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 method of modulating an immune response in an individual comprising administering to an individual a composition comprising an immunomodulatory polynucleotide/microcarrier (IMP/MC) complex, said complex comprising a polynucleotide linked to 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, in an amount sufficient to modulate an immune response in said individual.
2. The method of claim 1 , wherein said microcarrier is a solid phase microcarrier.
3. The method of claim 1 , wherein said microcarrier is a liquid phase microcarrier.
4. The method of claim 1 , wherein said polynucleotide is covalently linked to said microcarrier.
5. The method of claim 1 , wherein said polynucleotide is non-covalently linked to said microcarrier.
6. The method of claim 1 , wherein said complex is antigen-free.
7. The method of claim 1 , wherein a Th1-type immune response is stimulated.
8. The method of claim 1 , wherein a Th2-type immune response is suppressed.
9. The method of claim 1 , wherein interferon-gamma (IFN-γ) is increased in said individual.
10. The method of claim 1 , wherein interferon-alpha (IFN-α) is increased in said individual.
11. The method of claim 1 , wherein levels of IgE is reduced in said individual.
12. The method of claim 1 , wherein said polynucleotide comprises the immunostimulatory sequence 5′-T, C, G-3′.
13. The method of claim 12 , 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.
14. The method of claim 13 , wherein the polynucleotide comprises the immunostimulatory sequence 5′-TCGTCGX 1 -3′, wherein X 1 is a nucleotide.
15. The method of claim 13 , 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′.
16. The method of claim 1 , wherein said polynucleotide comprises the immunostimulatory sequence 5′-C, G, pyrimidine, pyrimidine, C, G-3′.
17. The method of claim 1 , wherein said polynucleotide comprises the immunostimulatory sequence 5′-purine, purine, C, G, pyrimidine, pyrimidine, C, G-3′.
18. The method of claim 12 , wherein said polynucleotide comprises the immunostimulatory sequence SEQ ID NO:1.
19. The method of any of claims 1 , or 12 - 15 wherein said polynucleotide is 7 nucleotides in length.
20. The method of any of claims 1 , or 12 - 15 wherein said composition further comprises an antigen.
21. The method of claim 20 , wherein said antigen is an allergen.
22. The method of claim 1 , wherein said polynucleotide comprises a phosphate backbone modification.
23. The method of claim 22 , wherein said phosphate backbone modification is a phosphorothioate.