Characterization of T-cell subsets and T-cell receptor subgroups in pigtailed macaques using two- and three-color flow cytometry.
In order to characterize macaque T-lymphocyte subsets, we used a chromophore from a dinoflagellate, peridinin chlorophyll A protein (PerCP), which, like fluorescein isothiocyanate (FITC) and R-phycoerythrin (PE), can be excited by a 488-nm laser and emits light at 670 nm without spectral overlap with FITC and PE. Mouse monoclonal antibodies were conjugated with FITC, PE, and PerCP to detect CD4+ and CD8+ cells in macaque peripheral blood lymphocytes (PBL) subsets before and after activation and in nonactivated thymocytes. Resting and activated macaque blood CD4+ T-cells could be clearly delineated into discrete subsets with either CD28, CD45RA, or CD45RO as a second marker and CD26, CD29, CD44, or CD69 as a third marker. CD8+ cells were further subdivided by expression of similar combinations of markers. A subset of CD8+ CD28- T-cells in blood expressed the activation marker CD69, suggesting that they were already activated. Virtually all CD4+CD8+, CD4+CD8-, and CD4-CD8+ macaque thymocytes expressed CD2, CD3, and CD18 and not CD25, CD44, or CD45O, but macaque thymocyte subpopulations did differ in their expression of CD28 and CD29. The expression of T-cell receptor (TCR) subgroups on macaque PBL and thymocytes was analyzed before and after activation with staphylococcal enterotoxins (superantigens). The pattern of T-cell variable-region expression in macaques was similar to that seen in humans, with a high frequency of T cells expressing V beta 8. After superantigen stimulation, only minor changes in TCR V beta expression were detectable in PBL. A dramatic increase in V beta 8 expression was seen after stimulation of macaque thymus with staphylococcal enterotoxin D (SE-D), a minor increase after toxic shock syndrome toxin 1 (TSST-1) stimulation, and a simultaneous decrease in V beta 6 levels.
Regional Primate Research Center, University of Washington, Seattle 98195.
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I
Axberg
M J
MJ
Gale
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B
Afar
E A
EA
Clark
D000818:Animals; D016516:CD4-CD8 Ratio; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology; D002469:Cell Separation; D004768:Enterotoxins / Q000494:pharmacology; D005434:Flow Cytometry; D005456:Fluorescent Dyes; D005803:Genes, Immunoglobulin / Q000276:immunology; D006801:Humans; D008213:Lymphocyte Activation / Q000276:immunology; D008254:Macaca nemestrina; D011948:Receptors, Antigen, T-Cell / Q000276:immunology*; D013211:Staphylococcus aureus; D016176:T-Lymphocyte Subsets / Q000276:immunology*; D050378:T-Lymphocytes, Regulatory / Q000276:immunology
D004768:Enterotoxins; D005456:Fluorescent Dyes; D011948:Receptors, Antigen, T-Cell