Development of T-cell lines expressing functional HIV-1 envelope glycoproteins for evaluation of immune responses in HIV-infected individuals.
The human T-lymphoid cell line, CEM, was transfected with gp 160 cDNA of human immunodeficiency virus type 1 (HIV-1)pm213. Three clones expressing the envelope glycoproteins (env), designated CEM-213env1, -env4, and -env7, were isolated. These clones expressed high levels of surface gp41 and gp120, as demonstrated by flow cytometry with anti-HIV env monoclonal antibodies. Processing and function of env was shown by induction of syncytia with CD4-expressing HeLa cells and by immunoblot analysis. The env expression resulted in specific down-regulation of surface CD4 levels, supporting the role of HIV env in CD4 modulation. Furthermore, serum samples from nine of nine HIV-1-infected individuals bound specifically to the env-expressing transfectants, substantiating the presence of conserved antigenic determinants. These sera also mediated antibody-dependent cellular cytotoxicity (ADCC) of the env-expressing cell lines. The env-expressing cell lines provide a relevant, safe, and practical model for qualitative and quantitative analysis of humoral and cellular immune responses and their role in HIV-1 pathogenesis and therapy.
Keler
T
T
Medarex, Inc., Annandale, New Jersey 08801, USA.
Li
H
H
Cloyd
M W
MW
Vitale
L A
LA
Deo
Y M
YM
D015153:Blotting, Western; D015704:CD4 Antigens / Q000378:metabolism; D002478:Cells, Cultured; D002999:Clone Cells / Q000276:immunology / Q000821:virology; D003602:Cytotoxicity, Immunologic; D015536:Down-Regulation; D000939:Epitopes / Q000276:immunology; D005434:Flow Cytometry; D015967:Gene Expression Regulation, Viral; D015726:Giant Cells / Q000821:virology; D015699:HIV Envelope Protein gp120 / Q000096:biosynthesis* / Q000276:immunology; D019302:HIV Envelope Protein gp160 / Q000096:biosynthesis* / Q000235:genetics / Q000276:immunology; D015700:HIV Envelope Protein gp41 / Q000096:biosynthesis* / Q000276:immunology; D015658:HIV Infections / Q000276:immunology / Q000821:virology*; D015497:HIV-1; D006367:HeLa Cells / Q000821:virology; D006801:Humans; D013601:T-Lymphocytes / Q000276:immunology / Q000821:virology*; D014162:Transfection
D015704:CD4 Antigens; D000939:Epitopes; D015699:HIV Envelope Protein gp120; D019302:HIV Envelope Protein gp160; D015700:HIV Envelope Protein gp41