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Subpopulations and cytokine expression of nave and memory T cells in normal human PBMCs

LIU Jie, WU Chang-you Department of Immunology, Basic Medical College, Sun Yat-sen University, Key Laboratory of Tropical Disease Control of Ministry of Education, Guangzhou 510080, China  
AIM: To determine subpopulations and cytokine expression of nave and memory T cells by polychromatic flow cytometry (PFC). METHODS: PBMCs were separated from normal human peripheral blood and stimulated by superantigen (SEB). After incubation for 5 hours, the cells were fixed, permeablized and stained with multiplecolor-labeled mAbs for cell surface and intracellular cytokines. Then they were detected by polychromatic flow cytometry and the data were analyzed by Flow Jo software. RESULTS: Based on the expression of CD45RO molecules, CD4~+ and CD8~+ T cells were classified as nave and memory cells, which were further devided into several subsets according to the expression of CD62L and CCR7. When PBMCs were stimulated by SEB for 5 hours, CD45RO~+ and CD45RO~-CD4~+ or CD8~+ T cells expressed IL-2, IFN-γ and TNF-α. Further analysis indicated that CD62L~ lo CCR7~ lo and CCR7~+ T cells but not CD62L~ hi and CD62L~ hi CCR7~+ T cells expressed cytokines. In addition, the percentage of cytokine expression in CD62L~ lo CCR7~ lo subsets was markedly higher than that in CCR7~+ subsets. CONCLUSION: It is not scientific to identify nave and memory cells in human T cells only based on the expression of CD45RO. Accurate determination of nave, effector and memory cell populations can be achieved not only based on the expression of CD45RO but also according to the expression of CD62L.
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