Methods and products for inducing mucosal immunity
View Patent ↗The invention relates methods and products for inducing mucosal immunity. In particular, the invention relates to the use of immunostimulatory oligonucleotides containing a CpG motif for inducing mucosal immunity. The CpG immunostimulatory oligonucleotides may be administered alone or in combination with antigen and/or with other adjuvants.
1. A method for inducing a mucosal immune response, comprising:
administering to a subject in need of a mucosal immune response an effective amount for inducing a mucosal immune response of an oligonucleotide 8 to 100 nucleotides in length, having a sequence including at least the following formula:
5′X1X2CGX3X4 3′
wherein C is unmethylated, wherein X1, X2, X3, and X4 are nucleotides, and
an antigen,
wherein the antigen is not encoded in a nucleic acid vector, the oligonucleotide and the antigen are both administered vaginally, rectally, intranasally, ocularly, or by inhalation to the subject, a cytokine and an immune stimulating complex are not administered to the subject, and the antigen is not a Streptococcus pneumoniae antigen.
2. The method of claim 1 , wherein the antigen is administered concurrently with the oligonucleotide.
3. The method of claim 1 , wherein the antigen is delivered in conjunction with a colloidal dispersion system.
4. The method of claim 3 , wherein the colloidal dispersion system is selected from the group consisting of macromolecular complexes, nanocapsules, microspheres, beads, and lipid-based systems.
5. The method of claim 4 , wherein the lipid-based system is selected from the group consisting of oil-in-water emulsions, micelles, mixed micelles, and liposomes.
6. The method of claim 1 , further comprising the step of administering a non-oligonucleotide mucosal adjuvant in conjunction with the antigen.
7. The method of claim 6 , wherein the non-oligonucleotide mucosal adjuvant is selected from the group consisting of cholera toxin, derivatives of cholera toxin, heat-labile enterotoxin, derivatives of heat-labile enterotoxin, alum, monophosphoryl lipid A (MLP), muramyl dipeptide (MDP), saponins, QS21, cytokines, oil-in-water and other emulsion formulations, squalene-in-water emulsion stabilized with Span 85 and Tween 80 (MF59), syntext adjuvant formulation (SAF), Montanide ISA 720 and oil-in-water emulsion containing stabilizing detergent and micelle-forming agent and poly (PCPP) polymers.
8. The method of claim 1 , wherein the subject is a subject at risk of developing an infectious disease.
9. The method of claim 1 , wherein the subject is at risk of developing cancer.
10. The method of claim 1 , wherein the oligonucleotide includes a phosphate backbone modification which is a phosphorothioate or phosphorodithioate modification.
11. The method of claim 10 , wherein the phosphate backbone modification occurs at the 5′ end of the oligonucleotide.
12. The method of claim 10 , wherein the phosphate backbone modification occurs at the 3′ end of the oligonucleotide.
13. The method of claim 1 , wherein X1X2 are nucleotides selected from the group consisting of: GpT, GpG, GpA, ApA, ApT, ApG, CpT, CpA, CpG, TpA, TpT, and TpG; and X3X4 are nucleotides selected from the group consisting of: TpT, CpT, ApT, TpG, ApG, CpG, TpC, ApC, CpC, TpA, ApA, and CpA.
14. The method of claim 1 , wherein the oligonucleotide has a sequence including at least the following formula:
5′TCNTX1X2CGX3X4 3′
wherein X1, X2, X3, and X4 are nucleotides, N is a nucleic acid sequence composed of from about 0-25 nucleotides.
15. The method of claim 1 , wherein the antigen is selected from the group consisting of cells, cell extracts, proteins, polypeptides, peptides, polysaccharides, polysaccharide conjugates, peptide mimics of polysaccharides, lipids, glycolipids, carbohydrates, viruses and viral extracts and parasites.
16. The method of claim 1 , wherein the antigen is obtained from an infectious organism selected from the group consisting of infectious bacteria, infectious viruses, infectious parasites, and infectious fungi.
