抑制小鼠骨髓树突状细胞RelB基因诱导免疫耐受的体外研究  

The effect of nuclear factor RelB suppression on the induction of immunological tolerance in vitro of mice bone marrow-derived dendritic cells

作　　者：包杰[1] 王前 郑磊[1] 熊石龙[1] 裘宇容[1] 

机构地区：[1]南方医科大学南方医院检验医学科,广州510515 [2]南方医院院办公室,广州510515

出　　处：《现代免疫学》2010年第6期488-492,共5页Current Immunology

基　　金：广东省自然科学基金资助项目(8451051501000580)

摘　　要：采用rmGM-CSF和rmIL-4联合诱导培养小鼠骨髓树突状细胞(bone marrow dendritic cells),经免疫磁珠方法纯化;利用慢病毒载体制备RelB shRNA慢病毒,与小鼠骨髓树突状细胞共培养,经流式细胞术观察树突状细胞表面分子MHCⅡ、CD86和CD40的表达;采用CD3+免疫磁珠分离纯化小鼠T细胞;分别采用四唑蓝(MTT)比色法和液相芯片法检测异基因T细胞增殖的程度以及Th1/Th2细胞因子的分泌水平。此外,实验设LPS-DC组、未处理组和LPS RNAi RelB DC组。发现核因子RelB抑制的树突状细胞表面分子MHCⅡ、CD86和CD40均低水平表达,显著低于LPS-DC表面分子的表达(P<0.05),且该DC经LPS刺激后(LPS RNAi RelB DC)表面上述3类分子的表达水平仍显著低于LPS-DC组(P<0.05),与未处理组(immature DC)表面分子表达水平相当。其刺激异基因小鼠T细胞增殖的能力和Th1/Th2分子的分泌水平与未成熟DC组相当。表明抑制核因子RelB的骨髓树突状细胞在体外具有诱导T细胞免疫耐受的能力,是一种新型的致耐受树突状细胞研究方法。Mouse bone marrow derived dendritic cells（DC） were cultured in RPMI 1640 with recombinant mouse granulocyte-macrophage colony-stimulating factor（rmGM-CSF） and interleukin-4（rmIL-4）,and purified by CD11c＋ microbead.RelB sh-RNA lentivirus were prepared by using kits of Invitrogen Corp,and co-cultured with mouse bone marrow derived dendritic cells,then surface molecules（MHCⅡ,CD86 and CD40） expression level were determined by flow cytometry（FCM）,and the mouse T cells were purified by CD3＋ microbeads.Then T cell proliferation capacity and Th cytokine detected by MTT approach,and by liquid chip approach respectively.Mature DC（that is,LPS-DC） worked as control group,at the same time,we also designed an immature DC and LPS RNAi RelB DC.We found that the expression levels of co-stimulatory molecules（CD86 and CD40） and MHCⅡ class molecule were low in DCs of nuclear factor RelB suppression,but significantly high comparing with mature DCs（P0.01）,and LPS RNAi RelB DC（that is,after RNAi RelB DC stimulated by LPS） surface of three class molecules also had a low level compared with LPS DC（P0.01）.To some extent,the expression levels of surface molecules in RNAi RelB DC were near to them of immature DC.T cells proliferation capacities and Th cytokines in DCs of nuclear transcription factor RelB suppression were significantly lower than those in mature DCs（P0.05）,having the same low levels with the immature DCs.It showed us that DCs of nuclear factor RelB suppression have the capacities which induce immunological tolerance in vitro.This is a novel pathway to research tolerogenic dendritic cells.

关 键 词：RELB 基因抑制 树突状细胞 免疫耐受 

分 类 号：R392.12[医药卫生—免疫学]