Claims for Patent: 6,218,166

Summary for Patent: 6,218,166

Title:	Adjuvant incorporation into antigen carrying cells: compositions and methods
Abstract:	Disclosed are compositions and methods for enhancing the antibody and T cell response to cellular antigens by incorporating an immunopotentiating agent into the cellular membrane or into an intracellular compartment. Such adjuvant-incorporated cell compositions are useful in methods to increase immune responses against antigens, including immunologically cryptic tumor cell antigens, and may be employed to generate useful diagnostic antibodies, to elicit anti-tumor effects in immunized animals, and to significantly prolong survival in animals with cancer.
Inventor(s):	Ravindranath; Mepur H. (Los Angeles, CA), Morton; Donald L. (Malibu, CA)
Assignee:	John Wayne Cancer Institute (Santa Monica, CA)
Application Number:	08/462,106
Patent Claims:	1. A composition comprising a cell that includes an adjuvant non-covalently incorporated into the cell surface membrane or an intracellular compartment of said cell. 2. The composition of claim 1, comprising a cell in which an adjuvant is non-covalently incorporated into the cell surface membrane of said cell. 3. The composition of claim 1, comprising a cell that includes an adjuvant non-covalently incorporated into the cell surface membrane and an adjuvant non-covalently incorporated into an intracellular compartment of said cell. 4. The composition of claim 1, wherein said cell is a human cell. 5. The composition of claim 1, wherein said cell is an erythrocyte. 6. The composition of claim 1, wherein said cell comprises an intracellular antigen. 7. The composition of claim 1, wherein said cell is a tumor cell. 8. The composition of claim 7, wherein said cell is a tumor cell listed in Table 2 or Table 3. 9. The composition of claim 7, wherein said cell is an irradiated tumor cell. 10. The composition of claim 7, wherein said cell is a tumor cell that comprises a tumor-associated intracellular antigen. 11. The composition of claim 7, wherein said cell is a tumor cell that comprises a tumor-associated ganglioside antigen. 12. The composition of claim 7, wherein said cell is a melanoma cell. 13. The composition of claim 12, wherein said cell is a mouse melanoma cell. 14. The composition of claim 13, wherein said cell is the mouse melanoma cell B16. 15. The composition of claim 12, wherein said cell is a human melanoma cell. 16. The composition of claim 15, wherein said cell is the human melanoma cell M27, M18, M14, M111, M22, M7, M102, M108, M16, M104, M109, M25, M24, M10 or M101. 17. The composition of claim 16, wherein said cell is the human melanoma cell M14, M7, M24, M25, M10 or M101. 18. The composition of claim 1, comprising a cell that includes two or more distinct adjuvants non-covalently incorporated into the cell surface membrane or an intracellular compartment of said cell. 19. The composition of claim 1, further comprising a combination of cell types, wherein at least one of which cell types includes an adjuvant non-covalently incorporated into the cell surface membrane or an intracellular compartment of said cell. 20. The composition of claim 1, wherein said adjuvant is an adjuvant listed in Table 1. 21. The composition of claim 20, wherein said adjuvant is lipoteichoic acid (LTA), ribitol technic acid (RTA), glycerol teichoic acid (GTA), hemocyanin from keyhole limpet (KLH), chitin, chitosan, muramyl dipeptide (MDP), threonyl-MDP, a fatty acid derivative of muramyl dipeptide (MTPPE), bacillus Calmette-Guerin (BCG), cell wall skeleton (CWS), trehalose dimycolate, QS21, Quil A or lentinen. 22. The composition of claim 20, wherein said adjuvant is a bacterial superantigen. 23. The composition of claim 20, wherein said adjuvant is of the lipopolysaccharide group of adjuvants. 24. The composition of claim 23, wherein said adjuvant is a detoxified endotoxin. 25. The composition of claim 24, wherein said adjuvant is monophosphoryl lipid A (MPL). 26. The composition of claim 1, comprising a population of cells that includes between about 0.4 ng and about 3.1 ng of cell surface-associated adjuvant per 10.sup.6 cells, wherein said adjuvant is non-covalently incorporated into the cell surface membrane of said cells. 27. The composition of claim 26, comprising a population of cells that includes between about 1.6 ng and about 2.4 ng of cell surface-associated adjuvant per 10.sup.6 cells, wherein said adjuvant is non-covalently incorporated into the cell surface membrane of said cells. 28. The composition of claim 1, dispersed in a pharmacologically acceptable formulation. 29. The composition of claim 1, prepared by a method comprising the steps of: (a) preparing an adjuvant-suspended culture media composition by sonicating an adjuvant with a culture medium; (b) obtaining a cell composition; and (c) admixing said adjuvant-suspended culture media composition and said cell composition under conditions effective and for a period of time suitable to allow non-covalent incorporation of the adjuvant into the cell surface membrane or an intracellular compartment of a cell, thereby preparing said composition. 