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目的 体外诱导和扩增树突状细胞 (DC)并分析其对自身T细胞的激发作用。方法 正常人外周血单个核细胞经贴壁去除悬浮细胞 ,加入细胞因子 (IL - 4、GM -CSF和TNF -α)培养 8天。用流式细胞仪分析细胞表型 ,用ELISA测定培养上清中IL - 12的含量 ,将诱导的树突状细胞与自身T细胞混合培养用3H -TdR法测定细胞的增殖指数。结果 正常人外周血诱导的树突状细胞高表达分化抗原CD1a(89.1% )、CD4 0 (99.8% )、CD80 (95 .1% )、CD83(4 5 .7% )、HLA -DR(97.6 % ) ,同时所获的DC能分泌IL - 12和有效地激活自身T细胞 ,并促其增殖。结论 正常人外周血的单个核细胞经细胞因子的序贯培养 ,可获得高纯度、功能性的DC细胞
Objective To induce and amplify dendritic cells (DCs) in vitro and analyze their effects on their own T cells. Methods Normal human peripheral blood mononuclear cells were suspended in adherent cells and cultured with cytokines (IL - 4, GM - CSF and TNF - α) for 8 days. The cell phenotype was analyzed by flow cytometry. The content of IL - 12 in culture supernatant was determined by ELISA. The induced dendritic cells were mixed with their own T cells and the proliferation index was determined by 3H - TdR method. Results The dendritic cells induced by peripheral blood of normal people were highly expressed differentiation antigen CD1a (89.1%), CD4 0 (99.8%), CD80 (95.1%), CD83 %). At the same time, the DCs secreted IL - 12 and effectively activated their own T cells and promoted their proliferation. Conclusion The normal human peripheral blood mononuclear cells by sequential cytokines can be obtained high purity, functional DC cells