17. The method of claim 1 , further comprising administering a B-7 costimulatory molecule.
18. The method of claim 1 , wherein the mucosal immune response is induced in a remote site.
19. The method of claim 1 , further comprising administering a boost of the oligonucleotide.
20. The method of claim 6 , further comprising administering a boost of the oligonucleotide and the non-oligonucleotide mucosal adjuvant.
21. The method of claim 1 , further comprising identifying a subject in need of a mucosal immune response.
22. The method of claim 1 , wherein the antigen is a viral antigen.
23. A method for inducing a mucosal immune response, comprising:
administering to a subject in need of a mucosal immune response an effective amount for inducing a mucosal immune response of an oligonucleotide 8 to 100 nucleotides in length, having a sequence including at least the following formula:
5′X1X2CGX3X4 3′
wherein C is unmethylated, wherein X1, X2, X3, and X4 are nucleotides,
a non-oligonucleotide mucosal adjuvant that is not an immune stimulating complex, and
an antigen,
wherein the antigen is not encoded in a nucleic acid vector, and wherein the oligonucleotide, the antigen, and the non-oligonucleotide mucosal adjuvant are all administered rectally, intravaginally, or ocularly, to the subject, and a cytokine is not administered to the subject.
24. A method for inducing a mucosal immune response, comprising:
administering to a subject in need of a mucosal immune response an effective amount for inducing a mucosal immune response of an oligonucleotide 8 to 100 nucleotides in length, having a sequence including at least the following formula:
5′X1X2CGX3X4 3′
wherein C is unmethylated, wherein X1, X2, X3, and X4 are nucleotides, and
a viral antigen,
wherein the antigen is not encoded in a nucleic acid vector, the oligonucleotide and the antigen are both administered vaginally, rectally, intranasally, ocularly, or by inhalation to the subject, and a cytokine and an immune stimulating complex are not administered to the subject.
25. A method for inducing a mucosal immune response, comprising:
administering to a subject in need of a mucosal immune response an effective amount for inducing a mucosal immune response of an oligonucleotide 8 to 100 nucleotides in length, having a sequence including at least the following formula:
5′X1X2CGX3X4 3′
wherein C is unmethylated, wherein X1, X2, X3, and X4 are nucleotides, and
passively exposing the subject to an antigen,
wherein the antigen is not encoded in a nucleic acid vector, oligonucleotide administration and antigen exposure both occur vaginally, rectally, intranasally, or by inhalation, and a cytokine and an immune stimulating complex are not administered to the subject.
26. A method for inducing a mucosal immune response, comprising:
administering to a subject in need of a mucosal immune response an effective amount for inducing a mucosal immune response of an oligonucleotide 8 to 100 nucleotides in length, having a sequence including at least the following formula:
5′X1X2CGX3X4 3′
wherein C is unmethylated, wherein X1, X2, X3, and X4 are nucleotides, and
an antigen,
wherein the antigen is not encoded in a nucleic acid vector, the oligonucleotide and the antigen are both administered vaginally, rectally, or ocularly to the subject, and a cytokine and an immune stimulating complex are not administered to the subject.
27. The method of claim 26 , wherein the antigen is a viral antigen.
28. A method for inducing a mucosal immune response, comprising:
administering to a subject in need of a mucosal immune response an effective amount for inducing a mucosal immune response of an oligonucleotide 8 to 100 nucleotides in length, having a sequence including at least the following formula:
5′X1X2CGX3X4 3′
wherein C is unmethylated, wherein X1, X2, X3, and X4 are nucleotides, and
an antigen,
wherein the antigen is not encoded in a nucleic acid vector and is not a Streptococcus pneumoniae antigen, the oligonucleotide and the antigen are both administered intranasally or by inhalation to the subject, and a cytokine and an immune stimulating complex are not administered to the subject.
29. The method of claim 23 , wherein the antigen is selected from the group consisting of cells, cell extracts, proteins, polypeptides, peptides, polysaccharides, polysaccharide conjugates, peptide mimics of polysaccharides, lipids, glycolipids, carbohydrates, viruses and viral extracts and parasites.
