, the nucleic acid sequence for expressing antigenic epitope can be 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 cases, the nucleic acid sequence encoding epitope can be 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 can be used to express SOX-2 in a subject.

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

[0183] In some cases, the nucleic acid sequence for expressing antigenic epitope can be 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 cases, the nucleic acid sequence encoding epitope can be 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 can be used to express 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 a human can be used to express HER-2 in a human. 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 a non-human can be used to express HER-2 in a human.

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

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

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

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

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

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

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

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

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

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

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

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

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

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

[0200] The compositions described herein include compositions comprising a first plasmid containing a first nucleotide sequence encoding a first epitope of a first antigen expressed by a cell associated with breast cancer, and a second nucleotide sequence encoding a second epitope of a second antigen expressed by a cell 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 compositions may include nucleic acids encoding epitopes from the following proteins: CD105, HIF1α, MDM2, Yb-1, SOX-2, HER-2, IGFBP2, IGF-1R, and CDH3.

[0201] In some examples, the composition can include 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 is located on the plasmid. In other examples, the composition can include 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 are located on one or more plasmids.

[0202] 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 a sequence identity that is at least 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100%, 101%, 102%, 103%, 104%, 105%, 106%, 107%, 108%, 109%, 110%, 111%, 112%, 113%, 114%, 115%, 116%, 117%, 118%, 119%, 120%, 121%, 122%, 123%, 124%, 125%, 126%, 127%, 128%, 129%, 130%, 131%, 132%, 133%, 134%, 135%, 136%, 137%, 138%, 139%, 140%, 141%, 142%, 143%, 144%, 145%, 146%, 147%, 148%, 149%, 150%, 151%, 152%, 153%, 154%, 155%, 156%, 157%, 158%, 159%, 160%, 161%, 162%, 163%, 164%, 165%, 166%, 167%, 168%, 169%, 170%, 171%, 172%, 173%, 174%, 175%, 176%, 177%, 178%, 179%, 180%, 181%, 182%, 183%, 184%, 185%, 186%, 187%, Nucleotide sequences with %, 98%, 99% or 100% sequence identity; ACCGTGTTCATGCGCCTGAACATCATCTCCCCCGACCTGTCCGGCTGCACCTCCAAGGGCCTGGTGCTGCCCGCCGTGCTGGGCATCACCTTCGGCGCCTTCCTGATCGGCGCCCTGCTGACCGCCGCCCTGTGGTACATCTACTCCCACACCCGCTCCCCCTCCAAGCGCGAGCCCGTGGTGGCCGTGGCCGCCCCCGCCTCCT 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 CCGAGTCCTCCTCCACCAACCACTCCATCGGCTCCACCCAGTCCACCCCCTGCTCCACCTCCTCCATGGCC (SEQ ID NO: 3); ACCGTGTCCATGCGCCTGAACATCGTGTCCCCCGACCTGTCCGGCAAGGGCCTGGTGCTGCCCT At least 50%, 60%, 70% to the nucleotide sequence of CCGTGCTGGGCATCACCTTCGGCGCCTTCCTGATCGGCGCCCTGCTGACCGCCGCCCTGTGGTACATCTACTCCCACACCCGCGGCCCCTCCAAGCGCGAGCCCGTGGTGGCCGTGGCCGCCCCCGCCTCCTCCGAGTCCTCCTCCACCAACCACTCCATCGGCTCCACCCAGTCCACCCCCTGCTCCACCTCCTCCATGGCC (SEQ ID NO: 4). 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 ACCGTGTCCATGCGCCTGAACATCGTGTCCCCCGACCTGTCCGGCAAGGGCCTGGTGCTGCCCTCCGTGCTGGGCATCACCTTCGGCGCCTTCCTGATCGGCGCCCTGCTGACCGCCGCCCTGTGGTACATCTACTCCCACACCCGCGCCCCCTCCAAGCGCGAGCCCGTGGTGGCCGTGGCCGCCCCCGCCTCCTCCGAGTCCTCCTCCACCAACCACTCCATCGGCTCCACCCAGTCCACCCCCTGCTCCACCTCCTCCATGGCC (SEQ ID NO: 5). a nucleotide sequence having 0%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity; 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 EARMLNASIVASFVELPL (SEQ ID NO: 6); 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 QNGTWPREVLLVLSVNSSVFLHLQALGIPLHLAYNSSLVTFQEPPGVNTTEL (SEQ ID NO: 1); TVFMRLNIISPDLSGCTSKGLV 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 LPAVLGITFGAFLIGALLTAALWYIYSHTRSPSKREPVVAVAAPASSESSSTNHSIGSTQSTPCSTSSMA (SEQ ID NO: 8); TVSMRLNIV 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 SPDLSGKGLVLPSVLGITFGAFLIGALLTAALWYIYSHTRGPSKREPVVAVAAPASSESSSTNHSIGSTQSTPCSTSSMA (SEQ ID NO: 9);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 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 GGAGTGCCAGGGCTCCAAGTACGCTGCCGACCGCAACCACTACCGCCGCTACCCACGCCGTCGCGGCCCACCCCGCAACTACCAGCAGAAC (SEQ ID NO: 11); or at least 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95% or 100% sequence identity to the nucleotide sequence of GCGTGCCCGTGCAGGGCTCCAAGTACGCCGCCGACCGCAACCACTACCGCCGCTACCCCCGCCGCCGGCCCCCCCCGCAACTACCAGCAGAAC (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 GCGTGCCCGTGCAGGGCTCCAAGTACGCCGCCGACCGCAACCACTACCGCCGCTACCCCCGCCGCCGCGGCCCCCCCCGCAACTACCAGCAGAAC (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 comprises at least the nucleic acid sequence of GGCCTCAATGCGCACGGCGCAGCGCAGATGCAGCCCATGCACCGCTACGACGTGAGCGCCCTGCAGTACAACTCCATGACCAGCTCGCAGACCTACATGAACGGCTCGCCCACCTACAGCATGTCCTACTCGCAGCAGGGCACCCCTGGCATGGCTCTTGGCTCCATGGGTTCGGTG (SEQ ID NO: 17); a nucleotide sequence having 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the nucleotide sequence of GGCCTGAACGCCCACGGCGCCGCCCAGATGCAGCCCATGCACCGCTACGACGTGTCCGCCCTGCAGTACAACTCCATGACCTCCTCCCAGACCTACATGAACGGCTCCCCCACCTACTCCATGTCCTACTCCCAGCAGGGCACCCCCGGCATGGCCCTGGGCTCCATGGGCTCCGTG (SEQ ID NO: 18). 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 GGCCTGAACGCCCACGGCGCCGCCCAGATGCAGCCCATGCACCGCTACGACGTGTCCGCCCTGCAGTACAACTCCATGACCTCCTCCCAGACCTACATGAACGGCTCCCCCACCTACTCCATGTCCTACTCCCAGCAGGGCACCCCCGGCATGGCCCTGGGCTCCATGGGCTCCGTG (SEQ ID NO: 18); a nucleotide sequence having 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); 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% identity to the nucleotide sequence of AGGTCACTGAAGGAAAGGAATCCATTGAAAATCTTCCCATCCAAACGTATCTTACGAAGACACAAGAGAGATTGGGTGGTTGCTCCAATATCTGTCCCTGAAAATGGCAAGGGTCCCTTCCCACAGAGACTGAATCAGCTCAAGTCTAATAAAGATAGAGACACCAAGATTTTCTACAGCATCACGGGGCCGGGTGCAGACAGCCCACCTGAGGGTGTCTTCGCTGTAGAGAAGGACA (SEQ ID NO: 21). a nucleotide sequence having sequence identity at least 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% to the nucleotide sequence of TTGAAAATCTTCCCATCCAAACGTATCTTACGAAGACACAAGAGAGATTGGGTGGTTGCTCCAATATCTGTCCCTGAAAATGGCAAGGGTCCCTTCCCACAGAGACTGAATCAGCTCAAGTCTAATAAAGATAGAGACACCAAGATTTTCTACAGCATCACGGGGCCGGGTGCAGACAGCCCACCTGAGGGTGTCTTCGCTGTAGAGAAGGACA (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); GTGATGAACTCCCCCCCCTCCCGCATCCTGCGCCGCCGCAAGCGCGAGTGGGTGATGCCCCCCATCTCCGTGCCCGAGAACGGCAAGGGCCCCTTCC A nucleic acid sequence encoding 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 CCCAGCGCCTGAACCAGCTGAAGTCCAACAAGGACCGCGGCACCAAGCTGTTCTACTCCATCACCGGCCCCGGCGCCGACTCCCCCCCCGAGGGCGTGTTCACCATCGAGAAGGAGACC (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); 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). In some examples, the composition has at least a homology to the nucleotide sequence of ACCTACACCATGAAGGAGGTGCTGTTCTACCTGGGCCAGTACATCATGACCAAGCGCCTGTACGACGAGAAGCAGCAGCACATCGTGTACTGCTCCAACGACCTGCTGGGCGACCTGTTCGGCGTGCCCTCCTTCTCCGTGAAGGAGCACCGCAAaATCTACACCATGATCTACCGCAACCTGGTGGTGGTGAACCAGCAGGAGTCCTCCGACTCCGGCACCTCCGTGTCC (SEQ ID NO: 29). a nucleotide sequence having 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity with: ACCTACACCATGAAGGAGATCATCTTCTACATCGGCCAGTACATCATGACCAAGCGCCTGTACGACGAGAAGCAGCAGCACATCGTGTACTGCTCCAACGACCTGCTGGGCGACGTGTTCGGCGTGCCCTCCTTCTCCGTGAAGGAGCAC 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 CGCAAGATCTACGCCATGATCTACCGCAACCTGGTGGCCGTGTCCCAGCAGGACTCCGGCACCTCCCTGTCC (SEQ ID NO: 30); ATCTACACCATGAAGGAGATCATCTTCTACATCGGCCAGTACATCATGACCAAGC At least 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95% of the nucleotide sequence of GCCTGTACGACGAGAAGCAGCAGCACATCGTGTACTGCTCCAACGACCTGCTGGGCGACGTGTTCGGCGTGCCCTCCTTCTCCGTGAAGGAGCACCGCAAGATCTACGCCATGATCTACCGCAACCTGGTGGTGGTGTCCCAGCAGGACTCCGGCACCTCCCCCTCC (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 IYTMKEIIFYIGQYIMTKRLY The peptide may comprise a nucleic acid sequence encoding an epitope of MDM2 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 DEKQQHIVYCSNDLLGDVFGVPSFSVKEHRKIYAMIYRNLVVVSQQDSGTSPS (SEQ ID NO: 34);

