Norovirus and sapovirus antigens

Abstract

Immunogenic compositions that elicit immune responses against Norovirus and Sapovirus antigens are described. In particular, the invention relates to polynucleotides encoding one or more capsid proteins or other immunogenic viral polypeptides from one or more strains of Norovirus and/or Sapovirus, coexpression of such immunogenic viral polypeptides with adjuvants, and methods of using the polynucleotides in applications including immunization and production of immunogenic viral polypeptides and viral-like particles (VLPs). Methods for producing Norovirus- or Sapovirus-derived multiple epitope fusion antigens or polyproteins and immunogenic compositions comprising one or more immunogenic polypeptides, polynucleotides, VLPs, and/or adjuvants are also described. The immunogenic compositions of the invention may also contain antigens other than Norovirus or Sapovirus antigens, including antigens that can be used in immunization against pathogens that cause diarrheal diseases, such as antigens derived from rotavirus.

1 . A method for producing Norovirus viral-like particles (VLPs), the method comprising:
 a) transforming a yeast cell with a polynucleotide comprising a coding sequence for a Norovirus VP1 or a particle forming polypeptide therefrom, and/or a polynucleotide comprising a coding sequence for a Norovirus VP2 or a particle forming polypeptide therefrom, wherein said polynucleotide is operably linked to control elements that direct transcription of said coding sequence in said yeast cell;   b) culturing the transformed yeast cell under conditions whereby VLPs are produced.   
     
     
         2 . The method of  claim 1 , wherein the yeast cell is transformed with a polynucleotide comprising a coding sequence for a Norovirus VP1 or a particle forming polypeptide therefrom and a polynucleotide comprising a coding sequence for a Norovirus VP2 or a particle forming polypeptide therefrom. 
     
     
         3 . The method of  claim 2 , wherein said polynucleotides are within the same expression vector. 
     
     
         4 . The method of  claim 2 , wherein said polynucleotides are within different expression vectors. 
     
     
         5 . The method of  claim 1 , wherein one or more of the polynucleotides is from more than one Norovirus isolate. 
     
     
         6 . The method of  claim 1 , further comprising transforming said yeast cell with one or more sequences encoding a structural protein from a Norovirus. 
     
     
         7 . The method of  claim 1 , wherein a mosaic VLP comprising capsid proteins from at least two viral strains of Norovirus is assembled. 
     
     
         8 . The method of  claim 1 , wherein the expression vector comprises an ADH2 promoter. 
     
     
         9 . The method of  claim 8 , wherein the promoter is a hybrid ADH2/GAPDH promoter. 
     
     
         10 . A VLP produced according to the method of  claim 1 . 
     
     
         11 . The VLP of  claim 10 , wherein said VLP comprises an amino acid sequence with at least 90% sequence identity to (i) the amino acid sequence of SEQ ID NO:3; (ii) the amino acid sequence of SEQ ID NO:4; and (iii) a particle forming polypeptide from (i) or (ii). 
     
     
         12 . The VLP of  claim 11 , wherein said VLP comprises the amino acid sequence of SEQ ID NO:3. 
     
     
         13 . The VLP of  claim 11 , wherein said VLP comprises the amino acid sequence of SEQ ID NO:4. 
     
     
         14 . The VLP of  claim 11 , wherein said VLP comprises the amino acid sequence of SEQ ID NO:3 and SEQ ID NO:4. 
     
     
         15 . A composition comprising the VLP of  claim 11  and a pharmaceutically acceptable excipient. 
     
     
         16 . A composition comprising the VLP of  claim 12  and a pharmaceutically acceptable excipient. 
     
     
         17 . A composition comprising the VLP of  claim 13  and a pharmaceutically acceptable excipient. 
     
     
         18 . A composition comprising the VLP of  claim 14  and a pharmaceutically acceptable excipient.