Biodegradable immunomodulatory formulations and methods for use thereof
The invention provides new compositions and methods for immunomodulation of individuals. Immunomodulation is accomplished by administration of immunomodulatory polynucleotide/microcarrier (IMP/MC) complexes. The IMP/MC complexes may be covalently or non-covalently bound, and feature a polynucleotide comprising at least one immunostimulatory sequence bound to a biodegradable microcarrier or noncarrier.
1. An immunomodulatory polynucleotide/microcarrier (IMP/MC) complex, comprising:
a polynucleotide covalently linked to the surface of a biodegradable microcarrier (MC), wherein said polynucleotide comprises the sequence 5′-CG-3′, wherein the polynucleotide is greater than 6 and less than about 200 nucleotides in length and wherein said MC is less than 10 μm in size.
2. The IMP/MC complex of claim 1 , wherein said microcarrier is a liquid phase microcarrier.
3. The IMP/MC complex of claim 1 , wherein said microcarrier is a solid phase microcarrier.
4. The IMP/MC complex of claim 1 , wherein said microcarrier is from 25 nm to 5 μm in size.
5. The IMP/MC complex of claim 4 , wherein said microcarrier is from 1.0 μm to 2.0 μm in size.
6. The IMP/MC complex of claim 5 , wherein said microcarrier is 1.4 μm in size.
7. The IMP/MC complex of claim 1 , wherein said microcarrier is cationic.
8. The IMP/MC complex of claim 1 , wherein said complex is antigen-free.
9. The IMP/MC complex of claim 1 , wherein said polynucleotide comprises the sequence 5′-T, C, G-3′.
10. The IMP/MC complex of claim 9 , wherein said polynucleotide comprises the sequence 5′-TCGX 1 X 2 X 3 X 4 -3′ or the sequence 5′-X 1 TCGX 2 X 3 X 4 -3′, wherein X 1 , X 2 , X 3 , X 4 are nucleotides.
11. The IMP/MC complex of claim 10 , wherein said polynucleotide comprises the sequence 5′-TCGTCGX 4 -3 ′.
12. The IMP/MC complex of claim 10 , wherein said polynucleotide comprises a sequence selected from the group consisting of 5′-TCGTCGA-3′, 5′-TCGAAAA-3′, 5′-TCGCCCC-3′, 5′-TCGGGGG-3′ and 5′-TCGTTTT-3′.
13. The IMP/MC complex of claim 1 , wherein said polynucleotide comprises the sequence 5′-C, G, pyrimidine, pyrimidine, C, G-3′.
14. The IMP/MC complex of claim 1 , wherein said polynucleotide comprises the sequence 5′-purine, purine, C, G, pyrimidine, pyrimidine, C, G-3′.
15. The IMP/MC complex of claim 9 , wherein said polynucleotide comprises the sequence SEQ ID NO:1.
16. The IMP/MC complex of any of claims 1 , 9 , 10 , 11 , or 12 , wherein said polynucleotide is 7 nucleotides in length.
17. The IMP/MC complex of any of claims 1 , 9 , 10 , 11 , or 12 , wherein said complex further comprises an antigen.
18. The IMP/MC complex of claim 17 , wherein said antigen is an allergen.
19. The IMP/MC complex of claim 1 , wherein said polynucleotide comprises a phosphate backbone modification.
20. The IMP/MC complex of claim 19 , wherein said phosphate backbone modification is a phosphorothioate.
21. A kit, comprising:
an immunomodulatory polynucleotide/microcarrier (IMP/MC) complex, said complex comprising a polynucleotide covalently linked the surface of to a biodegradable microcarrier (MC), wherein said polynucleotide comprises the sequence 5′-CG-3′, wherein the polynucleotide is greater than 6 and less than about 200 nucleotides in length and wherein said MC is less than 10 μm in size; and instructions for use of the IMP/MC complex in immunomodulation of an individual.
