Early interactions of human herpesvirus 6 with lymphoid cells: role of membrane protein components and glycosaminoglycans in virus binding.
A microassay was developed to detect human herpesvirus 6 (HHV-6) binding to its cellular receptor using flow cytometry. Comparable results were obtained either by using HHV-6 preparations conjugated with fluorescein isothiocyanate or by indirect immunofluorescent labeling of membrane-bound virus using as primary antibody a monoclonal antibody specific for the HHV-6 gp60/110 envelope glycoprotein. Virus attachment to the plasma membrane was specific and saturable. As expected, among cell lines of various origin, maximum binding was detected on human T-lymphoid cells (HSB-2). Papain digestion of HSB-2 cells prevented HHV-6 attachment and reduced significantly virus infection, indicating the involvement of a protein-based receptor in the attachment step. After removal of the protease, virus receptors were resynthesized and their regeneration was prevented partially by cycloheximide, an inhibitor of protein synthesis. Unexpectedly, only high concentrations (mg/ml) of soluble heparan sulfate and heparin inhibited HHV-6 binding and infection. Under the same conditions, few micrograms (per ml) of heparin suppressed completely herpes simplex type 1 (HSV-1) attachment to the same cell line. Treatment of HSB-2 cells with heparitinase and heparinase, at doses that reduced significantly HSV-1 attachment, had little effect on HHV-6 binding to the cell membrane, indicating a different requirement of heparan sulfate-containing glycosaminoglycans for the two herpesviruses. These data suggest that protein components of the cellular membrane play an essential role in HHV-6 binding and infection while heparan sulfate-glycos-aminoglycans appear to be involved only partially in virus-receptor interaction.
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Istituto di Microbiologia, Università di Roma "La Sapienza," Roma, Italy.
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D000818:Animals; D050260:Carbohydrate Metabolism; D002241:Carbohydrates / Q000502:physiology; D002460:Cell Line; D002522:Chlorocebus aethiops; D004798:Enzymes / Q000378:metabolism; D005434:Flow Cytometry / Q000379:methods; D006025:Glycosaminoglycans / Q000378:metabolism / Q000502:physiology*; D015654:Herpesvirus 6, Human / Q000378:metabolism*; D006801:Humans; D008214:Lymphocytes / Q000378:metabolism / Q000821:virology*; D008565:Membrane Proteins / Q000378:metabolism / Q000502:physiology*; D011991:Receptors, Virus / Q000096:biosynthesis / Q000378:metabolism / Q000502:physiology*; D013997:Time Factors; D014407:Tumor Cells, Cultured; D014709:Vero Cells
D002241:Carbohydrates; D004798:Enzymes; D006025:Glycosaminoglycans; D008565:Membrane Proteins; D011991:Receptors, Virus