The rhesus rotavirus outer capsid protein VP4 functions as a hemagglutinin and is antigenically conserved when expressed by a baculovirus recombinant.
Rhesus rotavirus (RRV) gene 4 was cloned into lambda bacteriophage, inserted into a polyhedrin promoter shuttle plasmid, and expressed in Sf9 cells by a recombinant baculovirus. The baculovirus-expressed VP4 protein made up approximately 5% of the Spodoptera frugiperda-infected cell protein. Monoclonal antibodies that neutralize the virus bound to the expressed VP4 polypeptide, indicating that the expressed VP4 protein was antigenically indistinguishable from viral VP4. In addition, we have determined that the baculovirus-expressed VP4 protein bound to erythrocytes and functions as the RRV hemagglutinin. The endogenous hemagglutinating activity of the VP4 protein, like the virus, was inhibited by guinea pig antirotavirus hyperimmune serum and by VP4-specific neutralizing monoclonal antibodies. The human erythrocyte protein, glycophorin, also inhibited hemagglutination by RRV or the expressed VP4 protein and appears to be the rotavirus erythrocyte receptor. The baculovirus-expressed VP4 protein was conserved functionally and antigenically in the absence of other outer or inner capsid rotavirus components and represents a logical candidate for future immunological studies.
Mackow
E R
ER
Department of Medical Microbiology and Medicine, Stanford University, California 94305.
Barnett
J W
JW
Chan
H
H
Greenberg
H B
HB
D000818:Animals; D000956:Antigens, Viral / Q000235:genetics*; D002213:Capsid / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D004274:DNA, Recombinant; D005822:Genetic Vectors; D006021:Glycophorins / Q000378:metabolism; D006385:Hemagglutination Inhibition Tests; D006389:Hemagglutinins, Viral / Q000235:genetics* / Q000276:immunology; D007304:Insect Viruses / Q000235:genetics; D008253:Macaca mulatta; D011233:Precipitin Tests; D011991:Receptors, Virus / Q000378:metabolism; D012401:Rotavirus / Q000235:genetics* / Q000276:immunology
D000956:Antigens, Viral; D004274:DNA, Recombinant; D006021:Glycophorins; D006389:Hemagglutinins, Viral; D011991:Receptors, Virus