甘露聚糖结合凝集素诱导DC成熟机制分析  

The mechanism of DC maturation induced by MBL

作　　者：王凡平[1] 王明永[1] 杨建斌[2] 赵东方[2] 王红坡[2] 邵峰[3] 孙瑞利[1] 郭庆合[1] 凌明智[1] 赵永新[3] 宋士军[3] 郭继强[1] 

机构地区：[1]新乡医学院医学检验系,新乡453003 [2]新乡医学院第一附属医院检验科,新乡453100 [3]新乡医学院第三附属医院检验科,新乡453003

出　　处：《中国免疫学杂志》2012年第12期1059-1062,1077,共5页Chinese Journal of Immunology

基　　金：国家自然科学基金资助项目(No.31100645);河南省科技攻关计划项目(No.102102310154);河南省高校科技创新人才项目(No.2012HASTIT024);河南省高校青年教师骨干项目(No.2010GGJS-117);河南省教育厅科学技术研究重点项目(No.12A320017)资助项目

摘　　要：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。Objective：To investigate the mechanism of mannan-binding lectin（MBL） on the differentiation and maturation of human peripheral blood monocyte-derived dendritic cells（DCs）.Methods：The monocytes were prepared from the blood of healthy adult volunteers.The immature dendritic cells（imDCs） were induced by 5-day-culture in medium supplemented with rhGM-CSF and rhIL-4.The expressions of CD83,CD86 and MHC-DR on DCs were analyzed by FACS.The interaction of MBL with imDCs was also investigated by FACS,and the levels of toll-like receptor 2（TLR2） and TLR4 mRNA expressions on these cells were determined by RT-PCR.Electrophoretic Mobility Shift Assay（EMSA） and Western blot was used to detect MBL-induced NF-κB activity and translocation in imDCs.Results：The expressions of CD83,CD86 and MHC-DR on the MoDCs were up-regulated by MBL,and that MBL could bind to imDCs directly in a Ca2＋-dependent manner.The mRNA expressions of TLR2 and TLR4 were down-regulated by MBL.MBL increased NF-κB activity and translocation in imDCs.Conclusions：MBL may affect the expressions of toll-like receptors and NF-κB activity in imDCs by binding to imDCs directly,suggesting that MBL can regulation DC maturation through ligand-binding.All these revealed some signaling pathways of MBL regulation of DC maturation.

关 键 词：甘露聚糖结合凝集素 树突状细胞 TOLL样受体 NF-ΚB 免疫调节 

分 类 号：R392.1[医药卫生—免疫学]