30. The method of claim 25 , wherein the antigen is selected from the group consisting of cells, cell extracts, proteins, polypeptides, peptides, polysaccharides, polysaccharide conjugates, peptide mimics of polysaccharides, lipids, glycolipids, carbohydrates, viruses and viral extracts and parasites.
31. The method of claim 26 , wherein the antigen is selected from the group consisting of cells, cell extracts, proteins, polypeptides, peptides, polysaccharides, polysaccharide conjugates, peptide mimics of polysaccharides, lipids, glycolipids, carbohydrates, viruses and viral extracts and parasites.
32. The method of claim 28 , wherein the antigen is selected from the group consisting of cells, cell extracts, proteins, polypeptides, peptides, polysaccharides, polysaccharide conjugates, peptide mimics of polysaccharides, lipids, glycolipids, carbohydrates, viruses and viral extracts and parasites.
33. A method for inducing a mucosal immune response, comprising
mucosally administering to a subject an oligonucleotide 8 to 100 nucleotides in length and comprising 5′X1X2CGX3X4 3′ wherein C is unmethylated and X1, X2, X3, and X4 are nucleotides, and an antigen that is not encoded in a nucleic acid vector, and
systemically administering to the subject the antigen.
34. The method of claim 33 , wherein the oligonucleotide and antigen are administered mucosally as a priming dose, and the antigen is administered systemically as a boost dose.
35. The method of claim 33 , wherein the antigen is administered systemically as a priming dose, and the oligonucleotide and antigen are administered mucosally as a boost dose.
36. The method of claim 33 , wherein the antigen is administered systemically with a CpG oligonucleotide 8-100 nucleotides in length and comprising 5′X1X2CGX3X4 3′, wherein C is unmethylated, wherein X1, X2, X3, and X4 are nucleotides.
37. The method of claim 33 , wherein the oligonucleotide and antigen are administered mucosally with a non-nucleic acid mucosal adjuvant.
38. The method of claim 33 , wherein the antigen is administered systemically with a non-nucleic acid mucosal adjuvant.
39. The method of claim 26 , wherein the antigen is administered systemically with a non-nucleic acid mucosal adjuvant.
40. The method of claim 33 , wherein the antigen is a polypeptide or a peptide.
41. The method of claim 33 , wherein mucosally administering is intranasally administering or administering by inhalation.
42. The method of claim 40 , wherein mucosally administering is intranasally administering or administering by inhalation.
43. The method of claim 33 , wherein systemically administering is intramuscularly administering.
44. The method of claim 40 , wherein systemically administering is intramuscularly administering.
45. The method of claim 42 , wherein systemically administering is intramuscularly administering.
46. A method for inducing an immune response, comprising
systemically administering to a subject, as a priming dose, an antigen that is not encoded in a nucleic acid vector, and
mucosally administering to the subject, as a boost dose, an oligonucleotide 8-100 nucleotides in length and comprising 5′X1X2CGX3X4 3′ wherein C is unmethylated and X1, X2, X3, and X4 are nucleotides, and the antigen.
47. The method of claim 46 , wherein the antigen is systemically administered with a CpG oligonucleotide 8-100 nucleotides in length and comprising 5′X1X2CGX3X4 3′ wherein C is unmethylated and X1, X2, X3, and X4 are nucleotides.
48. The method of claim 46 , wherein the antigen is systemically administered with a non-nucleic acid mucosal adjuvant.
49. The method of claim 47 , wherein the antigen is systemically administered with a non-nucleic acid mucosal adjuvant.
50. The method of claim 46 , wherein the antigen is mucosally administered to the subject with a non-nucleic acid mucosal adjuvant.
51. The method of claim 49 , wherein the antigen is mucosally administered to the subject with a non-nucleic acid mucosal adjuvant.
52. A method for inducing a mucosal immune response in a subject, comprising
mucosally administering to a subject, as a boost dose, an oligonucleotide 8-100 nucleotides in length and comprising 5′X1X2CGX3X4 3′ wherein C is unmethylated and X1, X2, X3, and X4 are nucleotides, and an antigen that is not encoded by a nucleic acid vector, wherein the subject has received a priming dose of antigen administered systemically.