Heat-stable respiratory syncytial virus F protein oligomers and their use in immunological compositions

Abstract

Heat-stable oligomeric recombinant polypeptides, presenting at least one antigenic epitope of the pre-fusion Respiratory Syncytial Virus (RSV) F protein, comprising the RSV F protein ectodomain, functionally deleted in the HRB region, transmembrane and cytoplasmic domains replaced with a heterologous trimerization domain, and absent two functional multibasic furin cleavage sites, are useful as antigenic components in immunogenic compositions useful in methods of inducing an immune response and vaccinate against RSV infections.

The invention claimed is: 
     
       1. A heat-stable oligomeric recombinant polypeptide presenting at least one antigenic epitope of the pre-fusion Respiratory Syncytial Virus F protein, said polypeptide comprising
 the Respiratory Syncytial Virus F protein ectodomain, 
 from which the HRB region is deleted, and 
 from which the transmembrane and cytoplasmic domains are deleted and replaced withwhich Respiratory Syncytial Virus F protein ectodomain is extended with a heterologous trimerization domain,that replaces the deleted Respiratory Syncytial Virus F protein transmembrane and cytoplasmic domains; 
 and 
 wherein the two multibasic furin cleavage sites in said Respiratory Syncytial Virus F protein ectodomain are mutated by the substitution of all lysine arginine residues in said sites with arginine lysine residues, thereby rendering said furin cleavage sites defective. 
 
     
     
       2. The heat-stable oligomeric recombinant polypeptide according to  claim 1 , wherein said heterologous trimerization domain is a GCN4 leucine zipper trimerization motif. 
     
     
       3. The heat-stable oligomeric recombinant polypeptide according to  claim 1 , wherein said deletion of the HRB region comprises a deletion of the HRB wherein the HBR HRB comprises the sequence of SEQ ID NO: 10. 
     
     
       4. The heat-stable oligomeric recombinant polypeptide according to  claim 1 , further comprising a LysM peptidoglycan binding domain linked to the carboxy-terminal end of said trimerization domain. 
     
     
       5. The heat-stable oligomeric recombinant polypeptide according to  claim 1 , further comprising a triple Strep-tag. 
     
     
       6. The heat-stable oligomeric recombinant polypeptide according to  claim 1 , wherein said ectodomain is a soluble ectodomain. 
     
     
       7. The heat-stable oligomeric recombinant polypeptide according to  claim 1 , wherein said antigenic epitope is recognized by a pre-fusion specific monoclonal antibody AM22 or D25, or AM22 and D25. 
     
     
       8. An immunogenic composition comprising the oligomeric recombinant polypeptide of  claim 1 . 
     
     
       9. An immunogenic composition according to  claim 8 , further comprising an adjuvant. 
     
     
       10. An immunogenic composition according to  claim 8 , in wherein said oligomeric recombinant polypeptide is bound, covalently or non-covalently, to a carrier particle. 
     
     
       11. An immunogenic composition according to  claim 10 , wherein said carrier particle is a bacterium-like particle. 
     
     
       12. A recombinant expression vector comprising a nucleotide sequence encoding the polypeptide forming the heat-stable oligomer of  claim 1 . 
     
     
       13. A method of inducing an immune response in a subject to RSV comprising administering to said subject an immunogenic composition according to  claim 8 . 
     
     
       14. A heat-stable oligomeric recombinant polypeptide presenting at least one antigenic epitope of the pre-fusion Respiratory Syncytial Virus F protein, said polypeptide comprising
 the Respiratory Syncytial Virus F protein ectodomain, which is extended with a heterologous trimerization domain that replaces the deleted Respiratory Syncytial Virus F protein transmembrane and cytoplasmic domains; and   wherein the two multibasic furin cleavage sites in said Respiratory Syncytial Virus F protein ectodomain are mutated by the substitution of all arginine residues in said sites with lysine residues, thereby rendering said furin cleavage sites defective.   
     
     
       15. The heat-stable oligomeric recombinant polypeptide according to claim 14, wherein said heterologous trimerization domain is a GCN4 leucine zipper trimerization motif. 
     
     
       16. The heat-stable oligomeric recombinant polypeptide according to claim 14, further comprising a LysM peptidoglycan binding domain linked to the carboxy-terminal end of said trimerization domain. 
     
     
       17. The heat-stable oligomeric recombinant polypeptide according to claim 14, further comprising a triple Strep-tag. 
     
     
       18. The heat-stable oligomeric recombinant polypeptide according to claim 14, wherein said ectodomain is a soluble ectodomain. 
     
     
       19. The heat-stable oligomeric recombinant polypeptide according to claim 14, wherein said antigenic epitope is recognized by a pre-fusion specific monoclonal antibody AM22 or D25, or AM22 and D25. 
     
     
       20. An immunogenic composition comprising the oligomeric recombinant polypeptide of claim 14. 
     
     
       21. An immunogenic composition according to claim 20, further comprising an adjuvant. 
     
     
       22. An immunogenic composition according to claim 20, in wherein said oligomeric recombinant polypeptide is bound, covalently or non-covalently, to a carrier particle. 
     
     
       23. An immunogenic composition according to claim 22, wherein said carrier particle is a bacterium-like particle. 
     
     
       24. A recombinant expression vector comprising a nucleotide sequence encoding the polypeptide forming the heat-stable oligomer of claim 14. 
     
     
       25. A method of inducing an immune response in a subject to RSV comprising administering to said subject an immunogenic composition according to claim 20.