30. The composition of claim 12, prepared by a method comprising the steps of: (a) preparing an MPL-suspended culture media composition by sonicating MPL with a culture medium; (b) obtaining a melanoma cell composition; and (c) admixing said MPL-suspended culture media composition and said melanoma cell composition under conditions effective and for a period of time suitable to allow non-covalent incorporation of the MPL into the cell surface membrane or an intracellular compartment of a cell, thereby preparing said composition. 31. A method of preparing an adjuvant-cell composition in which an adjuvant is non-covalently incorporated into the cell surface membrane or an intracellular compartment of a cell, comprising admixing an adjuvant composition with a cell composition under conditions effective and for a period of time suitable to allow non-covalent incorporation of the adjuvant into a cell surface membrane or an intracellular compartment of a cell, thereby preparing said adjuvant-cell composition. 32. The method of claim 31, comprising the steps of: (a) preparing an adjuvant-suspended culture media composition; (b) obtaining a cell composition; and (c) admixing said adjuvant-suspended culture media composition and said cell composition under conditions effective and for a period of time suitable to allow non-covalent incorporation of the adjuvant into a cell surface membrane or an intracellular compartment of a cell, thereby preparing said adjuvant-cell composition. 33. The method of claim 32, wherein said adjuvant-suspended culture media composition is prepared by sonication. 34. The method of claim 32, wherein said adjuvant-suspended culture media and said cell composition are admixed at a temperature of between about 10.degree. C. and about 40.degree. C. 35. The method of claim 34, wherein said adjuvant-suspended culture media and said cell composition are admixed at a temperature of about 37.degree. C. 36. A method for stimulating an immune response, comprising administering to an animal an immunologically effective amount of an adjuvant-cell composition comprising a cell that includes an adjuvant non-covalently incorporated into the cell surface membrane or an intracellular compartment of said cell. 37. The method of claim 36, wherein a biological sample is obtained from said animal to provide an antibody. 38. The method of claim 37, wherein a blood sample is obtained from said animal to provide a polyclonal antibody. 39. The method of claim 37, wherein a spleen cell sample is obtained from said animal to provide a monoclonal antibody. 40. The method of claim 36, wherein a biological sample is obtained from said animal to provide an antigen-specific T cell. 41. The method of claim 36, wherein said adjuvant-cell composition comprises an erythrocyte that includes an adjuvant non-covalently incorporated into the erythrocyte cell surface membrane or an intracellular compartment of said erythrocyte. 42. The method of claim 36, wherein said adjuvant-cell composition comprises an irradiated tumor cell that includes an adjuvant non-covalently incorporated into the tumor cell surface or an intracellular compartment of said tumor cell. 43. The method of claim 42, wherein said adjuvant-cell composition comprises an irradiated melanoma cell that includes an adjuvant non-covalently incorporated into the melanoma cell surface or an intracellular compartment of said melanoma cell. 44. The method of claim 36, wherein said adjuvant-cell composition comprises a cell that is obtained from an animal, non-covalently incorporated into said adjuvant in vitro, and then administered to the same animal. 45. The method of claim 36, wherein said adjuvant-cell composition comprises an LTA, RTA, GTA, KLH, chitin, chitosan, MDP, threonyl-MDP, MTPPE, BCG, cell wall skeleton (CWS), trehalose dimycolate, QS21, Quil A or lentinen adjuvant in non-covalent association with the cell surface or an intracellular compartment of said cell. 46. The method of claim 36, wherein said adjuvant-cell composition comprises a bacterial superantigen adjuvant in non-covalent association with the cell surface or an intracellular compartment of said cell. 47. The method of claim 36, wherein said adjuvant-cell composition comprises a detoxified endotoxin adjuvant non-covalently incorporated into the cell surface membrane or an intracellular compartment of said cell. 48. The method of claim 36, wherein said animal is a human subject. 