论文部分内容阅读
目的:探讨甘露聚糖结合凝集素(Mannan-binding lectin,MBL)诱导人外周血单核细胞(Mo)来源树突状细胞(DC)成熟的机制。方法:从健康成人外周血分离Mo,常规方法体外诱导未成熟DC(imDC),FACS分析DC成熟过程中MBL对CD分子表达的影响,进一步分析MBL与imDC的结合情况;RT-PCR分析DC表面模式识别分子Toll样受体-2(TLR2)和Toll样受体-4(TLR4)的表达;凝胶电泳迁移率法(EMSA)和Western blot分析NF-κB的活性及细胞核移位。结果:FCM分析表明,MBL能够使DC表面CD83、CD86及MHC分子表达升高;MBL能够以Ca2+浓度依赖关系与imDC细胞结合;RT-PCR结果表明MBL能够减少DC表面TLR2和TLR4的表达。EMSA和Western blot分析结果显示,MBL能够增强NF-κB活性及细胞核移位。结论:MBL可通过直接结合imDC表面分子来影响DC模式识别分子的表达并调节信号分子NF-κB活性,提示MBL能够通过配体结合调控DC分化成熟,揭示了MBL调节DC分化成熟有关信号途径。
AIM: To investigate the mechanism of Mannan-binding lectin (MBL) -induced dendritic cell (DC) maturation in human peripheral blood mononuclear cells (Mo). Methods: Mo was isolated from peripheral blood of healthy adults. ImDC was induced by conventional methods in vitro. FACS analysis of the effect of MBL on CD expression was performed by FACS. The binding of MBL to imDC was further analyzed. RT-PCR analysis of DCs The expression of TLR2 and TLR4 were detected by Western blotting. The activity and nuclear translocation of NF-κB were analyzed by EMSA and Western blot. Results: FCM analysis showed that MBL increased the expression of CD83, CD86 and MHC on DCs. MBL could bind to imDC cells in a Ca2 + concentration-dependent manner. RT-PCR results showed that MBL decreased the expression of TLR2 and TLR4 on DCs. EMSA and Western blot analysis showed that MBL enhanced NF-κB activity and nuclear translocation. CONCLUSION: MBL can directly affect the expression of DC pattern recognition molecule and regulate NF-κB activity by binding to imDC surface molecules, suggesting that MBL can regulate DC differentiation and maturation through ligand binding, revealing that MBL regulates the signaling pathways involved in DC maturation.