Detection of primary cytotoxic T lymphocytes specific for the envelope glycoprotein of HIV-1 by deletion of the env amino-terminal signal sequence.
A heterogenous population of envelope glycoprotein-specific cytotoxic effector cells are found in the peripheral blood of individuals infected with HIV-1, and in many cases env-specific lysis is not restricted by MHC molecules and is not blocked by antibody to CD3 (Rivière, Y. et al., J. Virol. 1989, 63:2270). In order to detect env-specific cytotoxic T lymphocytes (CTL) in fresh peripheral blood mononuclear cells of HIV-1-infected donors, a mutant env gene with deletion of the amino-terminal signal sequence was inserted into vaccinia virus. This deletion of the amino-terminal signal sequence was inserted into vaccinia virus. This deletion results in synthesis of an envelope protein that is not glycosylated and not expressed at the surface of infected cells. Target cells infected with this recombinant vaccinia virus are not lysed by antibody-mediated cellular cytotoxicity, but they are recognized by secondary CTL. Comparing lysis of target cells expressing gp160 of HIV-1 and the signal peptide deletion mutant, primary env-specific CTL were detected in some individuals infected with HIV-1.
McChesney
M
M
Département de Médecine, Institut Pasteur, Paris, France.
Tanneau
F
F
Regnault
A
A
Sansonetti
P
P
Montagnier
L
L
Kieny
M P
MP
Rivière
Y
Y
D000595:Amino Acid Sequence; D000920:Antibody-Dependent Cell Cytotoxicity; D000945:Antigens, Differentiation, T-Lymphocyte / Q000276:immunology; D001483:Base Sequence; D016827:CD8 Antigens; D004252:DNA Mutational Analysis; D015686:Gene Products, env / Q000235:genetics / Q000276:immunology*; D006678:HIV / Q000235:genetics / Q000276:immunology*; D015488:HIV Antigens / Q000276:immunology*; D015658:HIV Infections / Q000276:immunology*; D006801:Humans; D008285:Major Histocompatibility Complex; D008969:Molecular Sequence Data; D021382:Protein Sorting Signals; D011994:Recombinant Proteins; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology*; D014616:Vaccinia virus
D000945:Antigens, Differentiation, T-Lymphocyte; D016827:CD8 Antigens; D015686:Gene Products, env; D015488:HIV Antigens; D021382:Protein Sorting Signals; D011994:Recombinant Proteins