49. The method of claim 36, wherein said adjuvant-cell composition comprises a cell that includes an adjuvant non-covalently incorporated into the cell surface membrane and an adjuvant in non-covalent association with an intracellular compartment of said cell. 50. The method of claim 36, wherein said cell that includes an adjuvant non-covalently incorporated into the cell surface membrane or an intracellular compartment is selected from the group consisting of: an adenocarcinoma cell, adenoma cell, astrocytoma cell, bladder tumor cell, brain tumor cell, Burkitt lymphoma cell, breast carcinoma cell, cervical carcinoma cell, colon carcinoma cell, kidney carcinoma cell, liver carcinoma cell, lung carcinoma cell, ovarian carcinoma cell, pancreatic carcinoma cell, prostate carcinoma cell, rectal carcinoma cell, skin carcinoma cell, stomach carcinoma cell, testis carcinoma cell, thyroid carcinoma cell, chondrosarcoma cell, choriocarcinoma cell, fibroma cell, fibrosarcoma cell, glioblastoma cell, glioma cell, hepatoma cell, histiocytoma cell, leiomyoblastoma cell, leiomyosarcoma cell, leukemia cell, lymphoma cell, liposarcoma cell, mammary tumor cell, medulloblastoma cell, myeloma cell, plasmacytoma cell, neuroblastoma cell, neuroglioma cell, osteogenic sarcoma cell, pancreatic tumor cell, pituitary tumor cell, retinoblastoma cell, rhabdomyosarcoma cell, sarcoma cell, testicular tumor cell, thymoma cell or a Wilms' tumor cell. 51. The method of claim 36, wherein said animal has cancer. 52. The method of claim 48, wherein said human has cancer. 53. A composition comprising a cell that includes an adjuvant that is integrated into the cell surface membrane of the cell, non-covalently incorporated into a cell surface membrane protein of the cell, or that is incorporated into an intracellular compartment of the cell. 54. A method for stimulating an immune response, comprising administering to an animal an immunologically effective amount of an adjuvant-cell composition comprising a cell that includes an adjuvant that is integrated into the cell surface membrane of the cell, non-covalently incorporated into a cell surface membrane protein of the cell, or that is incorporated into an intracellular compartment of the cell. 55. The composition of claim 2, comprising a cell that includes an adjuvant that is integrated into the membrane bilayer at the cell surface of said cell. 56. The composition of claim 2, comprising a cell that includes an adjuvant that is non-covalently incorporated into a membrane protein within the cell surface membrane of said cell. 57. A composition comprising a cell that includes an adjuvant that is integrated into the cell surface membrane of the cell or that is non-covalently incorporated into an intracellular compartment of the cell. 58. A method for stimulating an immune response, comprising administering to an animal an immunologically effective amount of an adjuvant-cell composition comprising a cell that includes an adjuvant that is integrated into the cell surface membrane of the cell or that is non-covalently incorporated into an intracellular compartment of the cell. 59. A composition comprising a cell that includes an adjuvant non-covalently incorporated into the cell surface of the cell and an adjuvant non-covalently incorporated into an intracellular compartment of the cell. 60. The composition of claim 59, wherein said cell is a human cell. 61. The composition of claim 59, wherein said cell is a tumor cell. 62. The composition of claim 61, wherein said cell is a tumor cell listed in Table 2 or Table 3. 63. The composition of claim 61, wherein said cell is an irradiated tumor cell. 64. The composition of claim 61, wherein said cell is a tumor cell that comprises a tumor-associated intracellular antigen. 65. The composition of claim 61, wherein said cell is a tumor cell that comprises a tumor-associated ganglioside antigen. 66. The composition of claim 61, wherein said cell is a melanoma cell. 67. The composition of claim 59, wherein said adjuvant is an adjuvant listed in Table 1. 68. The composition of claim 67, wherein said adjuvant is a detoxified endotoxin. 69. The composition of claim 68, wherein said adjuvant is monophosphoryl lipid A (MPL). 70. A method for stimulating an immune response, comprising administering to an animal an immunologically effective amount of an adjuvant-cell composition comprising a cell that includes an adjuvant non-covalently incorporated into the cell surface membrane of the cell and an adjuvant non-covalently incorporated into an intracellular compartment of the cell. 71. A composition comprising a cell that includes an adjuvant non-covalently incorporated into an intracellular compartment of the cell. 72. The composition of claim 71, wherein said cell is a human cell. 73. The composition of claim 71, wherein said cell is a tumor cell that comprises an intracellular tumor-associated antigen. 74. The composition of claim 73, wherein said cell is a tumor cell that comprises an intracellular tumor-associated antigen listed in Table 12. 75. The composition of claim 73, wherein said cell is an irradiated tumor cell. 76. The composition of claim 71, wherein said adjuvant is monophosphoryl lipid A (MPL). 77. A method for stimulating an immune response, comprising administering to an animal an immunologically effective amount of an adjuvant-cell composition comprising a cell that includes an adjuvant non-covalently incorporated into an intracellular compartment of the cell. 78. A composition comprising an erythrocyte that includes an adjuvant non-covalently incorporated into the erythrocyte cell surface or an intracellular compartment of the erythrocyte. 