Pre-fusion rsv f antigens

Abstract

The invention relates to pre-fusion RSV F protein and polypeptides that contain one or more amino acid mutations that stabilize the pre-fusion conformation or destabilize the post-fusion conformation. The invention also relates to methods for inducing an immune response to pre-fusion RSV F.

What is claimed is: 
     
         1 - 29 . (canceled) 
     
     
         30 . A pre-fusion respiratory syncytial virus (RSV) F polypeptide, wherein the HRA region (residues 137-239 of reference RSV F protein of SEQ ID NO:1) contains an introduced cysteine residue and the DIII region (residues 51-98 and 206-308 of reference RSV F protein of SEQ ID NO:1) contains an introduced cysteine residue, and a disulfide bond is formed between the introduced cysteine residue in the HRA region and the introduced cysteine residue in the DIII region that stabilizes the pre-fusion RSV F polypeptide. 
     
     
         31 . The pre-fusion RSV F polypeptide of  claim 30 , wherein said cysteine mutations are not more than about 10 Å away from each other. 
     
     
         32 . The pre-fusion RSV F polypeptide of  claim 30  wherein the prefusion RSV F polypeptide is a soluble ectodomain of RSV F. 
     
     
         33 . The pre-fusion RSV F polypeptide of  claim 30 , further comprising a heterologous oligomerization domain, an epitope, or a signal peptide. 
     
     
         34 . The pre-fusion RSV F polypeptide of  claim 33  comprising a heterologous oligomerization domain, wherein said heterologous oligomerization domain is a trimerization domain. 
     
     
         35 . The pre-fusion RSV F polypeptide of  claim 34  wherein the trimerization domain is selected from the group consisting of: influenza hemagglutinin, trimerizing sequence from bacteriophage T4 fibritin (“foldon”), SARS spike, HN gp41, NadA, modified GCN4, GCN4 or ATCase. 
     
     
         36 . The pre-fusion RSV F polypeptide of  claim 35 , wherein said heterologous oligomerization domain is the foldon. 
     
     
         37 . An immunogenic composition comprising the pre-fusion RSV F polypeptide of  claim 30 . 
     
     
         38 . The immunogenic composition of  claim 37 , further comprising an adjuvant. 
     
     
         39 . The immunogenic composition of  claim 38 , wherein the adjuvant is selected from the group consisting of: an aluminum salt, a squalene-in-water emulsion, a benzonaphthyridine compound, a phospholipid compound, a small molecule immunopotentiator and combinations of any of the foregoing. 
     
     
         40 . An isolated nucleic acid encoding the pre-fusion RSV F polypeptide of  claim 30 . 
     
     
         41 . The isolated nucleic acid of  claim 40 , which is a self-replicating RNA molecule. 
     
     
         42 . An immunogenic composition comprising the self-replicating RNA molecule of  claim 41  and a non-viral delivery system.