[0203] In an illustrative example, the composition comprises:ATGGCGGTACCCATGCAACTGTCCTGCTCTAGACAGAACGGCACCTGGCCCCGCGAGGTGCTGCTGGTGCTGTCCGTGAACTCCTCCGTGTTCCTGCACCTACAGGCCCTGGGCATCCCCCTGCACCTGGCCTACAACTCCTCCCTGGTGACCTTCCAGGAGCCCCCCGGCGTGAACACCACCGAGCTGAGATCCACCGGTGGAGTGCCAGTGCAGGGCTCCAAGTACGCTGCCGACCGCAACCACTACCGCCGCTACCCACGCCGTCGCGGCCCACCCCGCAACTACCAGCAGAACACGCGTGGCCTCAATGCGCACGGCGCAGCGCAGATGCAGCCCATGCACCGCTACGACGTGAGCGCCCTGCAGTACAACTCCATGACCAGCTCGCAGACCTACATGAACGGCTCGCCCACCTACAGCATGTCCTACTCGCAGCAGGGCACCCCTGGCATGGCTCTTGGCTCCATGGGTTCGGTGAGATCCCAATTGAGGTCACTGAAGGAAAGGAATCCATTGAAAATCTTCCCATCCAAACGTATCTTACGAAGACACAAGAGAGATTGGGTGGTTGCTCCAATATCTGTCCCTGAAAATGGCAAGGGTCCCTTCCCACAGAGACTGAATCAGCTCAAGTCTAATAAAGATAGAGACACCAAGATTTTCTACAGCATCACGGGGCCGGGTGCAGACAGCCCACCTGAGGGTGTCTTCGCTGTAGAGAAGGAGACAAGATCCGCCGGCGAAACCTACACCATGAAGGAGGTGCTGTTCTACCTGGGCCAGTACATCATGACCAAGCGCCTGTACGACGAGAAGCAGCAGCACATCGTGTACTGCTCCAACGACCTGCTGGGCGACCTGTTCGGCGTGCCCTCCTTCTCCGTGAAGGAGCACCGCAAAATCTACACCATGATCTACCGCAACCTGGTGGTGGTGAACCAGCAGGAGTCCTCCGACTCCGGCACCTCCGTGTCCAGATCTTAG (SEQ ID NO: 35) nucleotides,38) 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 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 PLKIFPSKRILRRHKRDWVVAPISVPENGKGPFPQRLNQLKSNKDRDTKIFYSITGPGADSPPEGVFAVEKETRSAGETYTMKEVLFYLGQYIMTKRLYDEKQQHIVYCSNDLLGDLFGVPSFSVKEHRKIYTMIYRNLVVVNQQESSDSGTSVSRS (SEQ ID NO: 39);MAVPMTVFMRLNIISPDLSGCTSKGLVLPAVLGITFGAFLIGALLTAALWYIYSHTRSPSKREPVVAVAAPASSESSSTNHSIGSTQSTPCSTSSMATGGVPVQGSKYAADRNHYR RYPRRRGPPRNYQQNTRGLNAHGAAQMQPMHRYDVSALQYNSMTSSQTYMNGSPTYSMSYSQQGTPGMALGSMGSVRSQLLKIFPSKRILRRHKRDWVVAPISVPENGKGPFPQRL Nucleotides 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 NQLKSNKDRDTKIFYSITGPGADSPPEGVFAVEKETRSAGETYTMKEVLFYLGQYIMTKRLYDEKQQHIVYCSNDLLGDLFGVPSFSVKEHRKIYTMIYRNLVVVNQQESSDSGTSVSRS (SEQ ID NO: 40) Sequence;MAVPMTVSMRLNIVSPDLSGKGLVLPSVLGITFGAFLIGALLTAALWYIYSHTRGPSKREPVVAVAAPASSESSSTNHSIGSTQSTPCSTSSMATGGVPVQGSKYAADRNHYRRYPRRRGPPRNYQQNTRGLNAHGAAQMQPMHRYDVSALQYNSMTSSQTYMNGSPTYSMSYSQQGTPGMALGSMGSVRSQLAMHSPPTRILRRRKREWVMPPIFVPENGKGPFPQRL 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 NQLKSNKDRGTKIFYSITGPGADSPPEGVFTIEKESRSAGETYTMKEIIFYIGQYIMTKRLYDEKQQHIVYCSNDLLGDVFGVPSFSVKEHRKIYAMIYRNLVAVSQQDSGTSLSRS (SEQ ID NO: 41);orMAVPMTVSMRLNIVSPDLSGKGLVLPSVLGITFGAFLIGALLTAALWYIYSHTRAPSKREPVVAVAAPASSESSSTNHSIGSTQSTPCSTSSMATGGVPVQGSKYAADRNHYRRYPRRRGPPRNYQQNTRGLNAHGAAQMQPMHRYDVSALQYNSMTSSQTYMNGSPTYSMSYSQQGTPGMALGSMGSVRSQLVMNSPPSRILRRRKREWVMPPISVPENGKGPFPQRLNQLKSNKDRGTKLFYSITGPGAD The nucleic acid sequence may include a nucleic acid sequence encoding a fusion peptide of five epitopes selected from the group consisting of nucleotide sequences 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 SPPEGVFTIEKETRSAGEIYTMKEIIFYIGQYIMTKRLYDEKQQHIVYCSNDLLGDVFGVPSFSVKEHRKIYAMIYRNLVVVSQQDSGTSPSRS (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 including a first nucleotide sequence encoding a first epitope of a first antigen, where the first epitope is a portion of a peptide selected from IGFBP-2, HER-2, and IGF-1R, and the first nucleotide sequence is located on the plasmid. In some examples, the composition can include a first plasmid including 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, where 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, the composition is ATGCTGCCGAGAGTGGGCTGCCCCGCGCTGCCGCTGCCGCCGCCGCCGCTGCTGCCGCTGCTGCCGCTGCTGCTGCTGCTACTGGGCGCGAGTGGCGGCGGCGGCGGGGCGCGCGGAG GTGCTGTTCCGCTGCCCGCCCTGCACACCCGAGCGCCTGGCCGCCTGCGGGCCCCCGCCGGTTGCGCCGCCCGCCGCGGTGGCCGCAGTGGCCGGAGGCGCCCGCATGCCATGCGCGGAGCTCGTCCGGGAGC CGGGCTGCGGCTGCTGCTCGGTGTGCGCCCGGCTGGAGGGCGAGGCGTGCGGCGTCTACACCCCGCGCTGCGGCCAGGGGCTGCGCTGCTATCCCCACCCGGGCTCCGAGCTGCCCCTGCAGGCGCTGGTCATGGGCGAGGGCACTTGTGAGAAGCGCCGGGACGCCGAGTATGGCGCCAGCCCGGAGCAGGTTGCAGACAATGGCGATGACCACTCAGAAGGAGGCCTGGTGGAG (SEQ ID NO: 43) a nucleotide sequence having 0%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity;ATGCTGCCCCGCCTGGGCGGCCCCGCCCTGCCCCTGCTGCTGCCCTCCCTGCTGCTGCTGCTGCTGCTGGGCGCCGGCGGCTGCGGCCCCGGCGTGCGCGCCGAGGTGCTGTTCCGCTGCCCCCCCTGCACCCCCGAGCGCCTGGCCGCCTGCGGCCCCCCCCCCGACGCCCCCTGCGCCGAGCTGGTGC at least 50%, 60%, or at least 50% to the nucleotide sequence of GCGAGCCCGGCTGCGGCTGCTGCTCCGTGTGCGCCCGCCAGGAGGGCGAGGCCTGCGGCGTGTACATCCCCCGCTGCGCCCAGACCCTGCGCTGCTACCCCAACCCCGGCTCCGAGCTGCCCCTGAAGGCCCTGGTGACCGGCGCCGGCACCTGCGAGAAGCGCCGCGTGGGCACCACCCCCCAGCAGGTGGCCGACTCCGACGACGACCACTCCGAGGGCGGCCTGGTGGAG (SEQ ID NO: 44); A nucleotide sequence having 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity;ATGCTGCCCCGCCTGGGCGGCCCCGCCCTGCCCCTGCTGCTGCCCTCCCTGCTGCTGCTGCTGCTGCTGGGCGCCGGCGGCTGCGGCCCCGGCGTGCGCGCCGAGGTGCTGTTCCGCTGCCCCCCCTGCACCCCCGAGCGCCTGGCCGCCTGCGGCCCCCCCCCCGACGCCCCCTGCGCCGAGCTGGTGCGCG At least 50%, 60%, 70% to the nucleotide sequence of AGCCCGGCTGCGGCTGCTGCTCCGTGTGCGCCCGCCAGGAGGGCGAGGCCTGCGGCGTGTACATCCCCCGCTGCGCCCAGACCCTGCGCTGCTACCCCAACCCCGGCTCCGAGCTGCCCCTGAAGGCCCTGGTGACCGGCGCCGGCACCTGCGAGAAGCGCCGCGTGGGCACCACCCCCCAGCAGGTGGCCGACTCCGAGGACGACCACTCCGAGGGCGGCCTGGTGGAG (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 the sequence: TMRRLLQETELVEPLTPSGAMPNQAQMRILKETELRKVKVLGSGAFGTVYKGIWIPDGENVKIPVAIKVLRENTSPKANKEILDEAYVMAGVGSPYVSRLLGICLTSTVQLVTQLMPYGCLLDHVRENRGRLGSQDLLNWCMQIAKGMSYL At least 50% of the amino acid sequence of EDVRLVHRDLAARNVLVKSPNHVKITDFGLARLLDIDETEYHADGGKVPIKWMALESILRRRFTHQSDVWSYGVTVWELMTFGAKPYDGIPAREIPDLLEKGERLPQPPICTIDVYMIMVKCWMIDSECRPRFRELVSEFSRMARDPQRFVVIQNEDLAPGAGGMVHHRHRSSSPLPAARPAGATLERPKTLSPGKNGVVKDVFAFGGAVENPEYL (SEQ ID NO: 60) , 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; TMRRLLQETELVEPLTPSGAVPNQAQMRILKETELRKLKVLGSGAFGTVYKGIWIPDGENVKIPVAIKVLRENTSPKANKEILDEAYVMAGVGSPYVSRLLGICLTSTVQLVTQLMPYGCLLDHVREHRGRLGSQDLLNWCVQIAKGMSYLE At least 50% to the amino acid sequence of EVRLVHRDLAARNVLVKSPNHVKITDFGLARLLDIDETEYHADGGKVPIKWMALESILRRRFTHQSDVWSYGVTVWELMTFGAKPYDGIPAREIPDLLEKGERLPQPPICTIDVYMIMVKCWMIDSECRPRFRELVSEFSRMARDPQRFVVIQNEDLALGTGSTAHRRHRSSSPPPPIRPAGATLERPKTLSPGKNGVVKDVFAFGGAVENPEYL (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 TMRRLLQETELVEPLTPSGAMPNQAQMRILKETELRKVKVLGSGAFGTVYKGIWIPDGENVKIPVAIKVLRENTSPKANKEILDEAYVMAGVGSPYVSRLLGICLTSTVQLVTQLMPYGCLLDHVREHRGRLGSQDLLNW CVQIAKGMSYLEDVRLVHRDLAARNVLVKSPNHVKITDFGLARLLDIDETEYHADGGKVPIKWMALESILRRRFTHQSDVWSYGVTVWELMTFGAKPYDGIPAREIPDLLEKGERLPQPPICTIDVYMIMVKCWMIDSECRPRFRELVSEFSRMARDPQRFVVIQNEDLTPGTGSTAHRRHRSSSPLPPVRPAGATLERPKTLSPGKNGVVKDVFAFGGAVENPEYL(SEQ ID NO: At least 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100%, 101%, 102%, 103%, 104%, 105%, 106%, 107%, 108%, 109%, 110%, 111%, 112%, 113 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 (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 (SEQ ID NO: 80);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 (SEQ ID NO: 81);

