title: Surface expression of the HIV-1 envelope proteins in env gene-transfected CD4-positive human T cell clones: characterization and killing by an antibody-dependent cellular cytotoxic mechanism.
abstract: The env gene of the human immunodeficiency virus-type 1 (HIV-1) was transfected in CEM-nkr, a human lymphoid cell line of T lineage that is resistant to the activity of natural killer cells, and for the first time, transfected T cell clones were established that stably express gp160 intracellularly and gp120 on the surface as demonstrated by radioimmunoprecipitation as well as by indirect membrane immunofluorescence. The regulatory protein vpu was not detected by radioimmunoprecipitation in these clones. The surface expression of gp120 without vpu in these clones provides direct evidence that gp160 is processed and cleaved (without vpu) in CD4+ cells. The CD4 antigens of these cells coprecipitated gp160; interestingly, no reduction of the surface CD4 expression (detectable by flow cytometric analysis of membrane immunofluorescence with OKT4) in the transfected cells was observed. However, decreased reactivity of the transfected clones with OKT4A was observed. The gp120-expressing cells did not form syncytia on coculture with other CD4+ human cell lines. These observations suggest the binding of gp120 to the surface CD4 antigen of the transfected cells. The transfected cells retained their resistance to the activity of the natural killer cells but showed a significant (p < 0.05) lysis when they were preincubated with AIDS patients' serum containing anti-gp120/41 antibodies. Thus, the expressed gp120/41 in these cells made them susceptible to killing by an antibody-dependent cellular cytotoxicity (ADCC) mechanism. To our knowledge, these are the first reported CD4+ T cell lines that stably express HIV envelope proteins. These cell lines would be useful as targets in exploring gp120/41-specific immune responses, especially in conducting gp120/41-specific ADCC studies in HIV-infected or gp120/41 (gp160)-vaccinated individuals.
authors.affiliation: Laboratory of Immunovirology, University of Montreal, Quebec, Canada.
authors.forename: A
authors.initials: A
authors.lastname: Ahmad
authors.forename: X A
authors.initials: XA
authors.lastname: Yao
authors.forename: J E
authors.initials: JE
authors.lastname: Tanner
authors.forename: E
authors.initials: E
authors.lastname: Cohen
authors.forename: J
authors.initials: J
authors.lastname: Menezes
mesh_terms: D000920:Antibody-Dependent Cell Cytotoxicity; D015704:CD4 Antigens / Q000276:immunology; D015496:CD4-Positive T-Lymphocytes / Q000382:microbiology*; D002460:Cell Line; D002999:Clone Cells; D005455:Fluorescent Antibody Technique; D015967:Gene Expression Regulation, Viral; D015686:Gene Products, env / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology; D015752:Genes, env; D015726:Giant Cells; D015699:HIV Envelope Protein gp120 / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology; D019302:HIV Envelope Protein gp160; D015497:HIV-1 / Q000235:genetics / Q000276:immunology*; D006801:Humans; D007694:Killer Cells, Natural / Q000276:immunology; D011498:Protein Precursors / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology; D015531:Radioimmunoprecipitation Assay; D014162:Transfection
chemical_list: D015704:CD4 Antigens; D015686:Gene Products, env; D015699:HIV Envelope Protein gp120; D019302:HIV Envelope Protein gp160; D011498:Protein Precursors