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机构地区:[1]中山大学基础医学院免疫学教研室,广东广州510080
出 处:《细胞与分子免疫学杂志》2007年第6期491-494,共4页Chinese Journal of Cellular and Molecular Immunology
基 金:国家重点基础研究发展规划(973)资助项目(2001CB510007);广东省自然科学基金团队项目(05200176)
摘 要:目的:探讨CD154+细胞的表型特征与初始和记忆CD4+ T细胞以及与细胞因子表达之间的关系。方法:正常人外周血单个核细胞(PBMC),经抗CD3+抗CD28单克隆抗体(mAb)刺激培养后,利用多种抗细胞表面分子和抗细胞因子以及抗CD154抗体进行标记,用流式细胞术检测。结果:体外刺激PBMC6h后,CD154表达于CD4+ T细胞,而不表达于CD8+T细胞。对比分析CD4+CD154+ T和CD4+CD154- T细胞上CD45RA(初始)和CD45RO(记忆)表面分子的结果表明,大多数CD4+CD154+ T细胞为记忆细胞。当利用抗CD3或抗CD3+抗CD28mAb刺激后,分泌IFN-γ、IL-2和TNF-α的CD4+ T细胞均为CD4+CD154+ T细胞,而且单个细胞可以同时分泌两种以上细胞因子。进一步研究表明,CD4+CD154+ T细胞低表达或不表达CD25,不表达CD62L,50%左右的细胞表达CCR7。结论:体外短时间刺激PBMC,经过对细胞表型和细胞因子表达的研究,表明CD154分子结合其他表面标记或细胞因子的表达可以用于鉴别初始和记忆CD4+ T细胞。AIM: To investigate the correlation between CD154^+ T cells, naive/memory CD4^+ T cell markers and cytokine expression. METHODS: PBMC from normal persons were isolated, stained by fluorescence-conjugated monoclonal antibodies of surface markers, intracellular cytokines and CD154, and analyzed at the single cell level by flow cytometer. RESULTS: After stimulted in vitro for six hours, CD154 were expressed on CD4^+ T cells, rather than on CD8^+ T cells. Analysis of the expression of CD45RA ( marker for naive T ceils) and CD45RO (marker for memory T cells) on CD154^+ T cells indicated that the majority of CD4^+ CD154^+ T cells were memory T cells. When stimulated by antiCD3 or antiCD3 + antiCD28, the cells that produced IFN-γ IL-2 and TNF-α were CD4^+ CD154^+ and a single cell produced several cytokines simultaneously. In addition, CD4^+ CD154^+ T cells had low level or no expression of CD25 and they didn't express CD62L, but approximately fifty percent of the cells expressed CCR7. CONCLUSION: After stimulated in vitro for a short time and through further phenotypic and cytokine analysis, CD154, together with other cell surface markers and cytokines, can be used to discriminate between naive and memory CD4^+ T cells.
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