1. Binding of human immunodeficiency virus type-1 (HIV-1) to partially purified membrane vesicles of lymphoblastoid cell line CEM.
- Author
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Benzair AB, Hirsch I, and Chermann JC
- Subjects
- Antibodies, Monoclonal, Binding, Competitive, Blotting, Western, CD4 Antigens analysis, Enzyme-Linked Immunosorbent Assay, HIV Core Protein p24 analysis, HIV Reverse Transcriptase, HIV-1 growth & development, Humans, Immunoblotting, RNA-Directed DNA Polymerase analysis, Sensitivity and Specificity, Tumor Cells, Cultured, CD4 Antigens metabolism, CD4-Positive T-Lymphocytes microbiology, Cell Membrane microbiology, HIV-1 metabolism, Virology methods
- Abstract
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).
- Published
- 1993
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