[0205] In some examples, the composition can include a first and a second epitope independently selected from CD105, Yb-1, SOX-2, CDH3, or MDM2. In some examples, the composition can include a third epitope, wherein 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 a third and a fourth epitope, wherein 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 a third, fourth, and fifth epitope, wherein 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 a first and a second epitope independently selected from IGFBP2, HER-2, or IGF-1R. In some examples, the composition can include a third epitope, wherein the first, second, and third epitopes are independently selected from IGFBP2, HER-2, or IGF-1R.

[0207] In some cases, the composition can be administered to a subject. In some cases, the subject is in need of administration of the composition. In some cases, the composition is effective in inducing an immune response in the subject. In some cases, the composition is effective in eliminating a large number of cells associated with breast cancer in the subject. In some cases, 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 characteristics, pre-neoplastic 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 cases, the first and second nucleic acid sequences are purified to at least 70% purity. In some cases, the first and second nucleic acid sequences are located on a first plasmid and separated by a linker nucleic acid sequence. In some cases, the first nucleic acid sequence is adjacent to the second nucleic acid sequence on the first plasmid.

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

[0212] In some examples, the subject is selected from a human with breast cancer, a mouse with breast cancer, or a rat with 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 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.

[0214] In some examples, the composition includes 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 HIF-1α peptide, and the first nucleotide sequence is located on the plasmid. In other examples, the composition includes 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 a HIF-1α peptide, and the first nucleotide sequence and the second nucleotide sequence are located on 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 differ from those listed herein. In some examples, nucleic acid sequences that are greater than 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100%, or greater than 50%, homologous to the nucleic acid sequences disclosed herein can be used in the compositions described herein.

[0216] The compositions described herein can, in some examples, include a composition that includes a first epitope of a first antigen expressed by a cell associated with breast cancer and a second epitope of a second antigen expressed by a cell associated with breast cancer.

[0217] In some examples, the composition can include at least a first epitope of a first antigen, where 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 include at least a first epitope of a first antigen and at least a second epitope of a second antigen, where 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 peptide CD105 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 EARMLNASIVASFVELPL (SEQ ID NO: 6); or at least to the amino acid sequence of QNGTWPREVLLVLSVNSSVFLHLQALGIPLHLAYNSSLVTFQEPPGVNTTEL (SEQ ID NO: 1). an amino acid sequence having 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 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity; 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).In some examples, at least a 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 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the amino acid sequence of GVPVQGSKYAADRNHYRRYPRRRGPPRNYQQN (SEQ ID NO: 16); 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). In some examples, at least a first epitope of the 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 a first epitope of the peptide CDH3 is at least 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, 100%, 101%, 102%, 103%, 104%, 105%, 106%, 107%, 108%, 109%, 1109%, 1110, 112%, 113%, 114%, 115%, 116%, 117%, 118%, 119%, 120%, 121%, 122%, 123%, 124%, 125%, 126%, 127%, 128%, 129%, 130%, 131%, 132%, 133%, 134%, 135%, 136%, 137%, 138%, 139%, 140%, 141%, 142%, 143%, 144%, 145%, 146%, 147%, 148%, 149%, 150%, 151%, 152%, 153%, 154%, 155%, 156%, 157%, 158%, 159%, 160%, 161%, 162%, 163%, 164%, 165%, 166%, 167%, 168%, 169%, 170%, 171%, 172%, 173%, 174%, 175%, 176%, 177%, 178%, 179%, 180%, 181%, 182%, 183%, 184%, 185%, 18 an amino acid sequence having 99% or 100% sequence identity; 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).In some examples, at least a first epitope of peptide MDM-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 TYTMKEVLFYLGQYIMTKRLYDEKQQHIVYCSNDLLGDLFGVPSFSVKEHRKIYTMIYRNLVVVNQQESSDSGTSV (SEQ ID NO: 32); TYTMKEIIFYIGQYIMTKRLYDEKQQHIVYCSNDLLGDVFGVPSFSVKEHRKIYAMIYRNLVAVSQQDSGTSLS (SEQ ID NO: 33). 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).

[0218] In some examples, the compositions described herein may comprise: MAVPMQLSCSRQNGTWPREVLLVLSVNSSVFLHLQALGIPLHLAYNSSLVTFQEPPGVNTTELRSTGGVPVQGSKYAADRNHYRRYPRRRGPPRNYQQNTRGLNAHGAAQMQPMHRYDVSALQYNSMTSSQTYMNGSPTYSMSYSQQGTPGMALGSMGSVRSQLRSLKERNPLKIFPSKRILRRHKRDWVVAPISVPENGKG 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 PFPQRLNQLKSNKDRDTKIFYSITGPGADSPPEGVFAVEKETRSAGETYTMKEVLFYLGQYIMTKRLYDEKQQHIVYCSNDLLGDLFGVPSFSVKEHRKIYTMIYRNLVVVNQQESSDSGTSVSRS (SEQ ID NO: 39); MAVPMTVFMRLNIISPDLSGCTSKGLVLPAVLGITFGAFLIGALLTAALWYIYSHTRSPSKREPVVAVAAPASSESSSTNHSIGSTQSTPCSTSSMATGGVPVQGSKYAADRN HYRRYPRRRGPPRNYQQNTRGLNAHGAAQMQPMHRYDVSALQYNSMTSSQTYMNGSPTYSMSYSQQGTPGMALGSMGSVRSQLLKIFPSKRILRRHKRDWVVAPISVPENGKG 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 PFPQRLNQLKSNKDRDTKIFYSITGPGADSPPEGVFAVEKETRSAGETYTMKEVLFYLGQYIMTKRLYDEKQQHIVYCSNDLLGDLFGVPSFSVKEHRKIYTMIYRNLVVVNQQESSDSGTSVSRS (SEQ ID NO: 40);MAVPMTVSMRLNIVSPDLSGKGLVLPSVLGITFGAFLIGALLTAALWYIYSHTRGPSKREPVVAVAAPASSESSSTNHSIGSTQSTPCSTSSMATGGVPVQGSKYAADRNHYRRYP RRRGPPRNYQQNTRGLNAHGAAQMQPMHRYDVSALQYNSMTSSQTYMNGSPTYMSYSQQGTPGMALGSMGSVRSQLAMHSPPTRILRRRKREWVMPPIFVPENGKGPFPQRLNQLK 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 SNKDRGTKIFYSITGPGADSPPEGVFTIEKESRSAGETYTMKEIIFYIGQYIMTKRLYDEKQQHIVYCSNDLLGDVFGVPSFSVKEHRKIYAMIYRNLVAVSQQDSGTSLSRS (SEQ ID NO: 41); or MAVPMTVSMRLNIVSPD LSGKGLVLPSVLGITFGAFLIGALLTAALWYIYSHTRAPSKREPVVAVAAPASSESSSTNHSIGSTQSTPCSTSSMATGGVPVQGSKYAADRNHYRRYPRRRGPPRNYQQNTRGLN AHGAAQMQPMHRYDVSALQYNSMTSSQTYMNGSPTYSMSYSQQGTPGMALGSMGSVRSQLVMNSPPSRILRRRKREWVMPPISVPENGKGPFPQRLNQLKSNKDRGTKLFYSITGPG The amino acid sequence of a fusion peptide of five 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 ADSPPEGVFTIEKETRSAGEIYTMKEIIFYIGQYIMTKRLYDEKQQHIVYCSNDLLGDVFGVPSFSVKEHRKIYAMIYRNLVVVSQQDSGTSPSRS (SEQ ID NO: 42);

