【摘要】 目的:探讨诱导树突状细胞成熟的最优方法。方法:以细胞因子GM-CSF和IL-4体外诱导人单核细胞来源的树突状细胞,分别采用CD40L、LPS、TNF-α、细胞因子鸡尾酒法(TNF-α、IL-6、IL-1β、PGE2)诱导成熟,24小时后收获DCs以流式细胞仪检测其成熟表型CD80、CD83、CD86、HLA-DR和FITC-Dextran的内吞能力,ELISA法检测其IL-12的分泌,MTT法检测其刺激淋巴细胞增殖活性。结果:CD40L、LPS、TNF-α、鸡尾酒法均可诱导DCs的成熟,其中以鸡尾酒法诱导成熟的效果最优,CD83的表达率为66.91%(P<0.05);成熟DCsFITC-Dextran的内吞能力明显下降;成熟DCsIL-12分泌量明显高于未成熟DCs,其中鸡尾酒法诱导成熟的DCs的IL-12分泌量最高,成熟的DCs有较强的刺激淋巴细胞增殖能力。结论:细胞因子鸡尾酒法是诱导DCs成熟的最佳方法。更多还原

【Abstract】 Objective:To investigate the most effective strategy for mature induction of dendritic cells.Methods:Human monocyte-derived dendritic cells were induced in the presence of cytokines GM-CSF and IL-4. On day 6, the immature DCs were pulsed with each of CD40L, LPS, TNF-α or a cocktail of cytokines(TNF-α, IL-6, IL-1β, PGE2). DCs were harvested after 24 h induction. The surface markers for maturation CD80,CD83,CD86 and HLA-DR were detected by FCM. FITC-Dextra endocytic activity was measured by FCM. IL-12 production was detected by ELISA. The capacity of DCs for T cell activation was detected by MTT assay.Results:CD40L,LPS,TNF-α and the cocktail of cytokines all could induce DCs’ maturation. The most effective scheme for induction of maturation was the cocktail of cytokines, and the expression rate of CD83 was up to 66.91%(P<0.05). The FITC-Dextran endocytic activity was sharply reduced after maturation. The amount of IL-12 production by mature DCs increased significantly, especially in induction of maturation using the cocktail cytokines. Mixed lymphocyte reaction demonstrated that the capacity for stimulating T cells sharply increased after DCs maturation.Conclusion:The most effective method for inducing DCs’ maturation was cocktail of cytokines.更多还原