Neutralization of poliovirus by cell receptors expressed in insect cells.
To examine the interaction of the poliovirus receptor (PVR) with virus and the role of the PVR in virus entry, the PVR was expressed in insect cells. Poliovirus bound to insect cells infected with a recombinant baculovirus (AcPVR) carrying cDNA encoding the PVR. Antibodies raised against PVR expressed in bacteria immunoprecipitated a 67-kilodalton polypeptide from cytoplasmic extracts of AcPVR-infected cells. Treatment of AcPVR-infected cells with tunicamycin revealed that the PVR is a glycoprotein containing N-glycosidic linkages and that carbohydrate accounts for nearly 50% of its molecular weight as estimated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. When PVR was solubilized from AcPVR-infected insect cells and incubated with poliovirus, viral infectivity was neutralized. Sedimentation analysis revealed that irreversibly altered 135S particles were formed after incubation of poliovirus at 37 degrees C with solubilized extracts of AcPVR-infected insect cells. These results demonstrate that poliovirus eclipse may result from interaction with the cell receptor at neutral pH in the absence of membranes and suggest that soluble receptors may be effective antiviral agents against picornaviruses.
Kaplan
G
G
Department of Microbiology, College of Physicians and Surgeons, Columbia University, New York, New York 10032.
Freistadt
M S
MS
Racaniello
V R
VR
D000818:Animals; D002460:Cell Line; D004279:DNA, Viral / Q000235:genetics / Q000302:isolation & purification; D004797:Enzyme-Linked Immunosorbent Assay; D006367:HeLa Cells / Q000378:metabolism; D006801:Humans; D007304:Insect Viruses / Q000235:genetics*; D007313:Insecta; D009500:Neutralization Tests; D017955:Poliovirus / Q000502:physiology*; D011991:Receptors, Virus / Q000032:analysis / Q000235:genetics / Q000502:physiology*; D011994:Recombinant Proteins / Q000032:analysis / Q000378:metabolism; D014771:Virion / Q000502:physiology
D004279:DNA, Viral; D011991:Receptors, Virus; D011994:Recombinant Proteins