MODULATION OF NKT CELL ACTIVITY WITH ANTIGEN-LOADED CD1d MOLECULES

Abstract

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 - 77 . (canceled) 
     
     
         78 . A* composition comprising:
 (a) an immunogen;   (b) a CD1d complex, said complex comprising:
 (i) an isolated soluble CD1d polypeptide sufficient to associate with β2-microglobulin and bind a ceramide-like glycolipid antigen; 
 (ii) an isolated polypeptide comprising β2-microglobulin or a fragment thereof associated with said CD1d polypeptide; and 
 (iii) a ceramide-like glycolipid antigen bound to said CD1d polypeptide; and 
   (c) a carrier.   
     
     
         79 . The composition of  claim 78 , wherein the composition additionally comprises an adjuvant. 
     
     
         80 . The composition of  claim 79 , wherein the adjuvant is a TLR agonist selected from the group consisting of: MPL, CpG, and BCG. 
     
     
         81 . The composition of  claim 80 , wherein the adjuvant is MPL. 
     
     
         82 . The composition of  claim 78 , wherein the ceramide-like glycolipid antigen comprises an α-galactosylceramide. 
     
     
         83 . The composition of  claim 78 , wherein the soluble CD1d polypeptide comprises an amino acid sequence at least 90% identical to amino acids 21 to 295 of SEQ ID NO: 1. 
     
     
         84 . The composition of  claim 78 , wherein the soluble CD1d polypeptide comprises an amino acid sequence at least 95% identical to amino acids 21 to 295 of SEQ ID NO: 1. 
     
     
         85 . The composition of  claim 78 , wherein the soluble CD1d polypeptide comprises amino acids 21 to 295 of SEQ ID NO: 1. 
     
     
         86 . The composition of  claim 78 , wherein the soluble CD1d polypeptide comprises amino acids 1 to 295 of SEQ ID NO: 1. 
     
     
         87 . The composition of  claim 78 , wherein the 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. 
     
     
         88 . The composition of  claim 78 , wherein the soluble CD1d polypeptide further comprises a heterologous polypeptide. 
     
     
         89 . The composition of  claim 78 , wherein the β2-microglobulin polypeptide comprises an amino acid sequence at least 90% identical to amino acids 21 to 113 of SEQ ID NO: 2. 
     
     
         90 . The composition of  claim 89 , wherein the β2-microglobulin polypeptide comprises amino acids 21 to 113 of SEQ ID NO: 2. 
     
     
         91 . The composition of  claim 89 , wherein the β2-microglobulin polypeptide comprises amino acids 1 to 119 of SEQ ID NO: 2. 
     
     
         92 . The composition of  claim 82 , wherein the 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. 
       
     
     
         93 . The composition of  claim 78 , wherein the composition further comprises a costimulatory molecule. 
     
     
         94 . The composition of  claim 93 , wherein the costimulatory molecule is a Saponin, a toll-like receptor (“TLR”), B7-1 (CD80) or B7-2 (CD86).