Trimeric OX40-immunoglobulin fusion protein and methods of use

Abstract

Compositions including a trimeric OX-40 fusion protein are disclosed. Also disclosed are methods for enhancing the immune response of a mammal to an antigen by engaging the OX-40 receptor on the surface of T-cells involving administering to the mammal a composition comprising a trimeric OX-40 fusion protein and a pharmaceutically acceptable carrier.

1 . A fusion polypeptide comprising in an N-terminal to C-terminal direction: 
 an immunoglobulin domain;    a trimerization domain; and    a receptor binding domain, wherein the fusion polypeptide self-assembles into a trimeric fusion protein.    
     
     
         2 . The fusion polyeptide of  claim 1 , wherein the receptor binding domain comprises a domain of a Tumor Necrosis Family ligand.  
     
     
         3 . The fusion polypeptide of  claim 2 , wherein the receptor binding domain is an OX-40L receptor binding domain.  
     
     
         4 . The fusion polypeptide of  claim 3 , wherein the fusion protein is capable of binding to the OX-40 receptor and stimulating at least one OX-40 mediated activity.  
     
     
         5 . The fusion polypeptide of  claim 4 , OX-40 mediated activity is CD4 +  T cell proliferation.  
     
     
         6 . The fusion polypeptide of  claim 3 , wherein the OX-40 receptor binding domain comprises an extracellular domain of OX-40 ligand (OX-40L).  
     
     
         7 . The fusion polypeptide of  claim 6 , wherein the OX-40 receptor binding domain comprises a polynucleotide sequence at least 95% identical to SEQ ID NO:2.  
     
     
         8 . The fusion polypeptide of  claim 1 , wherein the trimerization domain comprises an isoleucine zipper domain.  
     
     
         9 . The fusion polypeptide of  claim 1 , wherein the isoleucine zipper domain comprises a polynucleotide sequence at least 95% identical to SEQ ID NO:4.  
     
     
         10 . The fusion polypeptide of  claim 1 , wherein the immunoglobulin domain comprises an Fc domain.  
     
     
         11 . The fusion polypeptide of  claim 1 , comprising an amino acid sequence at least 95% identical to SEQ ID NO:8.  
     
     
         12 . The fusion polypeptide of  claim 11 , wherein the polypeptide is encoded by a nucleic acid that hybridizes under highly stringent conditions to a nucleic acid with the polynucleotide sequence of SEQ ID NO:7.  
     
     
         13 . A fusion protein comprising a plurality of the fusion polypeptides of  claim 1 .  
     
     
         14 . The fusion protein of  claim 13 , consisting of three or six fusion polypeptides.  
     
     
         15 . The fusion protein of  claim 13 , comprising an OX-40L fusion protein.  
     
     
         16 . A recombinant nucleic acid comprising a polynucleotide sequence that encodes the fusion polypeptide of  claim 1 .  
     
     
         17 . The recombinant nucleic acid of  claim 16 , the polynucleotide sequence comprising in a 5′ to 3′ direction: 
 a polynucleotide sequence encoding an immunoglobulin domain;    a polynucleotide sequence encoding a trimerization domain; and    a polynucleotide encoding a ligand receptor binding domain.    
     
     
         18 . The recombinant nucleic acid of  claim 17 , wherein the polynucleotide encoding a ligand receptor binding domain encodes an OX-40L domain.  
     
     
         19 . A pharmaceutical composition comprising the fusion polypeptide of  claim 1 , or a nucleic acid encoding the fusion polypeptide.  
     
     
         20 . A fusion polypeptide comprising in an N-terminal to C-terminal direction: 
 a dimerization domain;    a trimerization domain; and    an OX-40 receptor binding domain, wherein the fusion polypeptide self-assembles into a trimeric fusion protein.    
     
     
         21 . The fusion polypeptide of  claim 20 , wherein the dimerization domain is an immunoglobulin domain.  
     
     
         22 . The fusion polypeptide of  claim 21 , wherein the immunoglobulin domnain is an Fc domain.  
     
     
         23 . The fusion polypeptide of  claim 20 , wherein the trimerization domain is an isoleucine zipper domain.  
     
     
         24 . The fusion polypeptide of  claim 20 , wherein the trimeric fusion protein consists of three or six fusion polypeptides.  
     
     
         25 . A method of enhancing an immune response in a subject, the method comprising: 
 administering to a subject exposed to an antigen, a therapeutically effective amount of the fusion protein of  claim 1 , thereby enhancing the immune response to the antigen by the subject.    
     
     
         26 . The method of  claim 25 , wherein the subject is a human subject.  
     
     
         27 . The method of  claim 25 , wherein the subject is exposed to the antigen prior to administering theusion protein.  
     
     
         28 . The method of  claim 25 , wherein the antigen and the trimeric OX-40L fusion protein are administered at the same time.  
     
     
         29 . The method of  claim 25 , wherein the fusion protein is administered by expressing a recombinant nucleic acid encoding an OX-40L fusion polypeptide, which fusion polypeptide assembles into the fusion protein in at least one cell of the subject.  
     
     
         30 . The method of  claim 29 , wherein the recombinant nucleic acid encoding the OX-40L fusion polypeptide is introduced ex vivo into at least one cell, and the at least one cell comprising the recombinant nucleic acid is introduced into the subject.  
     
     
         31 . The method of  claim 29 , comprising administering the fusion protein by introducing into the subject a viral or plasmid vector comprising the nucleic acid encoding an OX-40L fusion polypeptide, which fusion polypeptide assembles into the fusion protein.  
     
     
         32 . The method of  claim 31 , wherein the viral vector is an adenovirus vector, a retrovirus vector or a herpesvirus vector.