[0219] The compositions described herein can include compositions 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 IGFBP-2, HER-2, or IGF-1R, and the first nucleotide sequence is located on the plasmid. In some examples, the compositions 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, 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 a first epitope of the peptide IGFBP-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 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% sequence identity to the amino acid sequence of LGLEEPKKLRPPPAR (SEQ ID NO: 48). , 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 RDAEYGASPEQVADNGDDHSEGGLVE (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 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: MLPRLGGPALPLLLPSLLLLLLLGAGGCGPGVRAEVLFRCPPCTPERLAACGPPPDAPCAELVREPGCGCCSVCARQEGEACGVYIPRCAQTLRCYPNPGSELPLKALVTGAGTCEKRRVGATPQQVADSEDDHSEGGLVE (SEQ ID NO: 56). In some examples, at least a first epitope of the peptide HER-2 has at least 50% similarity to the amino acid sequence of TMRRLLQETELVEPLTPSGAMPNQAQMRILKETELRKVKVLGSGAFGTVYKGIWIPDGENVKIPVAIKVLRENTSPKANKEILDEAYVMAGVGSPYVSRLLGICLTSTVQLVTQLMPYGCLLDHVRENRGRLGSQDLLNWCMQIAKGMSYLEDVRLVHRDLAARNVLVKSPNHVKITDFGLARLLDIDETEYHADGGKVPIKWMALESILRRRFTHQSDVWSYGVTVWELMTFGAKPYDGIPAREIPDLLEKGERLPQPPICTIDVYMIMVKCWMIDSECRPRFRELVSEFSRMARDPQRFVVIQNEDLAPGAGGMVHHRHRSSSPLPAARPAGATLERPKTLSPGKNGVVKDVFAFGGAVENPEYL (SEQ ID NO: 60); a nucleotide sequence encoding an amino acid sequence with 0%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity; TMRRLLQETELVEPLTPSGAVPNQAQMRILKETELRKLKVLGSGAFGTVYKGIWIPDGENVKIPVAIKVLRENTSPKANKEILDEAYVMAGVGSPYVSRLLGICLTSTVQLVTQLMPYGCLLDHVREHRGRLGSQDLLNWCVQI AKGMSYLEEVRLVHRDLAARNVLVKSPNHVKITDFGLARLLDIDETEYHADGGKVPIKWMALESILRRRFTHQSDVWSYGVTVWELMTFGAKPYDGIPAREIPDLLEKGERLPQPPICTIDVYMIMVKCWMIDSECRPRFRELVSEFSRMARDPQRFVVIQNEDLALGTGSTAHRRHRSSSPPPPIRPAGATLERPKTLSPGKNGVVKDVFAFGGAVENPEYL (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 TMRRLLQETELVEPLTPSGAMPNQAQMRILKETELRKVKVLGSGAFGTVYKGIWIPDGENVKIPVAIKVLRENTSPKANKEILDEAYVMAGVGSPYVSRLLGICLTSTVQLVTQLMPYGCLLDHVREHRGRLGSQDLLNWCV At least 50% to the amino acid sequence of QIAKGMSYLEDVRLVHRDLAARNVLVKSPNHVKITDFGLARLLDIDETEYHADGGKVPIKWMALESILRRRFTHQSDVWSYGVTVWELMTFGAKPYDGIPAREIPDLLEKGERLPQPPICTIDVYMIMVKCWMIDSECRPRFRELVSEFSRMARDPQRFVVIQNEDLTPGTGSTAHRRHRSSSPLPPVRPAGATLERPKTLSPGKNGVVKDVFAFGGAVENPEYL (SEQ ID NO: 62), In some examples, the nucleic acid sequence encoding the peptide IGF-IR epitope is selected from the group consisting of: 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 to the amino acid sequence of DYRSYRFPKLTVITE (SEQ ID NO: 66); 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 VVTGYVKIRHSHALV (SEQ ID NO: 68); 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 FFYVQAKTGYENFIH (SEQ ID NO: 69); 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 LIIALPVAVLLIVGG (SEQ ID NO: 70); 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 LVIMLYVFHRKRNNS (SEQ ID NO: 70); 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 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); 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).

[0220] The compositions described herein include 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 (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 (SEQ ID NO: 80);or (SEQ ID NO: 81).

[0221] In some examples, the composition comprises a first and a second epitope independently selected from CD105, Yb-1, SOX-2, CDH3, or MDM2. In some examples, the composition further comprises a third epitope, wherein 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 a third and a fourth epitope, wherein 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 a third, fourth, and fifth epitope, wherein 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 a first and a second epitope independently selected from IGFBP2, HER-2, or IGF-1R. In some examples, the composition further comprises a third epitope, wherein the first, second, and third epitopes are independently selected from IGFBP2, HER-2, or IGF-1R.

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

[0224] In some examples, the composition comprises 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); 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).

[0225] In some examples, the composition comprises 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); or 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 cases, the plasmid backbone can encode one short epitope.In other cases, 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 illustrative 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 cases, the plasmid can encode one extended epitope. In other cases, the compositions can encode more than one extended epitope. For example, the plasmid can encode 2 extended epitopes, 3 extended epitopes, 4 extended epitopes, 5 extended epitopes, 6 extended epitopes, 7 extended epitopes, 8 extended epitopes, 9 extended epitopes, 10 extended epitopes, 11 extended epitopes, 12 extended epitopes, 13 extended epitopes, 14 extended epitopes, 15 extended epitopes, 16 extended epitopes, 17 extended epitopes, 18 extended epitopes, 19 extended epitopes, 20 extended epitopes, or more than 20 extended epitopes. In an illustrative example, the plasmid encodes 4 or fewer extended epitopes.

[0228] The breast cancer vaccine compositions described herein can contain short epitopes and extended epitopes in a single plasmid backbone.In some cases, the plasmid can contain one short epitope.In other cases, the plasmid compositions described herein can contain more than one short epitope.For example, the plasmid compositions 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 ... It is possible to encode 19 extended epitopes, 20 extended epitopes or more than 20 extended epitopes.

[0230] Plasmids for compositions containing more than one sequence encoding an epitope can contain a spacer between each epitope sequence. In some cases, short epitope sequences can be coded in tandem without a spacer. In some cases, extended epitope sequences can be coded in tandem without a spacer. In some cases, short epitope sequences can be coded in tandem using a spacer. In some cases, extended epitope sequences can be coded in tandem using a spacer.

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

[0232] In an illustrative example, a nucleic acid coding sequence for an antigenic epitope peptide can be assembled with a Kozak consensus translation initiation sequence, a stop codon, and a cloning site in a 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 ligases), can be used to generate nucleic acids (e.g., DNA fragments) and insert nucleic acid fragments into a plasmid vector backbone.

[0233] In some cases, the plasmid can contain a nucleic acid sequence encoding at least one tag.In some cases, the tag can be translated into a peptide.Any nucleic acid sequence for tag known to those skilled in the art can be used with the plasmid 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, or a histidine tag with 6 histidine residues, etc.The expression of tag in a subject can be determined using any suitable technique known to those skilled in the art.

[0234] In some examples, the plasmid may 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.

[0235] In some embodiments, the composition may be a multi-antigen breast cancer vaccine. For example, a multi-antigen breast cancer vaccine can contain multiple antigens. In some cases, the expression of one antigen may affect the expression of a different antigen. In some cases, the expression of more than one antigen may affect the expression of a different antigen. In some cases, the expression of one antigen may affect the expression of more than one different antigen. In some cases, the expression of one antigen may not affect the expression of a different antigen. In some cases, the expression of more than one antigen may not affect the expression of a different antigen. In some cases, the expression of one antigen may not affect the expression of more than one different antigen. For example, antigenic competition may limit the immunogenicity of a multi-antigen vaccine. Any technique known to those skilled in the art can be used to determine whether the immune response elicited after administration of a multi-antigen vaccine is comparable in magnitude to each antigen in a single-antigen vaccine. For example, ELISPOT (e.g., for IFNγ secretion) can determine the magnitude of the immune response. In some cases, 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 nucleic acid-based vaccines, including plasmids 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 the expression of the epitope in a subject after administration. For example, the encoded epitope from the nucleic acid construct can allow the immune response to at least one epitope to be synchronized, amplified, attenuated, suppressed, or eliminated in a specific set of proteins (e.g., self-proteins).

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

[0238] In some cases, peptide or nucleic acid constructs can be optimized for protein or plasmid-based vaccination to induce, amplify or synchronize TH1 immune response.In some cases, epitope can be extended TH1 epitope.In other cases, peptide or nucleic acid constructs can be optimized for protein or plasmid-based vaccination to suppress, attenuate or eliminate pathological response in subjects (e.g., humans or animals) that need it.

[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., a vaccine).

[0240] Any suitable pharmaceutical preparation known to those skilled in the art for the expression of nucleic acid sequences. Mido backbones (eg, vectors) 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 direct the expression of a specific nucleotide sequence in an appropriate 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, meaning that at least one of its components is heterologous to at least one of the other components. 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 may be a plasmid, which may be useful for transfecting cells with a nucleic acid encoding a polypeptide, and the transformed host cells may be cultured and maintained under conditions that allow expression of the polypeptide.

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

[0244] The plasmid can further include an initiation codon, which can be upstream of the coding sequence, and a termination codon, which can be downstream of the coding sequence. The initiation and termination 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 human actin, human myosin, human hemoglobin, human muscle creatine, or a viral enhancer, such as one derived from CMV, FMDV, RSV, or EBV. Polynucleotide function enhancement is described in U.S. Patent Nos. 5,593,972, 5,962,428, and WO94 / 016737.

[0245] The plasmid may also contain a mammalian origin of replication to maintain the plasmid extrachromosomally and produce multiple copies of the plasmid in the cell. The plasmid may be pVAXI, pCEP4, or pREP4 from Invitrogen (San Diego, CA).

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

[0247] In some cases, commercially available plasmid backbones can be used. For example, plasmid pUMVC3 can be used. In some cases, commercially available plasmid backbones can be modified, mutated, genetically engineered, or cloned before use. In other cases, 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 Saccharomyces cerevisiae strains 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 the target cell by integration into the cell genome or can exist extrachromosomally (e.g., an autonomously replicating plasmid with an origin of replication). Exemplary vectors include pVAX, pcDNA3.0, or provax, or any other expression vector that can express DNA encoding an antigen and cause the cell to translate the sequence into an antigen that is 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. LECs can be any linear DNA lacking any phosphate backbone. The DNA can encode one or more polypeptides disclosed herein. LECs can contain promoters, introns, stop codons, and / or polyadenylation signals. Polypeptide expression can be controlled by a promoter. LECs cannot contain any antibiotic resistance genes and / or phosphate backbones. LECs cannot contain other nucleic acid sequences unrelated to polypeptide expression.

[0251] 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 that can express DNA encoding an antigen and allow the cell to translate the sequence into an antigen that is recognized by the immune system.

[0252] Prior to insertion of the nucleic acid sequence of at least one epitope, the plasmid backbone may be selected from the group consisting of 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 0kB, approximately 5.2kB, approximately 5.4kB, approximately 5.6kB, approximately 5.8kB, approximately 6.0kB, approximately 6.2kB, approximately 6.4kB, approximately 6.6kB, approximately 6.8kB, approximately 7.0kB, approximately 7.2kB, approximately 7.4kB, approximately 7.6kB, approximately 7.8kB, approximately 8.0kB, approximately 8.2kB, approximately 8.4kB, approximately 8.6kB, approximately 8.8kB, approximately 9.0kB, approximately 9.2kB, approximately 9.4kB, approximately 9.6kB, approximately 9.8kB, approximately 10.0kB, approximately 10. 2kB, approximately 10.4kB, approximately 10.6kB, approximately 10.8kB, approximately 11.0kB, approximately 11.2kB, approximately 11.4kB, approximately 11.6kB, approximately 11.8kB, approximately 12.0kB, approximately 12.2kB, approximately 12.4kB, 12.6kB, 12.8kB, 13.0kB, 13.2kB, 13.4kB, 13.6kB, 13.8kB, 14kB, 14.5kB, 15kB, 15.5kB, 16kB, 1 The length of the plasmid may be less than 6.5 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 an illustrative example, prior to 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, three, four, five, six, seven, eight, nine, ten, eleven, twelve, thirteen, fourteen, fifteen, sixteen, seventeen, eighteen, nineteen, twenty, or more than twenty plasmids.

