Mapping the fine specificity of a cytolytic T cell response to HIV-1 nef protein.
Epitope mapping of a MHC class I-restricted cytotoxic T cell response to nef, a regulatory protein of HIV, was performed with fresh PBMC from HIV-seropositive donors and target cells pulsed with a panel of overlapping peptides of the nef protein. These nef-specific CTL recognized a synthetic peptide of 10 residues derived from a nonamphipathic, highly conserved region of the nef protein in association with the HLA A3.1 molecule. Using human cell transfectants expressing mutations of the A3 molecule, we demonstrated that the amino acid at position 152 of the A3.1 molecule appears to be critical for detection of this response. Thus, rapid analysis of the epitopes of HIV proteins stimulating CTL responses can be achieved using a combination of fresh donor PBMC and target cells pulsed with synthesized peptides.
Laboratory of Immunoregulation, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD 20892.
S
S
Koenig
T R
TR
Fuerst
L V
LV
Wood
R M
RM
Woods
J A
JA
Suzich
G M
GM
Jones
V F
VF
de la Cruz
R T
RT
Davey
S
S
Venkatesan
B
B
Moss
D000595:Amino Acid Sequence; D003602:Cytotoxicity, Immunologic; D004252:DNA Mutational Analysis; D000939:Epitopes; D015693:Gene Products, nef / Q000235:genetics / Q000276:immunology*; D015483:HIV Antibodies / Q000276:immunology; D015488:HIV Antigens / Q000276:immunology*; D015497:HIV-1 / Q000276:immunology*; D015791:HLA-A3 Antigen / Q000276:immunology*; D006801:Humans; D007111:Immunity, Cellular; D008285:Major Histocompatibility Complex; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000276:immunology; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology*; D014162:Transfection; D054334:Viral Regulatory and Accessory Proteins / Q000276:immunology*; D054311:nef Gene Products, Human Immunodeficiency Virus
D000939:Epitopes; D015693:Gene Products, nef; D015483:HIV Antibodies; D015488:HIV Antigens; D015791:HLA-A3 Antigen; D011994:Recombinant Proteins; D054334:Viral Regulatory and Accessory Proteins; D054311:nef Gene Products, Human Immunodeficiency Virus