The VP8 fragment of VP4 is the rhesus rotavirus hemagglutinin.
The amino-terminal trypsin cleavage fragment of VP4, called VP8, was expressed from a recombinant baculovirus in Sf-9 cells. The baculovirus-expressed VP8 protein is antigenically conserved as demonstrated by its recognition by a library of neutralizing monoclonal antibodies. In Sf-9 cell sonicates, the expressed VP8 protein is capable of agglutinating human type O erythrocytes, indicating that the functionally intact rhesus rotavirus viral hemagglutinin is contained in the 247-amino acid VP8 trypsin cleavage fragment. Amino acid similarities between VP8 and the amino-terminal 282 amino acids of the reovirus sigma 1 protein suggests that the sigma 1 hemagglutination function resides within these amino-terminal amino acids as well. When the expressed VP8 protein was used to immunize mice, a broadly cross-reactive neutralizing antibody response was obtained. Antibodies elicited to the expressed VP8 protein neutralized viruses of serotypes 1-4 and 6 but not porcine strains OSU (st5) or Gottfried (st4). The neutralizing antibody response to VP8 appeared to be more cross-reactive than the immune response to expressed VP4 or to whole RRV virion. This suggests that subunit protein immunizations may broaden the neutralizing antibody immune responses to rotaviruses and enhance protective immunity to serotypically distinct strains.
Fiore
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Department of Medicine, Stanford University, California 94305.
Greenberg
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HB
Mackow
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D000818:Animals; D000914:Antibodies, Viral / Q000096:biosynthesis; D016367:Baculoviridae / Q000235:genetics; D002213:Capsid / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D036022:Capsid Proteins; D005260:Female; D015870:Gene Expression; D006389:Hemagglutinins, Viral / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D006868:Hydrolysis; D008253:Macaca mulatta; D051379:Mice; D008815:Mice, Inbred Strains; D009036:Moths / Q000382:microbiology; D012087:Reoviridae / Q000235:genetics; D012401:Rotavirus / Q000187:drug effects / Q000235:genetics / Q000276:immunology*; D014357:Trypsin / Q000494:pharmacology; D014614:Vaccines, Synthetic; D014764:Viral Proteins / Q000235:genetics / Q000276:immunology; D014765:Viral Vaccines
D000914:Antibodies, Viral; D036022:Capsid Proteins; D006389:Hemagglutinins, Viral; C060339:VP4 protein, Rotavirus; D014614:Vaccines, Synthetic; D014764:Viral Proteins; D014765:Viral Vaccines; C031728:sigma 1 protein, reovirus; D014357:Trypsin