Compositions and methods for treating and/or preventing a variety of diseases and conditions that are amenable to immunotherapy and, in one particular embodiment, compositions and methods for treating and/or preventing cancer in an animal are described. Specifically improvements related to the use o
Compositions and methods for treating and/or preventing a variety of diseases and conditions that are amenable to immunotherapy and, in one particular embodiment, compositions and methods for treating and/or preventing cancer in an animal are described. Specifically improvements related to the use of a yeast-based vaccine comprising a yeast vehicle and an antigen that is selected to elicit an antigen-specific cellular and humoral immune response in an animal, for use in prophylactic and/or therapeutic vaccination and the prevention and/or treatment of a variety of diseases and conditions are disclosed.
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1. A method to reduce tumor burden, increase survival, or inhibit tumor growth in an animal that has cancer, comprising administering a composition to an animal that has a cancer, to reduce tumor burden, increase survival, or inhibit tumor growth in the animal, the composition comprising: a) a yeast
1. A method to reduce tumor burden, increase survival, or inhibit tumor growth in an animal that has cancer, comprising administering a composition to an animal that has a cancer, to reduce tumor burden, increase survival, or inhibit tumor growth in the animal, the composition comprising: a) a yeast vehicle; andb) a fusion protein expressed by the yeast vehicle, the fusion protein comprising: i) an amino acid sequence comprising positions 57-65 of a Ras protein, wherein, as compared to wild-type Ras protein, the glutamine at position 61 is mutated; andii) an amino acid sequence comprising positions 3-165 of a Ras protein, wherein, as compared to wild-type Ras protein, the glycine at position 12 is mutated, and the glutamine at position 61 is mutated;wherein the amino acid sequence of (i) is appended to the N-terminus of the amino acid sequence of (ii). 2. The method of claim 1, wherein the Ras protein comprises the amino acid sequence of SEQ ID NO:5. 3. The method of claim 1, wherein the Ras protein comprises the amino acid sequence of SEQ ID NO:3. 4. The method of claim 1, wherein the fusion protein further comprises a peptide linked to the N-terminus of the amino acid sequence of (i), the peptide consisting of at least two amino acid residues that are heterologous to the amino acid sequence of (i), wherein the peptide stabilizes the expression of the fusion protein in the yeast vehicle or prevents posttranslational modification of the expressed fusion protein; wherein the amino acid residue at position one of the fusion protein is a methionine;wherein the amino acid residue at position two of the fusion protein is not a glycine or a proline;wherein none of the amino acid residues at positions 2-6 of the fusion protein is a methionine; and,wherein none of the amino acid residues at positions 2-5 of the fusion protein is a lysine or an arginine. 5. The method of claim 4, wherein the peptide consists of between two and six amino acid residues which are heterologous to the amino acid sequence of (i). 6. The method of claim 4, wherein the peptide consists of two amino acid residues which are heterologous to the amino acid sequence of (i). 7. The method of claim 1, wherein the fusion protein further comprises a peptide linked to the N-terminus of the amino acid sequence of (i), the peptide consisting of at least two amino acid residues that are heterologous to the amino acid sequence of (i), wherein the peptide stabilizes the expression of the fusion protein in the yeast vehicle or prevents posttranslational modification of the expressed fusion protein, and wherein the first six amino acids of the fusion protein consist of an amino acid sequence of M-X2-X3-X4-X5-X6; wherein M is methionine;wherein X2 is any amino acid except glycine, proline, lysine or arginine;wherein X3 is any amino acid except methionine, lysine or arginine;wherein X4 is any amino acid except methionine, lysine or arginine;wherein X5 is any amino acid except methionine, lysine or arginine; andwherein X6 is any amino acid except methionine. 8. The method of claim 7, wherein X6 is a proline. 9. The method of claim 7, wherein the peptide consists of an amino acid sequence of M-A-D-E-A-P (SEQ ID NO:1). 10. The method of claim 1, wherein the yeast vehicle is selected from: a whole yeast, a yeast spheroplast, a yeast cytoplast, a yeast ghost, or a subcellular yeast membrane extract or fraction thereof. 11. The method of claim 1, wherein the yeast vehicle is selected from: a whole yeast, a yeast spheroplast, a yeast cytoplast, or a yeast ghost. 12. The method of claim 1, wherein the yeast vehicle is a whole yeast or a yeast spheroplast. 13. The method of claim 1, wherein the yeast vehicle is a whole, killed yeast. 14. The method of claim 1, wherein the yeast vehicle is from a yeast selected from: Saccharomyces, Schizosaccharomyces, Kluveromyces, Hansenula, Candida or Pichia. 15. The method of claim 1, wherein the yeast vehicle is from Saccharomyces cerevisiae. 