Characterisation of G serotype dependent non-antibody inhibitors of rotavirus in normal mouse serum.
Serotype specific (non-immunoglobulin) inhibitors of rotavirus have been identified in normal mouse serum obtained from BALB/c, CBA, and BL10 mice. Sialic acid was essential for the neutralising activity sera treated with the neuraminidase from Vibrio cholerae failed to neutralise rotavirus. G serotypes 4, 5, 7, 8, 9, and 10 were unaffected by the inhibitor(s) while G serotypes 1, 2, 6 and two G3 strains were neutralised to significant titres. Assessment of neutralisation of reassortants suggested that VP7 is the virus protein involved in the interaction although it remains possible that VP7 is influencing VP4 binding. Analysis of the sera by Western blot followed by virus overlay confirmed that binding is dependent on the presence of sialic acid. The human strain tested, Wa, bound to two (glyco) proteins (50 and 80 kDa) while the bovine strains tested, NCDV and UK bound to one (55 kDa) and two (36 and 55 kDa) proteins respectively. This indicates that while the bovine rotaviruses may bind to a common element, the human strain binds to clearly distinct proteins. We propose that these inhibitors interact with animal rotaviruses in a manner analogous to that by which they attach to target cells. The glycoprotein to which NCDV bound was purified and identified by N-terminal sequencing as murine alpha-1-anti-trypsin (MuAAT) and was confirmed to possess both neutralisation and anti-trypsin activity. Since MuAAT is known to possess only three N-linked glycans, identification and analysis of the actual virus-binding structure should now be possible.
Department of Microbiology and Immunology, University of Melbourne, Parkville, Victoria, Australia.
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Beisner
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Kool
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Marich
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Holmes
D000595:Amino Acid Sequence; D000818:Animals; D000914:Antibodies, Viral / Q000097:blood; D000956:Antigens, Viral; D000998:Antiviral Agents / Q000097:blood* / Q000235:genetics; D002213:Capsid / Q000276:immunology; D036022:Capsid Proteins; D002417:Cattle; D006801:Humans; D066298:In Vitro Techniques; D051379:Mice; D008807:Mice, Inbred BALB C; D008808:Mice, Inbred CBA; D008969:Molecular Sequence Data; D019158:N-Acetylneuraminic Acid / Q000097:blood; D009439:Neuraminidase / Q000494:pharmacology; D009500:Neutralization Tests; D012401:Rotavirus / Q000145:classification* / Q000276:immunology* / Q000472:pathogenicity; D017386:Sequence Homology, Amino Acid; D012703:Serotyping; D000515:alpha 1-Antitrypsin / Q000235:genetics
D000914:Antibodies, Viral; D000956:Antigens, Viral; D000998:Antiviral Agents; D036022:Capsid Proteins; C060339:VP4 protein, Rotavirus; C050779:VP7 protein, Rotavirus; D000515:alpha 1-Antitrypsin; D009439:Neuraminidase; D019158:N-Acetylneuraminic Acid