Modulation of NKT cell activity with antigen-loaded CD1d molecules
View Patent ↗The invention is directed to methods of modulating an immune response in an animal, comprising administering a composition comprising one or more soluble CD1d complexes, in particular non-specific soluble CD1d complexes. Soluble CD1d complexes comprise a soluble CD1d polypeptide, a β2-microglobulin polypeptide, and a ceramide-like glycolipid antigen bound to the CD1d antigen binding groove, and in certain embodiments, an immunogen. The administration of compositions of the present invention affects the activity of CD1d-restricted NKT cells, and in particular, allows for multiple administrations without causing CD1d-restricted NKT cell anergy.
1. A method of enhancing an immune response to a polypeptide immunogen in an animal, comprising administering to an animal in need thereof a composition comprising:
(a) a polypeptide immunogen;
(b) a nonspecific CD1d complex, said complex comprising:
(i) a soluble CD1d polypeptide sufficient to associate with β2-microglobulin and bind a ceramide-like glycolipid antigen;
(ii) a polypeptide comprising β2-microglobulin or a fragment thereof associated with said CD1d polypeptide;
(iii) a ceramide-like glycolipid antigen bound to said CD1d polypeptide, wherein said ceramide-like glycolipid antigen has an α-linked galactose or glucose and is capable of activating NKT cells; and
(c) a carrier;
wherein said CD1d complex is administered in an amount sufficient to enhance the immune response against said polypeptide immunogen relative to administration of said polypeptide immunogen in the absence of said CD1d complex.
2. The method of claim 1 , wherein said composition additionally comprises an adjuvant.
3. The method of claim 1 , wherein said ceramide-like glycolipid antigen comprises an α-galactosylceramide.
4. The method of claim 2 , wherein said adjuvant is a Toll-like receptor (TLR) agonist selected from the group consisting of: monophosphoryl lipid A (MPL), CpG, and BCG.
5. The method of claim 4 , wherein said adjuvant is MPL.
6. The method of claim 1 , wherein said soluble CD1d polypeptide comprises an amino acid sequence at least 90% identical to amino acids 21 to 295 of SEQ ID NO: 1.
7. The method of claim 6 , wherein said soluble CD1d polypeptide comprises an amino acid sequence at least 95% identical to amino acids 21 to 295 of SEQ ID NO: 1.
8. The method of claim 7 , wherein said soluble CD1d polypeptide comprises amino acids 21 to 295 of SEQ ID NO: 1.
9. The method of claim 7 , wherein said soluble CD1d polypeptide comprises amino acids 1 to 295 of SEQ ID NO: 1.
10. The method of claim 1 , wherein said soluble CD1d polypeptide comprises amino acids 20 to 295 of SEQ ID NO: 1, except for at least one but less than 10 conservative amino acid substitutions.
11. The method of claim 1 , wherein said soluble CD1d polypeptide further comprises a heterologous polypeptide.
12. The method of claim 1 , wherein said β2-microglobulin polypeptide comprises an amino acid sequence at least 90% identical to amino acids 21 to 113 of SEQ ID NO: 2.
13. The method of claim 1 , wherein said β2-microglobulin polypeptide comprises amino acids 21 to 113 of SEQ ID NO: 2.
14. The method of claim 1 , wherein said β2-microglobulin polypeptide comprises amino acids 1 to 119 of SEQ ID NO: 2.
15. The method of claim 3 , wherein said α-galactosylceramide comprises the formula:
wherein
R1 is a linear or branched C 2 -C 27 alkene with at least one C═C bond but not —(CH 2 ) 7 CH═CH(CH 2 ) 7 CH 3 , or R1 is C(OH) R3 wherein R3 is a linear or branched C 2 -C 26 alkene with at least one C═C bond; and
R2 is one of the following (a)-(e):
(a) —CH 2 (CH 2 ) x ,CH 3 ,
(b) —CH(OH)(CH 2 ) x CH 3 ,
(c) —CH(OH)(CH 2 ) x (CH)(CH 3 ) 2 ,
(d) —CH═CH(CH 2 ) x CH 3 , and
(e) —CH(OH)(CH 2 ) x CH(CH 3 )CH 2 CH 3 ,
wherein X is an integer ranging from 5-17.
16. The method of claim 1 , wherein said composition further comprises a costimulatory molecule.
17. The method of claim 16 , wherein said costimulatory molecule is B71.
18. A method of enhancing an immune response to a polypeptide immunogen in an animal and inhibiting an anergic effect of a ceramide-like glycolipid antigen on NKT cell activity, said method comprising:
(a) administering to an animal in need thereof a composition comprising:
(i) a polypeptide immunogen;
(ii) a nonspecific CD1d complex, said complex comprising:
(1) a soluble CD1d polypeptide sufficient to associate with β2microglobulin and bind a ceramide-like glycolipid antigen; and
(2) a polypeptide comprising β2-microglobulin or a fragment thereof associated with said CD1d polypeptide;
(3) a ceramide-like glycolipid antigen bound to said CD1d polypeptide, wherein said ceramide-like glycolipid antigen has an α-linked galactose or glucose and is capable of activating NKT cells; and
(iii) a carrier;
wherein said CD1d complex is administered in an amount sufficient to enhance the immune response against said polypeptide immunogen relative to administration of said polypeptide immunogen in the absence of said CD1d complex; and
(b) readministering said composition to said animal one or more times, wherein said NKT cells are activated in response to readministration of said composition.