Immune potentiating compositions of cancer cells
A composition of matter is disclosed, comprising an immunostimulatory molecule and animal cells cultured in the presence of at least one interferon (IFN) for a time and under conditions sufficient to enhance the antigen presenting function of said cells. Also disclosed are immunopotentiating compositions and their use for treatment and/or prophylaxis of a disease or condition.
1. A method for enhancing immunopotentiation of animal cells, comprising:
culturing animal cells in the presence of a T-cell co-stimulatory B7molecule selected from the group consisting of B7-1 and B7-2 on the surface of said cells in the presence of a type II interferon (IFN) in exogenous form and at least one type I IFN in exogenous form for a time and under conditions sufficient to enhance the antigen presenting functions of said cells.
2. The method of claim 1 , wherein the animal cells are cultured by contacting said cells with a type II IFN in exogenous form for a time and under conditions sufficient to permit cellular responsiveness to at least one type I IFN and contacting the cells with the at least one type I IFN in exogenous form for a time and under conditions sufficient to enhance the antigen presenting function of said cells.
3. The method of claim 1 , wherein the type II IFN is present in the culture medium at a concentration of about 100 to about 2000 international units/mL.
4. The method of claim 1 , wherein an individual type I IFN is present in the culture medium at a concentration of about 100 to about 2000 international units/mL.
5. The method of claim 1 , wherein the cells are cultured in the presence of a type II IFN from about 16 to about 96 hours and subsequently in the presence of at least one type I IFN from about 16 to about 72 hours.
6. The method of claim 1 , wherein said animal cells express said T-cell co-stimulatory B7 molecule.
7. The method of claim 6 , further comprising modifying the animal cells to express said T-cell co-stimulatory B7 molecule prior to culturing.
8. The method of claim 7 , wherein said modification comprises introducing into said animal cells a polynucleotide from which the T-cell co-stimulatory B7molecule is expressed.
9. The method of claim 1 , wherein the type II IFN is an IFN-gamma.
10. The method of claim 1 , wherein an individual type I IFN is selected from the group consisting of an IFN-alpha and an IFN-beta
11. The method of claim 1 , further comprising inactivating said cells.
12. The method of claim 11 , wherein said inactivation comprises treating said cells to render them incapable of proliferation.
13. The method of claim 9 , wherein the IFN-gamma comprises the amino acid sequence set forth in SEQ ID NO: 2.
14. The method of claim 10 , wherein the IFN-beta is an IFN-beta 1.
15. The method of claim 10 , wherein the IFN-beta is an IFN-beta 1comprising the amino acid sequence set forth in SEQ ID NO: 4.
16. The method of claim 10 , wherein the IFN-beta is an IFN-beta 2.
17. The method of claim 10 , wherein the IFN-beta is an IFN-beta 2comprising the amino acid sequence set forth in SEQ ID NO: 6.
18. The method of claim 10 , wherein the IFN-alpha comprises the amino acid sequence set forth in SEQ ID NO: 8.
19. The method of claim 10 , wherein the IFN-alpha is an IFN-alpha 1.
20. The method of claim 10 , wherein the IFN-alpha 1 comprises the amino acid sequence set forth in SEQ ID NO: 10.
21. The method of claim 10 , wherein the IFN-alpha is an IFN-alpha 2.
22. The method of claim 10 , wherein the IFN-alpha is an IFN-alpha 2comprising the amino acid sequence set forth in SEQ ID NO: 12.
23. The method of claim 1 , wherein the animal cells are cultured by contacting the cells with a single type I IFN in exogenous form.
24. The method of claim 23 , wherein said type I IFN is selected from the group consisting of an IFN-alpha and an IFN-beta.
25. The method of claim 24 , wherein said type I IFN is an IFN-beta.
26. The method of claim 1 , wherein the animal cells are cultured by contacting the cells with a first type I IFN in exogenous form and a second type I IFN in exogenous form.
27. The method of claim 26 , wherein the first type I IFN and the second type I IFN are each independently selected from the group consisting of an IFN-beta and an IFN-alpha.
28. The method of claim 26 , wherein the type II IFN is an IFN-gamma and wherein the first type I IFN and the second type I IFN are individually selected from IFN-alpha and IFN-beta.
