Binding of human immunodeficiency virus type-1 (HIV-1) to partially purified membrane vesicles of lymphoblastoid cell line CEM.
Binding of human immunodeficiency virus type 1 (HIV-1) to membrane of its target cells was studied by a quantitative and non-isotopic method called the viral membrane trapping method (VMTM). Membranes prepared from the CD4 positive lymphoblastoid cell line CEM and adsorbed to a solid support retained the ability to bind HIV-1. Similar results were obtained by Western dot blot and ELISA modification of VMTM, when membrane fraction was bound to nitrocellulose or polystyrene, respectively. In ELISA modification, viral association with 1 microgram of membranes coated on 96-well microplate was linear within a range of 3.75 micrograms to 60 micrograms of p24gag protein. The use of anti-CD4 mAbs, OKT4A and 13B8.2, identified CD4 molecule as a major HIV-1 binding component of membrane fraction. The procedure will allow the study of virus binding to this and to other possible additional receptor(s).
Benzair
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AB
Unité de Recherches INSERM sur les Rétrovirus et Maladies Associées (U.322), Campus Universitaire de Luminy, Marseille, France.
Hirsch
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I
Chermann
J C
JC
D000911:Antibodies, Monoclonal; D001667:Binding, Competitive; D015153:Blotting, Western; D015704:CD4 Antigens / Q000032:analysis / Q000378:metabolism*; D015496:CD4-Positive T-Lymphocytes / Q000382:microbiology*; D002462:Cell Membrane / Q000382:microbiology*; D004797:Enzyme-Linked Immunosorbent Assay; D016655:HIV Core Protein p24 / Q000032:analysis; D054303:HIV Reverse Transcriptase; D015497:HIV-1 / Q000254:growth & development / Q000378:metabolism*; D006801:Humans; D015151:Immunoblotting; D012194:RNA-Directed DNA Polymerase / Q000032:analysis; D012680:Sensitivity and Specificity; D014407:Tumor Cells, Cultured; D014773:Virology / Q000379:methods*
D000911:Antibodies, Monoclonal; D015704:CD4 Antigens; D016655:HIV Core Protein p24; D054303:HIV Reverse Transcriptase; D012194:RNA-Directed DNA Polymerase