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目的:探讨甘露聚糖结合凝集素(mannan-binding lectin,MBL)对白假丝酵母菌(candida albicans,C.albicans)诱导的树突状细胞(dendritic cells,DC)成熟及分泌细胞因子的影响。方法:从健康成人外周血分离能粘附塑料的单核细胞(MNC),应用常规方法体外诱导未成熟DC(im DC),然后在有或无C.albicans和不同浓度(1-20mg/L)天然人MBL条件下继续培养2 d;用倒置显微镜观察DC的形态,以FACS分析DC的表型,用ELISA检测DC培养上清中TNF-α和IL-6的含量;FACS分析MBL与C.albicans及im DC的结合,Western blot分析IκBα磷酸化及p65/NF-κB细胞核移位。结果:ELISA检测发现,高浓度MBL(10-20 mg/L)能够下调C.albicans诱导人DC表面分子CD83和CD86表达,并抑制C.albicans诱导的TNF-α和IL-6分泌;FACS检测显示,MBL不仅能够与C.albicans结合而且能够与im DC结合;Western blot分析显示,MBL对C.albicans诱导的IκBα磷酸化及p65/NF-κB细胞核移位均有显著的抑制作用。结论:MBL能够以配体结合形式通过调节NF-κB信号途径抑制C.albicans诱导的DC成熟及细胞因子分泌,提示MBL能够在C.albicans诱导的免疫反应中起调控作用。
Objective: To investigate the effect of mannan-binding lectin (MBL) on the maturation and secretion of cytokines induced by candida albicans (C. albicans) in dendritic cells (DCs). METHODS: Plasticized mononuclear cells (MNCs) were isolated from peripheral blood of healthy adults and immature DCs were induced in vitro using conventional methods. ImDCs were then induced in vitro with or without C. albicans and different concentrations (1-20 mg / L ) Were cultured in natural human MBL for 2 days. The morphological changes of DCs were observed under an inverted microscope. The phenotypes of DCs were analyzed by FACS. The levels of TNF-α and IL-6 in DC culture supernatants were detected by ELISA. .albicans and im DC, Western blot analysis of IκBα phosphorylation and p65 / NF-κB nuclear translocation. Results: The results of ELISA showed that MBL (10-20 mg / L) could down-regulate the expression of CD83 and CD86 on human DCs and inhibit the secretion of TNF-α and IL-6 induced by C.albicans. The results showed that MBL could bind not only to C.albicans but also to im DC. Western blot analysis showed that MBL could significantly inhibit the phosphorylation of IκBα and the nuclear translocation of p65 / NF-κB induced by C.albicans. CONCLUSION: MBL can inhibit the maturation of DCs induced by C.albicans and the secretion of cytokines in a ligand-binding manner by regulating the NF-κB signaling pathway, suggesting that MBL may play a regulatory role in C.albicans-induced immune responses.