16. The method of claim 1, wherein the composition further comprises a biological response modifier. 17. The method of claim 1, wherein the composition further comprises an immunopotentiator. 18. The method of claim 1, wherein the composition is administered after surgical resection of a tumor from the animal. 19. The method of claim 1, wherein the composition is administered in combination with another therapeutic treatment for cancer. 20. The method of claim 19, wherein the therapeutic treatment for cancer is chemotherapy. 21. The method of claim 19, wherein the therapeutic treatment for cancer is radiation therapy. 22. The method of claim 19, wherein the therapeutic treatment for cancer is surgery. 23. The method of claim 1, wherein the composition is administered to the animal every two weeks. 24. The method of claim 1, wherein the composition is administered to the animal from about one to about 4 times over a time period of from about 1 month to about 6 months. 25. The method of claim 1, wherein the composition is administered by subcutaneous injection. 26. A method to reduce tumor burden, increase survival, or inhibit tumor growth in an animal that has cancer, comprising administering a composition to an animal that has a cancer, to reduce tumor burden, increase survival, or inhibit tumor growth in the animal, the composition comprising: a) a yeast vehicle; andb) a fusion protein expressed by the yeast vehicle, the fusion protein comprising: i) an amino acid sequence comprising positions 57-65 of a Ras protein, wherein, as compared to wild-type Ras protein, the glutamine at position 61 is substituted with a leucine;ii) an amino acid sequence comprising positions 8-16 of a Ras protein, wherein, as compared to wild-type Ras protein, the glycine at position 12 is substituted with a valine; andiii) an amino acid sequence comprising positions 57-65 of a Ras protein, wherein, as compared to wild-type Ras protein, the glutamine at position 61 is substituted with an arginine. 27. The method of claim 26, wherein the Ras protein comprises the amino acid sequence of SEQ ID NO:5. 28. The method of claim 26, wherein the Ras protein comprises the amino acid sequence of SEQ ID NO:3. 29. The method of claim 26, wherein the yeast vehicle is a whole, killed yeast. 30. The method of claim 26, wherein the yeast vehicle is from Saccharomyces cerevisiae. 31. The method of claim 26, wherein the composition is administered after surgical resection of a tumor from the animal. 32. The method of claim 26, wherein the composition is administered in combination with another therapeutic treatment for cancer. 33. The method of claim 32, wherein the therapeutic treatment for cancer is chemotherapy. 34. A method to reduce tumor burden, increase survival, or inhibit tumor growth in an animal that has cancer, comprising administering a composition to an animal that has a cancer, to reduce tumor burden, increase survival, or inhibit tumor growth in the animal, the composition comprising: a) a yeast vehicle; andb) a fusion protein expressed by the yeast vehicle, wherein the fusion protein comprises the amino acid sequence of SEQ ID NO:15. 35. The method of claim 34, wherein the fusion protein consists of the amino acid sequence of SEQ ID NO:15. 36. The method of claim 34, wherein the yeast vehicle is a whole, killed yeast. 37. The method of claim 34, wherein the yeast vehicle is from Saccharomyces cerevisiae. 38. The method of claim 34, wherein the composition is administered after surgical resection of a tumor from the animal. 39. The method of claim 34, wherein the composition is administered in combination with another therapeutic treatment for cancer. 40. The method of claim 39, wherein the therapeutic treatment for cancer is chemotherapy. 41. A method to reduce tumor burden, increase survival, or inhibit tumor growth in an animal that has cancer, comprising administering a composition to an animal that has a cancer, to reduce tumor burden, increase survival, or inhibit tumor growth in the animal, the composition comprising: a) a yeast vehicle; andb) a fusion protein expressed by the yeast vehicle, the fusion protein comprising: i) an amino acid sequence comprising positions 57-65 of a Ras protein, wherein, as compared to wild-type Ras protein, the glutamine at position 61 is substituted with a leucine;ii) an amino acid sequence comprising positions 8-16 of a Ras protein, wherein, as compared to wild-type Ras protein, the glycine at position 12 is substituted with a cysteine; andiii) an amino acid sequence comprising positions 57-65 of a Ras protein, wherein, as compared to wild-type Ras protein, the glutamine at position 61 is substituted with an arginine. 42. The method of claim 41, wherein the Ras protein comprises the amino acid sequence of SEQ ID NO:5. 43. The method of claim 41, wherein the Ras protein comprises the amino acid sequence of SEQ ID NO:3. 44. The method of claim 41, wherein the yeast vehicle is a whole, killed yeast. 45. The method of claim 41, wherein the yeast vehicle is from Saccharomyces cerevisiae. 46. The method of claim 41, wherein the composition is administered after surgical resection of a tumor from the animal. 