79. The composition of claim 78, wherein said erythrocyte is a human erythrocyte. 80. The composition of claim 78, wherein said erythrocyte is coated with a tumor-associated antigen. 81. The composition of claim 78, wherein said adjuvant is monophosphoryl lipid A (MPL). 82. A method for stimulating an immune response, comprising administering to an animal an immunologically effective amount of an adjuvant-cell composition comprising an erythrocyte that includes an adjuvant non-covalently incorporated into the erythrocyte cell surface membrane or an intracellular compartment of the erythrocyte. 83. A composition comprising a tumor cell that comprises a tumor-associated ganglioside antigen, the cell including an adjuvant non-covalently incorporated into the cell surface membrane or an intracellular compartment of the cell. 84. The composition of claim 83, wherein said tumor cell is an irradiated tumor cell. 85. A m method for stimulating an immune response, comprising administering to an animal an immunologically effective amount of an adjuvant-cell composition comprising a tumor cell that comprises a tumor-associated ganglioside antigen and that includes an adjuvant non-covalently incorporated into the cell surface membrane or an intracellular compartment of the tumor cell. 86. A composition comprising a melanoma cell that includes an adjuvant non-covalently incorporated into the cell surface or an intracellular compartment of the melanoma cell. 87. The composition of claim 86, wherein said melanoma cell is an irradiated melanoma cell. 88. The composition of claim 86, wherein said melanoma cell is a human melanoma cell. 89. The composition of claim 88, wherein said melanoma cell is the human melanoma cell M27, M18, M14, M111, M22, M7, M102, M108, M16, M104, M109, M25, M24, M10 or M101. 90. The composition of claim 89, wherein said melanoma cell is the human melanoma cell M14, M7, M24, M25, M10 or M101. 91. The composition of claim 86, wherein said adjuvant is monophosphoryl lipid A (MPL). 92. A method for stimulating an immune response, comprising administering to an animal an immunologically effective amount of an adjuvant-cell composition comprising a melanoma cell that includes an adjuvant non-covalently incorporated into the cell surface or an intracellular compartment of the melanoma cell. 93. A composition comprising a cell that includes a detoxified endotoxin adjuvant non-covalently incorporated into the cell surface or an intracellular compartment of the cell. 94. The composition of claim 93, wherein said detoxified endotoxin adjuvant is monophosphoryl lipid A (MPL). 95. The composition of claim 94, prepared by a method comprising the steps of: (a) preparing an MPL-suspended culture media composition by sonicating MPL with a culture medium; and (b) admixing said MPL-suspended culture media composition with a cell composition under conditions effective and for a period of time suitable to allow incorporation of the MPL into the membrane or an intracellular compartment of a cell, thereby preparing said composition. 96. The composition of claim 93, wherein said cell is a human cell. 97. The composition of claim 93, wherein said cell is a tumor cell. 98. The composition of claim 97, wherein said cell is an irradiated tumor cell. 99. The composition of claim 97, wherein said cell is a tumor cell that comprises a tumor-associated intracellular antigen. 100. The composition of claim 97, wherein said cell is a melanoma cell. 101. A method for stimulating an immune response, comprising administering to an animal an immunologically effective amount of an adjuvant-cell composition comprising a cell that includes a detoxified endotoxin adjuvant non-covalently incorporated into the cell surface membrane or an intracellular compartment of the cell. 102. A composition comprising a cell that includes an adjuvant non-covalently incorporated into the cell surface or an intracellular compartment of the cell, the composition prepared by admixing an adjuvant composition with a cell composition under conditions effective and for a period of time suitable to allow incorporation of the adjuvant into the cell surface membrane or an intracellular compartment of the cell. 103. A method for stimulating an immune response, comprising administering to an animal an immunologically effective amount of an adjuvant-cell composition comprising a cell that includes an adjuvant non-covalently incorporated into the cell surface membrane or an intracellular compartment of the cell, the adjuvant-cell composition prepared by admixing an adjuvant composition with a cell composition under conditions effective and for a period of time suitable to allow incorporation of the adjuvant into the cell surface membrane or an intracellular compartment of the cell.

Details for Patent 6,218,166

Applicant	Tradename	Biologic Ingredient	Dosage Form	BLA	Approval Date	Patent No.	Expiredate
Merck Teknika Llc	TICE BCG	bcg live	For Injection	102821	06/21/1989		2018-04-17