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

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

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

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

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

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

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

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

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

[0263] In some examples, the composition can include a nucleic acid encoding one or more epitopes from the proteins survivin, HIF-1A, IGFBP2, and IGF-1R. In some cases, the plasmid can include a nucleic acid sequence encoding a polypeptide comprising 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 comprising 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 comprising 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 comprising at least 90% sequence identity to IGFBP-2 (SEQ ID NO: 54). Optionally, the plasmid can include a nucleic acid sequence encoding a polypeptide comprising at least 95% sequence identity to IGFBP-2 (SEQ ID NO: 54). Optionally, the plasmid can include a nucleic acid sequence encoding a polypeptide comprising 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 comprising 100% sequence identity to IGFBP-2 (SEQ ID NO: 54). In some cases, the plasmid can include a nucleic acid sequence encoding a polypeptide comprising 100% sequence identity to IGFBP-2 (SEQ ID NO: 54).

[0264] In some examples, the plasmid can comprise a nucleic acid sequence encoding a polypeptide comprising at least 70% sequence identity over at least 100 to at least 163 contiguous amino acids to IGFBP-2 (SEQ ID NO: 54). In some examples, the plasmid can comprise a nucleic acid sequence encoding a polypeptide comprising at least 70% sequence identity over 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 to IGFBP-2 (SEQ ID NO: 54).

[0265] In some examples, the plasmid can include a nucleic acid sequence encoding a polypeptide comprising at least 80% sequence identity over at least 100 to at least 163 contiguous amino acids to IGFBP-2 (SEQ ID NO: 54). In some examples, the plasmid can include a nucleic acid sequence encoding a polypeptide comprising at least 80% sequence identity over 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 to IGFBP-2 (SEQ ID NO: 54).

[0266] In some examples, the plasmid can comprise a nucleic acid sequence encoding a polypeptide comprising at least 90% sequence identity over at least 100 to at least 163 contiguous amino acids to IGFBP-2 (SEQ ID NO: 54). In some examples, the plasmid can comprise a nucleic acid sequence encoding a polypeptide comprising at least 90% sequence identity over 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 to IGFBP-2 (SEQ ID NO: 54).

[0267] In some examples, the plasmid can comprise a nucleic acid sequence encoding a polypeptide comprising at least 95% sequence identity over at least 100 to at least 163 contiguous amino acids to IGFBP-2 (SEQ ID NO: 54). In some examples, the plasmid can comprise a nucleic acid sequence encoding a polypeptide comprising at least 95% sequence identity over 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 to IGFBP-2 (SEQ ID NO: 54).

[0268] In some examples, the plasmid can include a nucleic acid sequence encoding a polypeptide that comprises 100% sequence identity for at least 100 to at least 163 contiguous amino acids to IGFBP-2 (SEQ ID NO: 54). In some examples, the plasmid can include a nucleic acid sequence encoding a polypeptide that comprises 100% sequence identity for 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 to 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 for at least 100 to at least 163 contiguous amino acids to 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 for 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 to IGFBP-2 (SEQ ID NO: 54).

[0270] In some cases, the plasmid can comprise a nucleic acid sequence comprising at least 50% sequence identity to IGFBP-2 (SEQ ID NO: 43). In some cases, the plasmid can comprise a nucleic acid sequence comprising 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 cases, the plasmid can comprise a nucleic acid sequence comprising at least 70% sequence identity to IGFBP-2 (SEQ ID NO: 43). In some cases, the plasmid can comprise a nucleic acid sequence comprising at least 80% sequence identity to IGFBP-2 (SEQ ID NO: 43). In some cases, the plasmid can comprise a nucleic acid sequence comprising at least 90% sequence identity to IGFBP-2 (SEQ ID NO: 43). Optionally, the plasmid can comprise a nucleic acid sequence comprising at least 95% sequence identity to IGFBP-2 (SEQ ID NO: 43). In some cases, the plasmid can include a nucleic acid sequence that includes at least 99% sequence identity to IGFBP-2 (SEQ ID NO: 43). In some cases, the plasmid can include a nucleic acid sequence that includes 100% sequence identity to IGFBP-2 (SEQ ID NO: 43). In some cases, the plasmid can include a nucleic acid sequence that consists of 100% sequence identity to IGFBP-2 (SEQ ID NO: 43).

[0271] In some cases, the plasmid can comprise a nucleic acid sequence encoding a polypeptide comprising at least 70% sequence identity to survivin (SEQ ID NO: 85). In some cases, the plasmid can comprise a nucleic acid sequence encoding a polypeptide comprising 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 cases, the plasmid can comprise a nucleic acid sequence encoding a polypeptide comprising at least 80% sequence identity to survivin (SEQ ID NO: 85). In some cases, the plasmid can comprise a nucleic acid sequence encoding a polypeptide comprising at least 90% sequence identity to survivin (SEQ ID NO: 85). Optionally, the plasmid can comprise a nucleic acid sequence encoding a polypeptide comprising at least 95% sequence identity to survivin (SEQ ID NO: 85). Optionally, the plasmid can comprise a nucleic acid sequence encoding a polypeptide comprising at least 99% sequence identity to survivin (SEQ ID NO: 85). In some cases, the plasmid can include a nucleic acid sequence encoding a polypeptide comprising 100% sequence identity to survivin (SEQ ID NO: 85). In some cases, the plasmid can include 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 comprising at least 70% sequence identity over at least 10 to at least 38 consecutive amino acids to survivin (SEQ ID NO: 85). In some examples, the plasmid can comprise a nucleic acid sequence encoding a polypeptide comprising at least 70% sequence identity over at least 12 to at least 35, at least 15 to at least 30, or at least 20 to at least 25 consecutive amino acids to survivin (SEQ ID NO: 85). In some examples, the plasmid can comprise a nucleic acid sequence encoding a polypeptide comprising at least 70% sequence identity over at least 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 32, 34, or 36 consecutive amino acids to survivin (SEQ ID NO: 85).

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

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

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

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

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

[0278] In some cases, the plasmid can comprise a nucleic acid sequence comprising at least 50% sequence identity to survivin (SEQ ID NO: 86). In some cases, the plasmid can comprise a nucleic acid sequence comprising 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 cases, the plasmid can comprise a nucleic acid sequence comprising at least 70% sequence identity to survivin (SEQ ID NO: 86). In some cases, the plasmid can comprise a nucleic acid sequence comprising at least 80% sequence identity to survivin (SEQ ID NO: 86). In some cases, the plasmid can comprise a nucleic acid sequence comprising at least 90% sequence identity to survivin (SEQ ID NO: 86). Optionally, the plasmid can comprise a nucleic acid sequence comprising at least 95% sequence identity to survivin (SEQ ID NO: 86). Optionally, the plasmid can comprise a nucleic acid sequence comprising at least 99% sequence identity to survivin (SEQ ID NO: 86). In some cases, the plasmid can include a nucleic acid sequence that comprises 100% sequence identity to survivin (SEQ ID NO: 86). In some cases, the plasmid can include a nucleic acid sequence that consists 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 comprising 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 comprising 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 comprising 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 comprising at least 90% sequence identity to HIF-1A (SEQ ID NO: 87). Optionally, the plasmid can include a nucleic acid sequence encoding a polypeptide comprising at least 95% sequence identity to HIF-1A (SEQ ID NO: 87). Optionally, the plasmid can include a nucleic acid sequence encoding a polypeptide comprising 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 comprising 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 comprise a nucleic acid sequence encoding a polypeptide comprising at least 70% sequence identity over at least 40 to at least 89 consecutive amino acids of HIF-1A (SEQ ID NO: 87). In some examples, the plasmid can comprise a nucleic acid sequence encoding a polypeptide comprising at least 70% sequence identity over 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 consecutive amino acids of HIF-1A (SEQ ID NO: 87). In some examples, the plasmid can comprise a nucleic acid sequence encoding a polypeptide comprising at least 70% sequence identity over 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 consecutive amino acids of HIF-1A (SEQ ID NO: 87).

[0281] In some examples, the plasmid can comprise a nucleic acid sequence encoding a polypeptide comprising at least 80% sequence identity for at least 40 to at least 89 consecutive amino acids of HIF-1A (SEQ ID NO: 87). In some examples, the plasmid can comprise a nucleic acid sequence encoding a polypeptide comprising at least 80% sequence identity for 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 consecutive amino acids of HIF-1A (SEQ ID NO: 87). In some examples, the plasmid can comprise a nucleic acid sequence encoding a polypeptide comprising at least 80% sequence identity for 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 consecutive amino acids of HIF-1A (SEQ ID NO: 87).

[0282] In some examples, the plasmid can comprise a nucleic acid sequence encoding a polypeptide comprising at least 90% sequence identity for at least 40 to at least 89 consecutive amino acids of HIF-1A (SEQ ID NO: 87). In some examples, the plasmid can comprise a nucleic acid sequence encoding a polypeptide comprising at least 90% sequence identity for 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 consecutive amino acids of HIF-1A (SEQ ID NO: 87). In some examples, the plasmid can comprise a nucleic acid sequence encoding a polypeptide comprising at least 90% sequence identity for 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 consecutive amino acids of HIF-1A (SEQ ID NO: 87).

[0283] In some examples, the plasmid can comprise a nucleic acid sequence encoding a polypeptide comprising at least 95% sequence identity for at least 40 to at least 89 consecutive amino acids of HIF-1A (SEQ ID NO: 87). In some examples, the plasmid can comprise a nucleic acid sequence encoding a polypeptide comprising at least 95% sequence identity for 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 consecutive amino acids of HIF-1A (SEQ ID NO: 87). In some examples, the plasmid can comprise a nucleic acid sequence encoding a polypeptide comprising at least 95% sequence identity for 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 consecutive amino acids of HIF-1A (SEQ ID NO: 87).

[0284] In some examples, the plasmid can comprise a nucleic acid sequence encoding a polypeptide comprising 100% sequence identity for at least 40 to at least 89 consecutive amino acids to HIF-1A (SEQ ID NO: 87). In some examples, the plasmid can comprise a nucleic acid sequence encoding a polypeptide comprising 100% sequence identity for 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 consecutive amino acids to HIF-1A (SEQ ID NO: 87). In some examples, the plasmid can comprise a nucleic acid sequence encoding a polypeptide comprising 100% sequence identity for 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 consecutive amino acids to HIF-1A (SEQ ID NO: 87).