29. The method of claim 26 , wherein the type II IFN is an IFN-gamma and wherein the first type I IFN is an IFN-beta and the second type I IFN is an IFN-alpha.
30. The method of claim 1 , wherein the cells are cultured in the presence of IFN-gamma from about 48 to about 96 hours and subsequently in the presence of IFN-alpha and/or IFN-beta from about 24 to about 72 hours.
31. The method of claim 1 , wherein the cells are cultured in the presence of IFN-gamma from about 48 to about 96 hours and subsequently in the presence of IFN-beta from about 24 to about 72 hours.
32. The method for enhancing immunopotentiation of animal cells of claim 1 , further comprising:
combining the animal cells with the a T-cell co-stimulatory B7 molecule in soluble form prior to culturing.
33. The method of claim 32 , wherein the animal cells are cultured by contacting said cells with a type II IFN in exogenous form for a time and under conditions sufficient to permit cellular responsiveness to at least one type I IFN and contacting the cells with the at least one type I IFN in exogenous form for a time and under conditions sufficient to enhance the antigen presenting function of said cells.
34. The method of claim 32 , wherein the type II IFN is present in the culture medium at a concentration of about 100 to about 2000 international units/mL.
35. The method of claim 32 , wherein an individual type I IFN is present in the culture medium at a concentration of about 100 to about 2000 international units/mL.
36. The method of claim 32 , wherein the cells are cultured in the presence of a type II IFN from about 16 to about 96 hours and subsequently in the presence of at least one type I IFN from about 16 to about 72 hours.
37. The method of claim 32 , wherein the type II IFN is an IFN-gamma.
38. The method of claim 32 , wherein an individual type I IFN is selected from the group consisting of an IFN-alpha and an IFN-beta.
39. The method of claim 32 , further comprising inactivating said cells.
40. The method of claim 39 , wherein said inactivation comprises treating said cells to render them incapable of proliferation.
41. The method of claim 37 , wherein the IFN-gamma comprises the amino acid sequence set forth in SEQ ID NO: 2.
42. The method of claim 38 , wherein the IFN-beta is an IFN-beta 1.
43. The method of claim 38 , wherein the IFN-beta is an IFN-beta 1comprising the amino acid sequence set forth in SEQ ID NO: 4.
44. The method of claim 38 , wherein the IFN-beta is an IFN-beta 2.
45. The method of claim 38 , wherein the IFN-beta is an IFN-beta 2comprising the amino acid sequence set forth in SEQ ID NO: 6.
46. The method of claim 38 , wherein the IFN-alpha comprises the amino acid sequence set forth in SEQ ID NO: 8.
47. The method of claim 38 , wherein the IFN-alpha is an IFN-alpha 1.
48. The method of claim 38 , wherein the IFN-alpha is an IFN-alpha 1comprising the amino acid sequence set forth in SEQ ID NO: 10.
49. The method of claim 38 , wherein the IFN-alpha is an IFN-alpha 2.
50. The method of claim 38 , wherein the IFN-alpha is an IFN-alpha 2comprising the amino acid sequence set forth in SEQ ID NO: 12.
51. The method of claim 32 , wherein the animal cells are cultured by contacting the cells with a single type I IFN in exogenous form.
52. The method of claim 51 , wherein said type I IFN is selected from the group consisting of an IFN-alpha and an IFN-beta.
53. The method of claim 52 , wherein said type I IFN is an IFN-beta.
54. The method of claim 32 , wherein the animal cells are cultured by contacting the cells with a first type I IFN in exogenous form and a second type I IFN in exogenous form.
55. The method of claim 54 , wherein the first type I IFN and the second type I IFN are each independently selected from the group consisting of an IFN-beta and an IFN-alpha.
56. The method of claim 54 , wherein the type II IFN is an IFN-gamma and wherein the first type I IFN and the second type I IFN are individually selected from IFN-alpha and IFN-beta.
57. The method of claim 54 , wherein the type II IFN is an IFN-gamma and wherein the first type I IFN is an IFN-beta and the second type I IFN is an IFN-alpha.
58. The method of claim 32 , wherein the cells are cultured in the presence of IFN-gamma from about 48 to about 96 hours and subsequently in the presence of IFN-alpha and/or IFN-beta from about 24 to about 72 hours.