22. The kit of claim 21 , wherein said microcarrier is a liquid phase microcarrier.
23. The kit of claim 21 , wherein said microcarrier is a solid phase microcarrier.
24. The kit of claim 21 , wherein said microcarrier is from 25 nm to 5 μm in size.
25. The kit of claim 24 , wherein said microcarrier is from 1.0 μm to 2.0 μm in size.
26. The kit of claim 25 , wherein said microcarrier is 1.4 μm in size.
27. The kit of claim 21 , wherein said microcarrier is cationic.
28. The kit of claim 21 , wherein said complex is antigen-free.
29. The kit of claim 21 , wherein said polynucleotide comprises the sequence 5′-T, C, G-3′.
30. The kit of claim 29 , wherein said polynucleotide comprises the sequence 5′-TCGX1X2X3X4-3′ or the sequence 5′X1TCGX2X3X4-3′, wherein X1, X2, X3, X4 are nucleotides.
31. The kit of claim 29 , wherein said polynucleotide comprises the sequence 5′-TCGTCGX 4 -3′.
32. The kit of claim 30 , wherein said polynucleotide comprises a sequence selected from the group consisting of 5′-TCGTCGA-3′, 5′-TCGAAAA-3′, 5′-TCGCCCC-3′, 5′-TCGGGGG-3′ and 5′-TCGTTTT-3′.
33. The kit of claim 21 , wherein the polynucleotide comprises the sequence 5′-C, G, pyrimidine, pyrimidine, C, G-3′.
34. The kit of claim 21 , wherein the polynucleotide comprises the sequence 5′-purine, purine, C, G, pyrimidine, pyrimidine, C, G-3′.
35. The kit of claim 29 , wherein the polynucleotide comprises the sequence SEQ ID NO:1.
36. The kit of any of claims 21 , 29 , 30 , 31 , or 32 , wherein said kit further comprises an antigen.
37. The kit of claim 36 , wherein said antigen is an allergen.
38. The kit of claim 21 , wherein said polynucleotide comprises a phosphate backbone modification.
39. The kit of claim 38 , wherein said phosphate backbone modification is a phosphorothioate.
40. A kit, comprising:
an immunomodulatory polynucleotide/microcarrier (IMP/MC) complex, said complex comprising a polynucleotide covalently linked to the surface of a biodegradable microcarrier (MC), wherein said polynucleotide comprises the sequence 5′-CG-3′ and wherein said polynucleotide is 7 nucleotides in length; and instructions for use of the IMP/MC complex in immunomodulation of an individual.
41. The kit of claim 40 , wherein said polynucleotide comprises the sequence 5′-T, C, G-3′.
42. The kit of claim 41 , wherein said polynucleotide consists of the sequence 5′-TCGX 1 X 2 X 3 X 4 -3′ or the sequence 5′-X 1 TCGX 2 X 3 X 4 -3′, wherein X 1 , X 2 , X 3 , X 4 are nucleotides.
43. The kit of claim 42 , wherein said polynucleotide consists of the sequence 5′-TCGTCGX 4 -3 ′.
44. The kit of claim 42 , wherein said polynucleotide consists of a sequence selected from the group consisting of 5′-TCGTCGA-3′, 5′-TCGAAAA-3′, 5′-TCGCCCC-3′, 5′-TCGGGGG-3′ and 5′-TCGTTTT-3′.
45. The kit of claim 40 , wherein said complex is antigen-free.
46. The kit of claim 40 , further comprising an antigen.
47. The kit of claim 46 , wherein said antigen is an allergen.
48. The kit of claim 40 , wherein said polynucleotide comprises a phosphate backbone modification.
49. The kit of claim 48 , wherein said phosphate backbone modification is a phosphorothioate.
50. A composition comprising an IMP/MC complex of claim 1 and a pharmaceutically acceptable excipient.
51. A composition according to claim 50 , wherein the composition is antigen-free.
52. A composition according to claim 50 , wherein the composition further comprises an antigen.
53. A composition according to claim 52 , wherein the antigen is an allergen.