47. The method of claim 41, wherein the composition is administered in combination with another therapeutic treatment for cancer. 48. The method of claim 47, wherein the therapeutic treatment for cancer is chemotherapy. 49. A method to reduce tumor burden, increase survival, or inhibit tumor growth in an animal that has cancer, comprising administering a composition to an animal that has a cancer, to reduce tumor burden, increase survival, or inhibit tumor growth in the animal, the composition comprising: a) a yeast vehicle; andb) a fusion protein expressed by the yeast vehicle, the fusion protein comprising: i) an amino acid sequence comprising positions 57-65 of a Ras protein, wherein, as compared to wild-type Ras protein, the glutamine at position 61 is substituted with a leucine;ii) an amino acid sequence comprising positions 8-16 of a Ras protein, wherein, as compared to wild-type Ras protein, the glycine at position 12 is substituted with an aspartic acid; andiii) an amino acid sequence comprising positions 57-65 of a Ras protein, wherein, as compared to wild-type Ras protein, the glutamine at position 61 is substituted with an arginine. 50. The method of claim 49, wherein the Ras protein comprises the amino acid sequence of SEQ ID NO:5. 51. The method of claim 49, wherein the Ras protein comprises the amino acid sequence of SEQ ID NO:3. 52. The method of claim 49, wherein the yeast vehicle is a whole, killed yeast. 53. The method of claim 49, wherein the yeast vehicle is from Saccharomyces cerevisiae. 54. The method of claim 49, wherein the composition is administered after surgical resection of a tumor from the animal. 55. The method of claim 49, wherein the composition is administered in combination with another therapeutic treatment for cancer. 56. The method of claim 55, wherein the therapeutic treatment for cancer is chemotherapy. 57. A method to reduce tumor burden, increase survival, or inhibit tumor growth in an animal that has cancer, comprising administering a composition to an animal that has a cancer, to reduce tumor burden, increase survival, or inhibit tumor growth in the animal, the composition comprising: a) a yeast vehicle; andb) a fusion protein expressed by the yeast vehicle, the fusion protein comprising the amino acid sequence of SEQ ID NO:15, except that as compared to SEQ ID NO:15, the valine at position 25 is substituted with a cysteine. 58. The method of claim 57, wherein the yeast vehicle is a whole, killed yeast. 59. The method of claim 57, wherein the yeast vehicle is from Saccharomyces cerevisiae. 60. The method of claim 57, wherein the composition is administered after surgical resection of a tumor from the animal. 61. The method of claim 57, wherein the composition is administered in combination with another therapeutic treatment for cancer. 62. The method of claim 61, wherein the therapeutic treatment for cancer is chemotherapy. 63. The method of claim 61, wherein the therapeutic treatment for cancer is radiation therapy. 64. The method of claim 61, wherein the therapeutic treatment for cancer is surgery. 65. The method of claim 57, wherein the composition is administered to the animal every two weeks. 66. The method of claim 57, wherein the composition is administered by subcutaneous injection. 67. A method to reduce tumor burden, increase survival, or inhibit tumor growth in an animal that has cancer, comprising administering a composition to an animal that has a cancer, to reduce tumor burden, increase survival, or inhibit tumor growth in the animal, the composition comprising: a) a yeast vehicle; andb) a fusion protein expressed by the yeast vehicle, the fusion protein comprising the amino acid sequence of SEQ ID NO:15, except that as compared to SEQ ID NO:15, the valine at position 25 is substituted with an aspartic acid. 68. The method of claim 67, wherein the yeast vehicle is a whole, killed yeast. 69. The method of claim 67, wherein the yeast vehicle is from Saccharomyces cerevisiae. 70. The method of claim 67, wherein the composition is administered after surgical resection of a tumor from the animal. 71. The method of claim 67, wherein the composition is administered in combination with another therapeutic treatment for cancer. 72. The method of claim 71, wherein the therapeutic treatment for cancer is chemotherapy. 73. The method of claim 71, wherein the therapeutic treatment for cancer is radiation therapy. 74. The method of claim 71, wherein the therapeutic treatment for cancer is surgery. 75. The method of claim 67, wherein the composition is administered to the animal every two weeks. 76. The method of claim 67, wherein the composition is administered by subcutaneous injection. 77. The method of claim 1, wherein the Ras protein is K-Ras. 78. The method of claim 26, wherein the Ras protein is K-Ras. 79. The method of claim 41, wherein the Ras protein is K-Ras. 80. The method of claim 49, wherein the Ras protein is K-Ras. 81. The method of claim 34, wherein the composition is administered to the animal every two weeks. 82. The method of claim 34, wherein the composition is administered by subcutaneous injection. 83. The method of claim 39, wherein the therapeutic treatment for cancer is radiation therapy. 84. The method of claim 39, wherein the therapeutic treatment for cancer is surgery. 