Flow cytometric method to demonstrate whether anti-HIV-1 agents inhibit virion binding to T4+ cells.
The first step in the replicative cycle of human immunodeficiency virus type 1 (HIV-1) is binding of the virions to the cellular CD4 receptor. This process may be considered as an important target for chemotherapeutic agents against acquired immune deficiency syndrome (AIDS). A method has now been devised whereby virion binding to the cell membrane was visualized by an indirect immunofluorescence assay using human anti-HIV-1 serum, rabbit anti-human-IG-F(ab')2-fluorescein isothiocyanate, and flow cytometry. Heparin, dextran sulfate, and pentosan polysulfate suppressed HIV-1 binding to MT-4 cells at concentrations that protected the cells against HIV-1 cytopathogenicity. Dextran and dermatan sulfate, two compounds that are inactive against HIV-1, had no inhibitory effect on the binding of HIV-1 to the cells. The potent and selective HIV-1 inhibitor azidothymidine (AZT) did not affect virus binding to the cells, whereas suramin partially blocked HIV-1 binding to the cells at concentrations that fully protected MT-4 cells against destruction of HIV-1. Our immunofluorescence assay thus demonstrated that suramin not only acts as an inhibitor of reverse transcriptase but also interferes with virus-cell binding. Also, Evans blue, an anionic dye structurally related to suramin, partially inhibited HIV-1 attachment to the cells. The present method permits a quantitative determination of the inhibitory effect of anti-HIV-1 agents on virion-cell binding.
Schols
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Rega Institute for Medical Research, University of Leuven, Belgium.
Baba
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Pauwels
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R
De Clercq
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E
D000945:Antigens, Differentiation, T-Lymphocyte / Q000032:analysis; D000998:Antiviral Agents / Q000494:pharmacology*; D003871:Dermatan Sulfate / Q000494:pharmacology; D016264:Dextran Sulfate; D003911:Dextrans / Q000494:pharmacology; D005434:Flow Cytometry; D015483:HIV Antibodies / Q000032:analysis; D015497:HIV-1 / Q000187:drug effects*; D006493:Heparin / Q000494:pharmacology; D006801:Humans; D010426:Pentosan Sulfuric Polyester / Q000494:pharmacology; D015728:Receptors, HIV; D011991:Receptors, Virus / Q000378:metabolism*; D013498:Suramin / Q000494:pharmacology; D013601:T-Lymphocytes / Q000276:immunology / Q000382:microbiology*; D014771:Virion / Q000378:metabolism*; D015215:Zidovudine / Q000494:pharmacology
D000945:Antigens, Differentiation, T-Lymphocyte; D000998:Antiviral Agents; D003911:Dextrans; D015483:HIV Antibodies; D015728:Receptors, HIV; D011991:Receptors, Virus; D003871:Dermatan Sulfate; D010426:Pentosan Sulfuric Polyester; D015215:Zidovudine; D013498:Suramin; D006493:Heparin; D016264:Dextran Sulfate