[0285] In some examples, the plasmid can comprise 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 comprise 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 comprise 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 comprising at least 50% sequence identity to HIF-1A (SEQ ID NO: 88). In some cases, the plasmid can comprise a nucleic acid sequence comprising 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 cases, the plasmid can comprise a nucleic acid sequence comprising at least 70% sequence identity to HIF-1A (SEQ ID NO: 88). In some cases, the plasmid can comprise a nucleic acid sequence comprising at least 80% sequence identity to HIF-1A (SEQ ID NO: 88). In some cases, the plasmid can comprise a nucleic acid sequence comprising at least 90% sequence identity to HIF-1A (SEQ ID NO: 88). Optionally, the plasmid can comprise a nucleic acid sequence comprising at least 95% sequence identity to HIF-1A (SEQ ID NO: 88). In some cases, the plasmid can include a nucleic acid sequence that includes at least 99% sequence identity to HIF-1A (SEQ ID NO: 88). In some cases, the plasmid can include a nucleic acid sequence that includes 100% sequence identity to HIF-1A (SEQ ID NO: 88). In some cases, the plasmid can include a nucleic acid sequence that consists of 100% sequence identity to HIF-1A (SEQ ID NO: 88).

[0287] In some cases, the plasmid can include a nucleic acid sequence encoding a polypeptide comprising at least 70% sequence identity to IGF-IR (SEQ ID NO: 73). In some cases, the plasmid can include a nucleic acid sequence encoding a polypeptide comprising 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 include a nucleic acid sequence encoding a polypeptide comprising at least 80% sequence identity to IGF-IR (SEQ ID NO: 73). In some examples, the plasmid can include a nucleic acid sequence encoding a polypeptide comprising at least 90% sequence identity to IGF-IR (SEQ ID NO: 73). Optionally, the plasmid can include a nucleic acid sequence encoding a polypeptide comprising at least 95% sequence identity to IGF-IR (SEQ ID NO: 73). Optionally, the plasmid can include a nucleic acid sequence encoding a polypeptide comprising at least 99% sequence identity to IGF-IR (SEQ ID NO: 73). In some cases, the plasmid can include a nucleic acid sequence encoding a polypeptide comprising 100% sequence identity to IGF-IR (SEQ ID NO: 73). In some cases, the plasmid can include a nucleic acid sequence encoding a polypeptide consisting of 100% sequence identity to IGF-IR (SEQ ID NO: 73).

[0288] In some examples, the plasmid can include a nucleic acid sequence encoding a polypeptide comprising at least 70% sequence identity over 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 comprising at least 70% sequence identity over 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 comprising at least 80% sequence identity for at least 50 to at least 104 contiguous amino acids to IGF-1R (SEQ ID NO: 73). In some examples, the plasmid can include a nucleic acid sequence encoding a polypeptide comprising at least 80% sequence identity for 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 to IGF-1R (SEQ ID NO: 73).

[0290] In some examples, the plasmid can include a nucleic acid sequence encoding a polypeptide comprising at least 90% sequence identity for 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 comprising at least 90% sequence identity for 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 comprising at least 95% sequence identity over 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 comprising at least 95% sequence identity over 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 that comprises 100% sequence identity for at least 50 to at least 104 contiguous amino acids to IGF-1R (SEQ ID NO: 73). In some examples, the plasmid can include a nucleic acid sequence encoding a polypeptide that comprises 100% sequence identity for 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 to 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 for at least 50 to at least 104 contiguous amino acids to 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 for 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 to IGF-1R (SEQ ID NO: 73).

[0294] In some cases, the plasmid can comprise a nucleic acid sequence comprising at least 50% sequence identity to IGF-IR (SEQ ID NO: 63). In some cases, the plasmid can comprise a nucleic acid sequence comprising 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 cases, the plasmid can comprise a nucleic acid sequence comprising at least 70% sequence identity to IGF-IR (SEQ ID NO: 63). In some cases, the plasmid can comprise a nucleic acid sequence comprising at least 80% sequence identity to IGF-IR (SEQ ID NO: 63). In some cases, the plasmid can comprise a nucleic acid sequence comprising at least 90% sequence identity to IGF-IR (SEQ ID NO: 63). Optionally, the plasmid can comprise a nucleic acid sequence comprising at least 95% sequence identity to IGF-IR (SEQ ID NO: 63). In some cases, the plasmid can include a nucleic acid sequence that includes at least 99% sequence identity to IGF-IR (SEQ ID NO: 63). In some cases, the plasmid can include a nucleic acid sequence that includes 100% sequence identity to IGF-IR (SEQ ID NO: 63). In some cases, the plasmid can include a nucleic acid sequence that consists of 100% sequence identity to IGF-IR (SEQ ID NO: 63).

[0295] Optionally, the isolated and purified plasmid can comprise at least one nucleic acid sequence encoding a polypeptide comprising at least 70% sequence identity to an epitope sequence selected from SEQ ID NOs: 54, 73, 85, and 87. The plasmid can comprise at least one nucleic acid sequence encoding a polypeptide comprising 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 comprise at least one nucleic acid 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 plasmid can comprise at least one nucleic acid 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 plasmid can comprise at least one nucleic acid 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 plasmid can comprise at least one nucleic acid 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 plasmid can comprise at least one nucleic acid sequence encoding a polypeptide comprising 100% sequence identity to an epitope sequence selected from SEQ ID NOs: 54, 73, 85, and 87. The plasmid can comprise 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] Optionally, the isolated and purified plasmid can comprise at least four nucleic acid sequences, each of the four nucleic acid sequences encoding a polypeptide comprising at least 70% sequence identity to an epitope sequence selected from SEQ ID NOs: 54, 73, 85, and 87. The plasmid can comprise at least four nucleic acid sequences, each of the four nucleic acid sequences encoding a polypeptide comprising 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 comprise at least four nucleic acid sequences, each of the four nucleic acid sequences encoding a polypeptide comprising at least 80% sequence identity to an epitope sequence selected from SEQ ID NOs: 54, 73, 85, and 87. The plasmid can comprise at least four nucleic acid sequences, each of the four nucleic acid sequences encoding a polypeptide comprising at least 90% sequence identity to an epitope sequence selected from SEQ ID NOs: 54, 73, 85, and 87. The plasmid can comprise at least four nucleic acid sequences, each of the four nucleic acid sequences encoding a polypeptide comprising at least 95% sequence identity to an epitope sequence selected from SEQ ID NOs: 54, 73, 85, and 87. The plasmid can comprise at least four nucleic acid sequences, each of the four nucleic acid sequences encoding a polypeptide comprising at least 99% sequence identity to an epitope sequence selected from SEQ ID NOs: 54, 73, 85, and 87. The plasmid can comprise at least four nucleic acid sequences, each of the four nucleic acid sequences encoding a polypeptide comprising 100% sequence identity to an epitope sequence selected from SEQ ID NOs: 54, 73, 85, and 87. The plasmid can include at least 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 cases, the at least four nucleic acid sequences independently encode polypeptides to an epitope sequence selected from SEQ ID NOs: 54, 73, 85, and 87.In other cases, the at least four nucleic acid sequences encode different polypeptides directed to an epitope sequence selected from SEQ ID NOs: 54, 73, 85, and 87.

[0297] Optionally, the plasmid can comprise four nucleic acid sequences, each of the four nucleic acid sequences encoding a polypeptide comprising at least 70% sequence identity to an epitope sequence selected from SEQ ID NOs: 54, 73, 85, and 87. The plasmid can comprise four nucleic acid sequences, each of the four nucleic acid sequences encoding a polypeptide comprising 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 comprise four nucleic acid sequences, each of the four nucleic acid sequences encoding a polypeptide comprising at least 80% sequence identity to an epitope sequence selected from SEQ ID NOs: 54, 73, 85, and 87. The plasmid can comprise four nucleic acid sequences, each of which encodes a polypeptide comprising at least 90% sequence identity to an epitope sequence selected from SEQ ID NOs: 54, 73, 85, and 87. The plasmid can comprise four nucleic acid sequences, each of which encodes a polypeptide comprising at least 95% sequence identity to an epitope sequence selected from SEQ ID NOs: 54, 73, 85, and 87. The plasmid can comprise four nucleic acid sequences, each of which encodes a polypeptide comprising at least 99% sequence identity to an epitope sequence selected from SEQ ID NOs: 54, 73, 85, and 87. The plasmid can comprise four nucleic acid sequences, each of which encodes a polypeptide comprising 100% sequence identity to an epitope sequence selected from SEQ ID NOs: 54, 73, 85, and 87. The plasmid can include 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 cases, the four nucleic acid sequences independently encode polypeptides for epitope sequences selected from SEQ ID NOs: 54, 73, 85, and 87. In other cases, the four nucleic acid sequences encode different polypeptides for epitope sequences selected from SEQ ID NOs: 54, 73, 85, and 87.

[0298] In some cases, the plasmid can comprise a nucleic acid sequence encoding a polypeptide comprising at least 70% sequence identity to SEQ ID NO:89. In some examples, the plasmid can comprise a nucleic acid sequence encoding a polypeptide comprising 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 comprise a nucleic acid sequence encoding a polypeptide comprising at least 80% sequence identity to SEQ ID NO:89. The plasmid can comprise a nucleic acid sequence encoding a polypeptide comprising at least 90% sequence identity to SEQ ID NO:89. The plasmid can comprise a nucleic acid sequence encoding a polypeptide comprising at least 95% sequence identity to SEQ ID NO:89. The plasmid can comprise a nucleic acid sequence encoding a polypeptide comprising at least 99% sequence identity to SEQ ID NO:89. The plasmid can comprise a nucleic acid sequence encoding a polypeptide comprising 100% sequence identity to SEQ ID NO:89.

[0299] In some cases, the plasmid can comprise a nucleic acid sequence comprising at least 70% sequence identity to SEQ ID NO:90. In some examples, the plasmid can comprise a nucleic acid sequence comprising 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 comprising at least 80% sequence identity to SEQ ID NO:90. The plasmid can comprise a nucleic acid sequence comprising at least 90% sequence identity to SEQ ID NO:90. The plasmid can comprise a nucleic acid sequence comprising at least 95% sequence identity to SEQ ID NO:90. The plasmid can comprise a nucleic acid sequence comprising at least 99% sequence identity to SEQ ID NO:90. The plasmid can comprise a nucleic acid sequence comprising 100% sequence identity to SEQ ID NO:90.

[0300] In some cases, the plasmid can comprise a nucleic acid sequence comprising at least 70% sequence identity to SEQ ID NO:91. In some examples, the plasmid can comprise a nucleic acid sequence comprising 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 comprising at least 80% sequence identity to SEQ ID NO:91. The plasmid can comprise a nucleic acid sequence comprising at least 90% sequence identity to SEQ ID NO:91. The plasmid can comprise a nucleic acid sequence comprising at least 95% sequence identity to SEQ ID NO:91. The plasmid can comprise a nucleic acid sequence comprising at least 99% sequence identity to SEQ ID NO:91. The plasmid can comprise a nucleic acid sequence comprising 100% sequence identity to SEQ ID NO:91.

