雷帕霉素通过PU.1对树突状细胞DC-SIGN表达及生物学功能影响的研究  

Effect of rapamycin upon the expression of DC-SIGN on dendritic cells and the biological functions through the intracellular transcription factor PU.1

作　　者：陈静[1,2] 聂红[3] 蔡敏超[1] 张彦洁[1] 李晓[2] 陈楠[2] 许春娣[1] 周同[1,2] 

机构地区：[1]上海交通大学医学院附属瑞金医院肾脏科,上海200025 [2]上海交通大学医学院附属瑞金医院儿科,上海200025 [3]上海市交通大学医学院上海市免疫学研究所,上海200025

出　　处：《现代免疫学》2011年第6期467-472,共6页Current Immunology

基　　金：国家自然科学基金资助项目(30770999;81070567;81000163);上海市科委基础研究重点项目(09JC1409900)

摘　　要：探讨雷帕霉素(Rapamycin,Rapa)对树突状细胞(DC)表面DC-SIGN及其胞内转录因子PU.1基因表达,以及对DC功能的影响。体外培养人外周血单个核细胞来源的DC,结合PU.1基因特异性siRNA转染,给予不同浓度Rapa处理。采用流式细胞术及Western blot检测DC表面DC-SIGN表达变化;细胞迁移试验检测DC迁移能力;混合淋巴细胞反应检测DC刺激T细胞增殖能力。结果显示,Rapa呈剂量依赖性尤以10ng/ml时可下调DC-SIGN表达(P<0.01),同时可抑制DC胞内PU.1基因表达,以及DC迁移及刺激T细胞能力均受到明显抑制(P<0.01)。另发现,经siRNA沉默PU.1基因后,Rapa对DC-SIGN表达基本无影响。本研究表明,Rapa可通过抑制DC-SIGN表达,从而产生对DC迁移、刺激T细胞能力的调抑效应。这一效应可能与Rapa影响PU.1基因转录途径有关。To investigate the effect of rapamycin（Rapa） upon the expression of DC-SIGN on the surface of dendritic cells（DCs） and the genetic expression of the intracellular transcription factorthe PU.1 as well as the function of DCs.DCs derived from human peripheral blood mononuclear cells were cultured,and then treated with different concentrations of Rapa combined with PU.1-specific siRNA transfection.Flow cytometry and Western blotting were used to detect the expression of DC-SIGN on the surface of DC,while the cell transmigration test was used to detect the DC transmigrating ability,and the T lymphocyte proliferation was tested by means of mixed lymphocyte reaction（MLR）.The experimental results showed that Rapa could down-regulate the expression of DC-SIGN in a dose-dependent manner,especially in a dose of 10 ng/ml and also it could inhibit the genetic expression of PU.1,the transmigrating ability of DCs and to stimulate T lymphocyte proliferation.After the PU.1 gene was silenced by siRNA transfection,Rapa showed no effect on DC-SIGN expression virtually.From the results in the present study,it is evident that Rapa appears to show the down-regulating effect upon DC migration and the ability to stimulate T cell proliferation through its action to inhibit DC-SIGN expression and this effect may be associated with the influence on the PU.1 gene transcription pathway.

关 键 词：树突状细胞 固有免疫分子 转录因子 雷帕霉素 免疫调节 

分 类 号：R392.11[医药卫生—免疫学]