Rapid screening for Mamu-A1-positive rhesus macaques using a SIVmac Gag peptide-specific cytotoxic T-lymphocyte assay.
As part of an ongoing vaccine study using peptide immunogens designed to stimulate simian immunodeficiency virus (SIV)mac-specific cytotoxic T lymphocytes (CTL) it was necessary to identify rhesus macaques within our colony bearing the Mamu-A1 major histocompatibility complex (MHC) class I haplotype. Peripheral blood mononuclear cells (PBMC) from individual monkeys were analysed by immunoelectrofocusing for the presence of a band corresponding to the Mamu-A1 molecule. In addition, PBMC were pulsed with the SIVmac Gag peptide 11 (against which CTL are Mamu-A1 restricted) and analysed for susceptibility to lysis by peptide 11-specific CTL. PBMC from all of the rhesus macaques shown to be Mamu-A1 positive by immunoelectrofocusing were highly sensitive to lysis by the peptide 11-specific CTL. A total of 46% (16 from 35) of the rhesus macaques originating from India were found to be Mamu-A1 positive, whereas none of the Chinese rhesus (0 from 37) macaques possessed this haplotype. Once a peptide-specific CTL is established, screening by CTL assay offers a faster, reliable and more relevant alternative to immunoelectrofocusing for selecting monkeys for use in vaccination trials.
Vogel
T
T
Paul-Ehrlich Institute, Langen, Germany.
Norley
S
S
Beer
B
B
Kurth
R
R
D000595:Amino Acid Sequence; D000818:Animals; D002460:Cell Line; D002478:Cells, Cultured; D003602:Cytotoxicity, Immunologic; D015686:Gene Products, env / Q000276:immunology*; D006239:Haplotypes; D015395:Histocompatibility Antigens Class I / Q000097:blood*; D006650:Histocompatibility Testing / Q000379:methods; D007525:Isoelectric Focusing; D007963:Leukocytes, Mononuclear / Q000276:immunology; D008253:Macaca mulatta; D008969:Molecular Sequence Data; D015682:Retroviridae Proteins, Oncogenic / Q000276:immunology*; D015302:Simian Immunodeficiency Virus / Q000276:immunology*; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology*; D014760:Viral Fusion Proteins / Q000276:immunology*
D015686:Gene Products, env; D015395:Histocompatibility Antigens Class I; D015682:Retroviridae Proteins, Oncogenic; D014760:Viral Fusion Proteins; C059580:transmembrane protein, Simian immunodeficiency virus