目的:探讨肿瘤坏死因子受体相关因子6(TRAF6)在不同成熟度树突状细胞(DCs)中的表达及其对DCs免疫刺激活性的影响.方法:重组人粒细胞-巨噬细胞集落刺激因子(rhGM-CSF)和重组人白细胞介素4(IL-4)体外诱导的人外周血单个核细胞来源的DCs,经TNF-α、LPS和细胞因子鸡尾酒法诱导成熟,分别以流式细胞术、ELISA和混合淋巴细胞反应(MLR)检测DCs的表面标志、IL-12分泌和刺激淋巴细胞增殖能力.Western blotting和real-time PCR分别检测各组TRAF6蛋白水平及mRNA的表达.结果:各组DCs均表达TRAF6,成熟状态最佳的鸡尾酒诱导下DCs的TRAF6蛋白表达最高.高表达TRAF6的DCs可促进IL-12的分泌,增强刺激淋巴细胞增殖的能力.结论:TRAF6是调节DCs激活和成熟的关键因素.%AIM: To investigate the immunological effect of tumor necrosis factor receptor - associated factor 6 ( TRAF6 ) on the maturation of dendritic cells in vitro.METHODS: The human peripheral blood mononuclear cell ( PBMC ) - derived dendritic cells ( DCs ), induced by recombined human granulocyte macrophage colony - stimulating factor ( GM - CSF ) and interleukin ( IL ) - 4, were stimulated to mature with TNF - α, LPS or cocktail of cytokines ( TNF - α, IL - 6, IL - 1 β and PGE2 ).The cell surface markers, T - cell stimulatory proliferation capacity and IL - 12 p40 production of the DCs were determined by the methods of flow cytometry, ELISA and mixed lymphocyte reation ( MLR ), respectively.The mRNA expression of TRAF6 was detected by real - time PCR.RESULTS: The expression of TRAF6 was observed in all groups, which in cocktail group was the highest in the DCs with optimum state of maturation.Furthermore, TRAF6 enhanced the production of IL - 12 and the ability of T - cell stimulation of mature DCs.CONCLUSION: TRAF6 might play an important role in inducing the maturation of human PBMC - derived DCs.
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