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Identification of a Neutralization-specific Antigen of a Calf Rotavirus

Department of Biological Sciences, University of Warwick, Coventry CV4 7AL, U.K.

Monospecific polyclonal antisera were raised in guinea-pigs against the calf rotavirus polypeptides VP1, VP2, VP3* + 4*, VP4.2, VP6, VP7.1, VP7.2 and VP10. All of the antisera gave a similar pattern of cytoplasmic immunofluorescence in rotavirus-infected cells, but spots of fluorescence of varying intensity with different sera were seen over the nucleus. Immune precipitation, using Staphylococcus aureus to collect immune complexes, showed that VP2 was precipitated by antiserum to VP2 (-VP2) and VP6 by -VP6. -VP7.1 and -VP7.2 both precipitated the same range of proteins from infected cells (VP7, VP7.1 and VP7.2) or from virions (VP7.1 and VP7.2). VP10, either from virions or infected cells, was not precipitated by -VP10. The only antiserum which efficiently neutralized infectivity was -VP7.2. There were low levels of neutralization with -VP10 (but the results varied from experiment to experiment) and traces with -VP6, -VP7.1 and the other antisera did not neutralize even though -VP7.1 agglutinated double-shelled particles as seen in immune electron microscopy to a greater extent than -VP7.2 Both VP7.1 and VP7.2 were shown to be glycoproteins by tunicamycin treatment of infected cells. Core particles only were agglutinated by -VP10. All the evidence leads us to conclude that there were major neutralizing antigenic determinants present on VP7.2, a minor component of the outer shell of the virion.

Keywords: calf rotavirus, monospecific antisera, neutralization antigen

Received 18 March 1982; accepted 13 May 1982.