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 nanocarrier.
1. A composition comprising an immunomodulatory polynucleotide/microcarrier (IMP/MC) complex, said complex comprising a polynucleotide noncovalently-linked to the surface of a biodegradable microcarrier (MC), wherein said polynucleotide comprises an immunostimulatory sequence (ISS) 5′-C,G-3′, wherein the C is an unmethylated cytosine, wherein the polynucleotide is greater than 6 nucleotides and less than about 200 nucleotides in length, and wherein said MC is less than 10 μm in size.
2. The composition of claim 1 , wherein said microcarrier is a solid phase microcarrier.
3. The composition of claim 1 , wherein said microcarrier is a liquid phase microcarrier.
4. The composition of claim 1 , wherein said polynucleotide comprises a phosphate backbone modification.
5. The composition of claim 4 , wherein said phosphate backbone modification is a phosphorothioate.
6. The composition of claim 1 , wherein said complex is antigen-free.
7. The composition of claim 1 , wherein said microcarrier is cationic.
8. The composition of claim 7 , wherein said microcarrier is selected from the group consisting of poly(lactic acid), poly(glycolic acid), copolymers of poly(lactic acid) and poly(glycolic acid), block copolymers of poly(lactic acid) and polyethylene glycol, polyorthoesters, polyanhydrides, polyanhydride imides, polyanhydride esters, polyphosphazenes, polyamides, liposomes, micelles, and oil droplets.
9. The composition of claim 1 , wherein said polynucleotide comprises the immunostimulatory sequence 5′-T, C, G-3′.
10. The composition of claim 9 , wherein said polynucleotide comprises the immunostimulatory sequence 5′-TCGX 1 X 2 X 3 X 4 -3′ or the immunostimulatory sequence 5′-X 1 TCGX 2 X 3 X 4 -3′, wherein X 1 , X 2 , X 3 , X 4 are nucleotides.
11. The composition of claim 10 , wherein the polynucleotide comprises the immunostimulatory sequence 5′-TCGTCGX 1 -3′, wherein X 1 is a nucleotide.
12. The composition of claim 10 , wherein said polynucleotide comprises an immunostimulatory 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 composition of claim 1 , wherein said polynucleotide comprises the immunostimulatory sequence 5′-C, G, pyrimidine, pyrimidine, C, G-3′.
14. The composition of claim 1 , wherein said polynucleotide comprises the immunostimulatory sequence 5′-purine, purine, C, G, pyrimidine, pyrimidine, C, G-3′.
15. The composition of claim 9 , wherein said polynucleotide comprises the immunostimulatory sequence SEQ ID NO:1.
16. The composition of any of claim 1 , 9 , 10 , 11 or 12 , wherein said polynucleotide is 7 nucleotides in length.
17. The composition of any of claim 1 , 9 , 10 , 11 or 12 , wherein said composition further comprises an antigen.
18. The composition of claim 17 , wherein said antigen is an allergen.
19. The composition of claim 1 , wherein said microcarrier is from 25 nm to 5 μm in size.
20. The composition of claim 19 , wherein said microcarrier is from 1.0 μm to 2.0 μm in size.
21. The composition of claim 1 , wherein said microcarrier is from 400 nm to less than 10 μm in size.
22. The composition of claim 1 , further comprising a pharmaceutically acceptable excipient.