[0301] In some cases, a plasmid containing more than one epitope sequence can include a spacer between each epitope sequence. In some cases, epitope sequences can be encoded in tandem without a spacer. In some cases, epitope sequences can be encoded in tandem using a spacer. In some cases, the spacer can include a sequence encoding about 1 to about 50, about 3 to about 40, about 5 to about 35, or 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 cases, the plasmid can contain a nucleic acid sequence encoding at least one tag.In some cases, the tag can be translated into a peptide.Any nucleic acid sequence for tag known to those skilled in the art can be used with the plasmid 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, or a histidine tag with 6 histidine residues, etc.The expression of tag in a subject can be determined using any suitable technique known to those skilled in the art.

[0303] In some examples, the plasmid may 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 may be a multi-antigen breast cancer vaccine or a multi-antigen ovarian cancer vaccine. For example, a multi-antigen breast cancer vaccine or a multi-antigen ovarian cancer vaccine can contain multiple antigens. In some cases, the expression of one antigen may affect the expression of a different antigen. In some cases, the expression of more than one antigen may affect the expression of a different antigen. In some cases, the expression of one antigen may affect the expression of more than one different antigen. In some cases, the expression of one antigen may not affect the expression of a different antigen. In some cases, the expression of more than one antigen may not affect the expression of a different antigen. In some cases, the expression of one antigen may not affect the expression of more than one different antigen. For example, antigenic competition may limit the immunogenicity of a multi-antigen vaccine. Any technique known to those skilled in the art can be used to determine whether the immune response elicited after administration of a multi-antigen vaccine is comparable in magnitude to each antigen in a single-antigen vaccine. For example, ELISPOT (e.g., for IFNγ secretion) can determine the magnitude of the immune response. In some cases, ELISPOT can detect rodent, non-human primate, or human peptides. In some cases, a multi-antigen breast or ovarian cancer vaccine can include multiple epitopes 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), which is the genome or chromosome in which the nucleic acid molecule is naturally found. As such, "isolated" does not necessarily reflect the extent to which the nucleic acid molecule has been purified, but indicates that the molecule does not contain 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 that contains a gene is not a fragment of a chromosome containing such a gene, but rather contains the coding and regulatory regions associated with the gene, but is free of additional genes that are naturally found on the same chromosome. An isolated nucleic acid molecule can also contain a designated nucleic acid sequence flanked by additional nucleic acids (i.e., at the 5' and / or 3' ends of the sequence) that are not normally adjacent to the designated nucleic acid sequence in nature (i.e., heterologous sequences).

[0306] Isolated nucleic acid molecules can contain both genomic and cDNA DNA, RNA, or hybrids in which the nucleic acid can contain combinations 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 or recombinant methods. The phrase "nucleic acid molecule" primarily refers to the physical nucleic acid molecule, and the phrase "nucleic acid sequence" primarily refers to the sequence of nucleotides in a nucleic acid molecule, although the two terms can be used interchangeably, particularly with respect to nucleic acid molecules or nucleic acid sequences capable of encoding proteins or protein domains.

[0307] A nucleic acid molecule can refer to at least two nucleotides covalently linked together. The nucleic acids described herein can contain phosphodiester bonds, although in some instances, as outlined below (e.g., in the construction of probes such as primers and labeled probes), they may contain phosphodiester bonds, e.g., phosphoramide bonds (Beaucage et al., Tetrahedron 49(10):1925 (1993) and references therein; Letsinger, J. Org. Chem. 35:3800 (1970); Sprinzl et al., Eur. J. Biochem. 81:579 (1977); Letsinger et al., Nucl. Acids Res. 14:3487 (1986); Sawai et al., Chem. Lett. 805 (1984); Letsinger et al., J. Am. Chem. Soc. 110:4470 (1988); and Pauwels et al., Chemica Scripta 26:141 (1986)), phosphorothioates (Mag et al., Nucleic Acids Res. 19:1437 (1991); and U.S. Pat. No. 5,644,048), phosphorodithioates (Briu et al., J. Am. Chem. Soc. 111:2321 (1989)), O-methylphosphoroamidite linkages (see Eckstein, Oligonucleotides and Analogues: A Practical Approach, Oxford University Press), and peptide nucleic acid (also referred to herein as "PNA") backbones and linkages (Egholm, J. Am. Chem. Soc. 114:1895 (1992); Meier et al., Chem. Int. Ed. Engl. 31:1008 (1992); Nielsen, Nature 365:566 (1993); Carlsson et al., Nature 105:107 (1994), all of which are incorporated by reference. 380:207 (1996) Other analog nucleic acids include locked nucleic acids (also referred to herein as "LNAs"), Koshkin et al., J. Am. Chem. Soc. 120:13. 252-3 (1998); normal 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," edited by Y.S. Sanghui and P. Dan Cook; Mesmaeker et al., Bioorganic & Medicinal Chem. Lett. 4:395 (1994) 4; Jeffs et al., J. Biomolecular NMR 34:17 (1994); Tetrahedron Lett. 37:743 (1996)), and U.S. Pat. Nos. 5,235,033 and 5,034,506 and Chapters 6 and 7, ASC Symposium Series Vol. 580, "Carbohydrate Modifications in Antisense Research," Y.S. Sanghui and bicyclic structures containing non-ribose backbones, including those described in P. Dan Cook, ed. Nucleic acids containing one or more carbocyclic sugars are also included within the definition of nucleic acid (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 acids" are also included within the definition of nucleic acid analog. 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 expressly incorporated herein by reference. These modifications of the ribose-phosphate backbone can be made to increase the stability and half-life of such molecules in physiological environments. For example, PNA:DNA and LNA-DNA hybrids can exhibit greater stability and 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 and single-stranded sequence. 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 any combination of bases, including uracil, adenine, thymine, cytosine, guanine, inosine, xanthine hypoxanthine, isocytosine, isoguanine, etc.

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

[0309] A recombinant nucleic acid molecule includes any nucleic acid sequence, typically a heterologous sequence, operably linked to an isolated nucleic acid molecule encoding a fusion protein of the present invention, capable of enabling recombinant production of the fusion protein and delivering the nucleic acid molecule into a host cell according to the present invention. Such vectors can contain nucleic acid sequences not naturally found adjacent to the isolated nucleic acid molecule to be inserted into the vector. Vectors can be either RNA or DNA, prokaryotic or eukaryotic, and are preferably viruses or plasmids in the present invention. Recombinant vectors can be used in the cloning, sequencing, and / or other manipulation of nucleic acid molecules and in the delivery of such molecules (e.g., in DNA compositions or viral vector-based compositions). Recombinant vectors are preferably used in the expression of nucleic acid molecules and can also be referred to as expression vectors. Preferred recombinant vectors are capable of expression in transfected host cells.

[0310] In the recombinant molecules of the invention, the nucleic acid molecule is operably linked to an expression vector that is compatible with the host cell and contains regulatory sequences, such as transcriptional control sequences, translational control sequences, origins of replication, and other regulatory sequences that control expression of the nucleic acid molecule of the invention. In particular, recombinant molecules of the invention comprise a nucleic acid molecule operably linked to one or more expression control sequences. The phrase "operably linked" refers to the linkage of a nucleic acid molecule to an expression control sequence in a manner such that the molecule is expressed upon transfection (i.e., transformation, transduction, or transfection) into a host cell. Pharmaceutical Composition

[0311] The immunogenic compositions of the present disclosure are preferably formulated as vaccines for in vivo administration to subjects to confer antibody titers that exceed the threshold for antibody prevalence to each antigenic component in an acceptable percentage of subjects. Antigens with associated antibody titers above which a subject is considered to have seroconverted to the antigen are well known, and such titers are published by organizations such as the WHO. Preferably, greater than 80% of a statistically significant sample of subjects will seroconvert, more preferably greater than 90%, even more preferably greater than 93%, and most preferably 96-100%. Adjuvants

[0312] The immunogenic composition of the present disclosure is preferably adjuvanted. Adjuvants can be used to enhance the immune response (humoral and / or cellular) induced in vaccinated patients. In some cases, adjuvants can induce a TH1-type response. In other cases, adjuvants can induce a TH2-type response. In contrast to TH2-type responses, which can be characterized by the production of cytokines such as IL-4, IL-5, and IL-10, TH1-type responses can be characterized by the production of cytokines such as IFN-γ.

[0313] Adjuvants can include stimulatory molecules such as cytokines, including, but not limited to, CCL20, alpha-interferon (IFN-a), beta-interferon (IFN-β), gamma-interferon, platelet-derived growth factor (PDGF), tumor necrosis factor (TNFa), tumor necrosis factor (TNFp), granulocyte-macrophage colony-stimulating factor (GM-CSF), epidermal growth factor (EGF), cutaneous T cell-attracting chemokine (CTACK), epithelial thymus-expressed chemokine (TECK), mucosal-associated epithelial chemokine (MEC), IL-12, IL-15, IL-28, MHC, CD80, CD86, IL-1, IL-2, IL-4, IL-5, IL-6, IL-10, IL-18, MCP-1, MIP-1a, MIP-1-, IL-8, L-selectin, P-selectin, E-selectin, CD34, and GlyCAM. -1, MadCAM-1, LFA-1, VLA-1, Mac-1, pl50.95, PECAM, ICAM-1, ICAM-2, ICAM-3, CD2, LFA-3, M-CSF, G-CSF, mutant form of IL-18, CD40, CD40L, vascular growth factor, fibroblast growth factor, IL-7, nerve growth factor, vascular endothelial growth factor, Fas, TNF receptor, Fit, Apo-1, p55, WSL-1, DR3, TRAMP, Apo-3, AIR, LARD, NGRF, DR4, DRS, KILLER, TRAIL-R2, TRICK2, DR6, Caspase ICE, Fos, c-jun, Sp-1, Ap-1, Ap-2, p38, p65Rel, MyD88, IRAK, TRAF6, IkB, Inactive NIK, SAP K, SAP-I, JNK, interferon response genes, NFkB, Bax, TRAIL, TRAILrec, TRAILrecDRC5, TRAIL-R3, TRAIL-R4, RANK, RANK LIGAND, Ox40, Ox40 LIGAND, NKG2D, MICA, MICB, NKG2A, NKG2B, NKG2C, NKG2E, NKG2F, TAPI, and TAP2. In some cases, the adjuvant is GM-CSF.

