Flow microfluorometric analysis of peripheral blood mononuclear cells from nonhuman primates: Correlation of phenotype with immune function

Abstract

A panel of monoclonal antibody reagents has been identified that can be used for routine monitoring of subsets of peripheral blood mononuclear cells (PBMC) from Macaca mulatta (rhesus macaques), Macaca nemestrina (pig-tailed macaques), and Cercocebus atys (sooty mangabeys). The procedure uses fluorescein and phycoerythrin conjugates of the monoclonal antibodies in appropriate combinations, so that two-color microfluorometric analyses can be readily performed on as little as 1.2 ml of EDTA blood. PBMC from a total of 20 normal adult rhesus macaques, 21 normal adult pig-tailed macaques, 4 SIV- sooty mangabeys, and 16 SIV+ adult sooty mangabeys were analyzed with the panel of monoclonal reagents and flow microfluorometry. The mean frequency, absolute numbers, and range for each subset in these nonhuman primate species are described. Sooty mangabeys appeared markedly different from the other two primate species. The PBMCs from the mangabeys had a higher mean frequency and absolute number of total T cells, Leu-3a+/18- T cells, suppressor (Leu-2a+) T cells, which were HLA-DR+, and IL-2R+ cells. Functional helper, suppressor, natural killer (NK), lymphokine activated killer (LAK), and antigen-presenting cell studies were also performed to correlate phenotype with immune function. Data indicate that Leu-3a+ T cells (CD4+) and Leu-2a+ T cells (CD8+) in these primate species represent human equivalents of helper and suppressor T cells, respectively. NK and LAK effector cells in the rhesus and pig-tailed macaques appear to be predominantly Leu-19+. In contrast, Leu-2a+ cells appear to be the predominant NK and LAK effector cell in sooty mangabeys. These data provide a basis for routine evaluation of lymphocyte subsets in these nonhuman primate species, and provide a means to correlate phenotype with immune function.