HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via CrossRef Citing Articles via Google Scholar Google Scholar Articles by Huggins, J. W. Articles by Linden, C. D. Search for Related Content PubMed PubMed Citation Articles by Huggins, J. W. Articles by Linden, C. D. Agricola Articles by Huggins, J. W. Articles by Linden, C. D.

Characterization of the Binding of the TC-83 Strain of Venezuelan Equine Encephalomyelitis Virus to BW-J-M, a Mouse Macrophage-like Cell Line

Department of Antiviral Studies, Department of Viral Pathogenesis
and¹ Immunology, Virology Division, U.S. Army Medical Research Institute of Infectious Diseases, Fort Detrick, Frederick, Maryland 21701, U.S.A.

The first step in virus replication is attachment of the virus to the host cell surface. To investigate this process, the binding of TC-83 (the attenuated strain of Venezuelan equine encephalomyelitis virus) to BW-J-M (a macrophage-like murine cell culture line) has been characterized. The binding of radiolabelled virus can be blocked by excess unlabelled virus and has a pH optimum in the physiological range. Binding is saturable; analysis using Scatchard plots or the computerized binding data analysis program, LIGAND, yielded estimates of a single class of 4 x 10⁵ binding sites per cell and an equilibrium binding constant of 2·0 ± 0·7 x 10¹² M^-1. Virus bound to the cell could be visualized by transmission electron microscopy and was localized primarily in coated regions of the membrane. Virus, bound under optimum conditions at 0 °C, was internalized upon warming and infected cells productively. Treatment of BW-J-M cells with low concentrations (1 to 10 µg/ml) of the proteolytic enzymes trypsin, Pronase or proteinase K caused a dose-dependent reduction in binding capacity. Trypsin-treated cells, upon return to culture, progressively regained their binding capacity within 24 h. As a further characterization of the virus binding site, several lectins were studied for their ability to inhibit TC-83 binding to BW-J-M cells. Canavalia ensiformis agglutinin, Glycine max agglutinin and Triticum vulgaris agglutinin were potent inhibitors of virus binding. This evidence suggests that TC-83 binds to a specific receptor on the BW-J-M cell and that the receptor may be glycoprotein.

Keywords: Venezuelan equine encephalitis, arbovirus, macrophage, binding

Received 23 February 1982; accepted 28 July 1982.