[0314] Additional adjuvants include MCP-1, MIP-1a, MIP-1p, IL-8, RANTES, L-selectin, P-selectin, E-selectin, CD34, GlyCAM-1, MadCAM-1, LFA-1, VLA-1, Mac-1, pl50.95, PECAM, ICAM-1, ICAM-2, ICAM-3, CD2, LFA-3, M-CSF, G-CSF, IL-4, mutant forms of IL-18, CD40, CD40L, vascular growth factor, fibroblast growth factor, and IFN-γ. Cell growth factor, IL-7, IL-22, nerve growth factor, vascular endothelial growth factor, Fas, TNF receptor, Fit, Apo-1, p55, WSL-1, DR3, TRAMP, Apo-3, AIR, LARD, NGRF, DR4, DR5, KILLER, TRAIL-R2, TRICK2, DR6, caspase ICE, Fos, c-jun, Sp-1, Ap-1, Ap-2, p38, p65Rel, MyD88, IRAK, TRAF6, IkB, inactive NIK, SAP K, SAP-1, JNK, interferon response gene, NFkB, Bax, TRAIL, TRAILrec, TRAILrecDRC5, TRAIL-R3, TRAIL-R4, RANK, RANK LIGAND, Ox40, Ox40 These include LIGAND, NKG2D, MICA, MICB, NKG2A, NKG2B, NKG2C, NKG2E, NKG2F, TAP1, TAP2, and functional fragments thereof.

[0315] In some embodiments, the adjuvant can be a modulator of a toll-like receptor. Examples of toll-like receptor modulators include TLR-9 agonists, but are not limited to small molecule modulators of toll-like receptors, such as imiquimod. Other examples of adjuvants that can be used in combination with the vaccines described herein can include, but are not limited to, saponins, CpG ODNs, and the like.

[0316] Optionally, the adjuvant can include aluminum hydroxide gel (alum), aluminum salts such as aluminum phosphate, salts of calcium, iron, or zinc, or can be an insoluble suspension of acylated tyrosine or acylated sugar, cationic or anionic derivatized polysaccharide, or polyphosphazene.

[0317] In some cases, a suitable adjuvant system that promotes a predominantly Th1 response includes monophosphoryl lipid A or a derivative thereof, particularly a combination of 3-O-deacylated monophosphoryl lipid A and monophosphoryl lipid A, preferably 3-O-deacylated monophosphoryl lipid A (3D-MPL), together with an aluminum salt. An enhanced system involves a combination of monophosphoryl lipid A and a saponin derivative, particularly the combination of QS21 and 3D-MPL disclosed in WO 94 / 00153, or the less reactogenic composition in which QS21 is quenched with cholesterol disclosed in WO 96 / 33739. A particularly potent adjuvant formulation involving QS21, 3D-MPL, and tocopherol in an oil-in-water emulsion is described in WO 95 / 17210. The vaccine can additionally include a saponin, more preferably QS21. Formulations may also include an oil-in-water emulsion and tocopherol (WO95 / 17210). Unmethylated CpG-containing oligonucleotides (WO96 / 02555) are also preferential inducers of Th1 responses and are suitable for use in the present disclosure.

[0318] In some cases, aluminum salts are used. Optionally, the polysaccharide conjugate may be unadjuvanted to minimize the level of adjuvants (especially aluminum salts) in the compositions of the present disclosure.

[0319] Optionally, suitable adjuvant systems can include adjuvants or immunostimulants, including, but not limited to, detoxified lipid A and non-toxic derivatives of lipid A from any source, saponins, and other agents capable of stimulating TH1-type responses. Enterobacterial lipopolysaccharide (LPS) is known to be a potent stimulator of the immune system, although its use in adjuvants has been curtailed by its toxic effects. A non-toxic derivative of LPS, monophosphoryl lipid A (MPL), produced by removal of the core carbohydrate group and phosphate from the reducing-end glucosamine, is described in Ribi et al. (1986, Immunology and Immunopharmacology of Bacterial Endotoxins, Plenum Publishing Corp., NY, pp. 407-419).

[0320] A further detoxified version of MPL is derived by removal of the acyl chain from the 3-position of the disaccharide backbone and is called 3-O-deacylated monophosphoryl lipid (Upid) A (3D-MPL), which can be purified and prepared by the methods taught in GB 2122204B, which also discloses the preparation of diphosphoryl lipid A and its 3-O-deacylated variant.

[0321] In some cases, 3D-MPL is in the form of an emulsion with small particle sizes less than 0.2 μm in diameter, and its preparation method is disclosed in WO 94 / 21292. Aqueous formulations containing monophosphoryl lipid A and a surfactant are described in WO 9843670 A2. Bacterial lipopolysaccharide-derived adjuvants to be formulated in the compositions of the present disclosure can be purified and processed from bacterial sources or can be synthesized. For example, purified monophosphoryl lipid A is described in Ribi et al., 1986 (see above), and 3-O-deacylated monophosphoryl or diphosphoryl lipid A from Salmonella sp. is described in GB 2220211 and US 4912094. Other purified and synthetic lipopolysaccharides have been described (Hilgers et al., 1986, Int. ArchAllergy. Immunol 79(4):392-6; Hilgers et al., 1987, Immunology 60(1):141-6; and EP 0549074 B1. A particularly preferred bacterial lipopolysaccharide adjuvant is 3D-MPL.

[0322] Thus, the LPS derivatives that can be used in the present disclosure are immunostimulants that are structurally similar to LPS, MPL, or 3D-MPL. In another aspect of the present disclosure, the LPS derivatives can be acylated monosaccharides that are a small portion of the above-described structure of MPL.

[0323] Saponins are listed in Lacaille-Dubois, M. and Wagner, H. (1996, A review of the biological and pharmacological activities of saponins., Phytomedicine, 2, pp. 363-386). Saponins are steroid or triterpene glycosides widely distributed in the plant and marine animal kingdoms. Saponins are notorious for forming foaming colloidal solutions in water when shaken and for precipitating cholesterol. When saponins are near cell membranes, they induce pore-like structures in the membrane, causing it to rupture. Hemolysis of red blood cells is an example of this phenomenon, a property of certain, but not all, saponins.

[0324] Saponins are known as adjuvants in vaccines for systemic administration. The adjuvant and hemolytic activity of individual saponins has been extensively studied in the art (Lacaille-Dubois and Wagner, supra). For example, Quil A (a plant of the South American tree Quillaja (derived from the bark of Saponaria Molina) and its fractions are disclosed in US 5,057,540 and in "Saponins as vaccine adjuvants", Kensil, CR, CritRev TherDrug Carrier Syst, 1996, 12(1-2):1-55; and EP0362279B1.

[0325] Particulate structures termed immune stimulating complexes (ISCOMS), which contain fractions of Quil A, are hemolytic and have been used in vaccine production (Morein, B., EP010 9942B1; WO96 / 11711; WO96 / 33739). The hemolytic saponins QS21 and QS17 (HPLC-purified fractions of Quil A) have been described as potent systemic adjuvants, and methods for their production are disclosed in U.S. Pat. No. 5,057,540 and EP 0 362 279 B1. Other saponins that have been used in systemic vaccination trials include saponins from other plant species, such as Gypsophila and Saponaria (Bomford et al., Vaccine 10(9):572-577, 1992).

[0326] Enhanced systems involve combinations of non-toxic lipid A derivatives and saponin derivatives, in particular the combination of QS21 and 3D-MPL as disclosed in WO94 / 00153, or less reactogenic compositions in which QS21 is quenched with cholesterol as disclosed in WO96 / 33739.

[0327] Optionally, the adjuvant is selected from a bacterial toxoid, a polyoxypropylene-polyoxyethylene block polymer, an aluminum salt, a liposome, a CpG polymer, an oil-in-water emulsion, or a combination thereof.

[0328] In some cases, adjuvant is oil-in-water emulsion.Oil-in-water emulsion can comprise at least one oil and at least one surfactant, and oil(s) and surfactant(s) are biodegradable (metabolizable) and biocompatible.The oil droplets in emulsion generally have a diameter of less than 5 μ m, and can even have a submicron diameter, and these small sizes can be achieved by microfluidizer to produce stable emulsion.Preferably, the droplets have a size of less than 220 nm, because they can be subjected to filtration sterilization.

[0329] The oils used can include oils derived from animal (e.g., fish) or vegetable sources. Vegetable oil sources can include nuts, seeds, and grains. Peanut oil, soybean oil, coconut oil, and olive oil are the most commonly available examples of nut oils. For example, jojoba oil obtained from jojoba beans can be used. Seed oils include safflower oil, cottonseed oil, sunflower seed oil, sesame seed oil, and the like. The grain group can include corn oil and oils from other grains such as wheat, oat, rye, rice, teff, triticale, and others. 6-10 carbon fatty acid esters of glycerol and 1,2-propanediol do not naturally occur in seed oils but can be prepared by hydrolysis, separation, and esterification of appropriate materials starting from nut and seed oils. Fats and oils derived from mammalian milk can be metabolized and therefore can be used with the vaccines described herein. Procedures for separation, purification, saponification, and other means necessary to obtain pure oils from animal sources are well known in the art. Fish can contain easily recoverable metabolizable oils. For example, whale oils such as cod liver oil, shark liver oil, and spermaceti are examples of some of the fish oils that can be used herein. Many branched-chain oils can be biochemically synthesized in five-carbon isoprene units and can be generally referred to as terpenoids. Shark liver oil contains a branched, unsaturated terpenoid known as squalene, 2,6,10,15,19,23-hexamethyl-2,6,10,14,18,22-tetracosahexaene. Squalane, a saturated analog of squalene, can also be used. Squalene and fish oils containing squalane are readily available from commercial sources or can be obtained by methods known in the art.

[0330] Other useful oils include tocopherols, which can be included in vaccines for elderly patients (e.g., 60 years or older), because vitamin E has been reported to have a positive effect on immune responses in this patient group. Furthermore, tocopherols have antioxidant properties that can help stabilize emulsions. There are various tocopherols (α, β, γ, δ, ε, or ξ), but α is commonly used. An example of α-tocopherol is DL-α-tocopherol. α-tocopherol succinate can be compatible with cancer vaccines and can be a useful preservative as an alternative to mercury compounds.

[0331] Mixtures of oils can be used, for example, squalene and α-tocopherol. Oil contents ranging from 2 to 20% (by volume) can be used.

[0332] Surfactants can be classified by their "HLB" (hydrophilic / lipophilic balance). In some instances, surfactants have an HLB of at least 10, at least 15, and / or at least 16. Surfactants include...

Claims

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 NOs: 1, 16, 20, 26, and 32.

2. The nucleic acid described in 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.

3. The nucleic acid described in 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.

4. The nucleic acid described in claim 1, wherein the amino acid sequence of the fusion peptide is the amino acid sequence of SEQ ID NO:

39.

5. A nucleic acid described in any one of claims 1 to 4, wherein the nucleotide sequence is located in a pUMVC3 plasmid backbone.

6. The nucleic acid described in 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.

7. The nucleic acid described in claim 6, wherein the adjuvant is GM-CSF.

8. The nucleic acid described in claim 7, which, when administered to a subject, is capable of inducing a Th1 immune response.

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