85. A method to reduce tumor burden, increase survival, or inhibit tumor growth in an animal that has cancer, comprising administering a composition to an animal that has a cancer, to reduce tumor burden, increase survival, or inhibit tumor growth in the animal, the composition comprising: a) a yeast vehicle; andb) a fusion protein expressed by the yeast vehicle, the fusion protein comprising: i) an amino acid sequence comprising positions 57-65 of a Ras protein, wherein, as compared to wild-type Ras protein, the glutamine at position 61 is substituted with a leucine; andii) an amino acid sequence comprising positions 3-165 of a Ras protein, wherein, as compared to wild-type Ras protein, the glycine at position 12 is substituted with a valine, and the glutamine at position 61 is substituted with an arginine;wherein the amino acid sequence of (i) is appended to the N-terminus of the amino acid sequence of (ii). 86. The method of claim 85, wherein the Ras protein is K-Ras. 87. The method of claim 85, wherein the Ras protein comprises the amino acid sequence of SEQ ID NO:5. 88. The method of claim 85, wherein the Ras protein comprises the amino acid sequence of SEQ ID NO:3. 89. The method of claim 85, wherein the yeast vehicle is a whole, killed yeast. 90. The method of claim 85, wherein the yeast vehicle is from Saccharomyces cerevisiae. 91. The method of claim 85, wherein the composition is administered after surgical resection of a tumor from the animal. 92. The method of claim 85, wherein the composition is administered in combination with another therapeutic treatment for cancer. 93. The method of claim 92, wherein the therapeutic treatment for cancer is chemotherapy. 94. A method to reduce tumor burden, increase survival, or inhibit tumor growth in an animal that has cancer, comprising administering a composition to an animal that has a cancer, to reduce tumor burden, increase survival, or inhibit tumor growth in the animal, the composition comprising: a) a yeast vehicle; andb) a fusion protein expressed by the yeast vehicle, the fusion protein comprising: wherein the fusion protein comprises: i) an amino acid sequence comprising positions 57-65 of a Ras protein, wherein, as compared to wild-type Ras protein, the glutamine at position 61 is substituted with a leucine; andii) an amino acid sequence comprising positions 3-165 of a Ras protein, wherein, as compared to wild-type Ras protein, the glycine at position 12 is substituted with a cysteine, and the glutamine at position 61 is substituted with an arginine;wherein the amino acid sequence of (i) is appended to the N-terminus of the amino acid sequence of (ii). 95. The method of claim 94, wherein the Ras protein is K-Ras. 96. The method of claim 94, wherein the Ras protein comprises the amino acid sequence of SEQ ID NO:5. 97. The method of claim 94, wherein the Ras protein comprises the amino acid sequence of SEQ ID NO:3. 98. The method of claim 94, wherein the yeast vehicle is a whole, killed yeast. 99. The method of claim 94, wherein the yeast vehicle is from Saccharomyces cerevisiae. 100. The method of claim 94, wherein the composition is administered after surgical resection of a tumor from the animal. 101. The method of claim 94, wherein the composition is administered in combination with another therapeutic treatment for cancer. 102. The method of claim 101, wherein the therapeutic treatment for cancer is chemotherapy. 103. A method to reduce tumor burden, increase survival, or inhibit tumor growth in an animal that has cancer, comprising administering a composition to an animal that has a cancer, to reduce tumor burden, increase survival, or inhibit tumor growth in the animal, the composition comprising: a) a yeast vehicle; andb) a fusion protein expressed by the yeast vehicle, the fusion protein comprising: i) an amino acid sequence comprising positions 57-65 of a Ras protein, wherein, as compared to wild-type Ras protein, the glutamine at position 61 is substituted with a leucine; andii) an amino acid sequence comprising positions 3-165 of a Ras protein, wherein, as compared to wild-type Ras protein, the glycine at position 12 is substituted with an aspartic acid, and the glutamine at position 61 is substituted with an arginine;wherein the amino acid sequence of (i) is appended to the N-terminus of the amino acid sequence of (ii). 104. The method of claim 103, wherein the Ras protein is K-Ras. 105. The method of claim 103, wherein the Ras protein comprises the amino acid sequence of SEQ ID NO:5. 106. The method of claim 103, wherein the Ras protein comprises the amino acid sequence of SEQ ID NO:3. 107. The method of claim 103, wherein the yeast vehicle is a whole, killed yeast. 108. The method of claim 103 wherein the yeast vehicle is from Saccharomyces cerevisiae. 109. The method of claim 103, wherein the composition is administered after surgical resection of a tumor from the animal. 110. The method of claim 103, wherein the composition is administered in combination with another therapeutic treatment for cancer. 111. The method of claim 110, wherein the therapeutic treatment for cancer is chemotherapy.
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