Synthesis and immunogenicity of the rotavirus major capsid antigen using a baculovirus expression system.
Rotaviruses are the major pathogens that cause life-threatening diarrhea in young children and animals. We inserted a simian rotavirus SA11 gene 6 cDNA into the genome of the baculovirus Autographa californica nuclear polyhedrosis virus adjacent to the strong polyhedrin promoter. The major capsid antigen (VP6) was expressed in high yields (20 to 150 micrograms/10(6) cells) when Spodoptera frugiperda cells were infected with baculovirus recombinants containing SA11 gene 6 inserts. Reactivity with monospecific polyclonal and monoclonal antibodies suggested that VP6, expressed intracellularly or found in the media, maintained native antigenic determinants. VP6 purified from the media from infected cells also possessed a native oligomeric structure, was immunogenic in guinea pigs, and was able to spontaneously assemble into morphologic subunits. Antisera from immunized guinea pigs failed to neutralize virus in plaque reduction assays, but detected homologous and heterologous rotavirus strains when tested by immunofluorescence, immunoprecipitation, and enzyme-linked immunosorbent assays.
D000914:Antibodies, Viral / Q000276:immunology; D000956:Antigens, Viral / Q000235:genetics*; D002213:Capsid / Q000235:genetics* / Q000276:immunology; D003001:Cloning, Molecular; D005786:Gene Expression Regulation; D005822:Genetic Vectors; D007304:Insect Viruses / Q000235:genetics*; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000276:immunology; D012401:Rotavirus / Q000235:genetics* / Q000276:immunology
D000914:Antibodies, Viral; D000956:Antigens, Viral; D011994:Recombinant Proteins