title: Stable expression of the transfected HIV-1 env gene in a human B cell line: characterization of gp120-expressing clones and immunobiological studies.
abstract: The env gene of human immunodeficiency virus type 1 (HIV-1) encodes gp 120/41 which plays an important role in the viral infection process and pathogenesis. The surface glycoprotein gp120 is a candidate molecule for the development of a subunit vaccine against HIV-1-induced acquired immunodeficiency syndrome (AIDS). However, thorough studies on the immunobiology of this molecule are hampered by the lack of a suitable model. With this background in mind, and in order to learn more on anti-gp120 cellular immunity, we attempted to develop gp120-expressing human cell clones. Thus by transfecting a human lymphoid cell line of B lineage (Raji), which is known to be resistant to the natural killer cell activity, with an expression vector encoding the envelope and vpu, we established three clones that stably express gp120/41 and vpu. The surface glycoprotein gp 120 is also expressed on the cell surface of these clones. The transfected cells from syncytia with CD4+ human cell lines as well as with peripheral blood mononuclear cells (PBMC) leading to the death of the fused cells. This observation represents additional evidence for the eventual depletion of CD4+ viral targets that fuse with adjacent HIV-infected, gp 120-expressing cells. The latent Epstein-Barr virus genome present in the transfected cells, was not induced to express the lytic cycle antigens. The densities of the surface expression of a number of molecules examined remained unchanged in the transfected cells except for the surface IgM, which increased significantly (P < 0.05) in two clones. One of the clones exhibited a significantly (P < 0.05) reduced proliferation rate as compared to the other clones. The transfected cells of all the three clones showed a significantly (P < 0.01) increased susceptibility to lysis by the PBMC from normal, healthy individuals in a 16-hr 51Cr-release assay. This is the first report of the MHC- and antibody-independent lysis of human cells transfected with the HIV-1 surface glycoprotein. The transfected cells also served as targets in a gp120-specific antibody-dependent cellular cytotoxicity assay. We anticipate that the present model will prove very useful for studying the gp120-specific immune responses in HIV-infected individuals.
authors.affiliation: Laboratory of Immunovirology, Sainte-Justine Hospital, Montreal, Qc., Canada.
authors.forename: A
authors.initials: A
authors.lastname: Ahmad
authors.forename: A
authors.initials: A
authors.lastname: Ladha
authors.forename: E A
authors.initials: EA
authors.lastname: Cohen
authors.forename: J
authors.initials: J
authors.lastname: Menezes
mesh_terms: D000163:Acquired Immunodeficiency Syndrome / Q000276:immunology*; D000920:Antibody-Dependent Cell Cytotoxicity; D001402:B-Lymphocytes; D001483:Base Sequence; D004261:DNA Replication; D015752:Genes, env; D005822:Genetic Vectors; D015699:HIV Envelope Protein gp120 / Q000096:biosynthesis / Q000235:genetics* / Q000276:immunology; D015700:HIV Envelope Protein gp41 / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology; D015497:HIV-1 / Q000235:genetics*; D006801:Humans; D015151:Immunoblotting; D007694:Killer Cells, Natural / Q000276:immunology*; D007700:Kinetics; D008715:Methionine / Q000378:metabolism; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides; D016133:Polymerase Chain Reaction; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000276:immunology; D013936:Thymidine / Q000378:metabolism; D014162:Transfection; D014407:Tumor Cells, Cultured
chemical_list: D015699:HIV Envelope Protein gp120; D015700:HIV Envelope Protein gp41; D009838:Oligodeoxyribonucleotides; D011994:Recombinant Proteins; D008715:Methionine; D013936:Thymidine