RelB基因沉默对骨髓树突细胞生物学效应的影响  

Biological effects of RelB gene silencing on the dendritic cells derived from bone marrow

作　　者：郑磊[1] 颜晓慧[2] 王前[1] 包杰[1] 平宝红[3] 

机构地区：[1]南方医院检验医学中心,广州510515 [2]南方医院临床研究中心,广州510515 [3]南方医院惠侨科,广州510515

出　　处：《解放军医学杂志》2008年第3期336-338,共3页Medical Journal of Chinese People's Liberation Army

基　　金：广东省自然科学基金资助课题(04020404)

摘　　要：目的探讨核转录因子NFκ-B/RelB基因沉默对树突细胞(DC)生物学效应的影响。方法采用细胞因子诱导培养法体外扩增小鼠骨髓DC,并经免疫磁珠纯化,获得高纯度骨髓DC后,行RelB基因沉默,获得RNAi RelB-DC,扫描电镜和透射电镜观察细胞形态特点,并利用流式细胞术分析细胞表面分子的表达水平,同种异基因混合淋巴细胞反应(MLR)法分析RNAi RelB-DC刺激异基因T细胞增殖的能力。结果扫描电镜下RNAi RelB-DC细胞表面突起较少而短,细胞表面多皱褶;透射电镜下可见细胞内多吞噬泡样结构;流式细胞术分析显示细胞表面分子MHC-II、CD86和CD40的表达水平相对较低;MLR显示该DC刺激异基因T细胞增殖的能力相对较弱。结论RelB基因沉默DC的生物学效应表现为未成熟DC的特点,这种RNAi RelB-DC作为一种新型的致耐受DC,有望应用于免疫耐受的诱导研究。Objective To study the biological effects of nucleic transcript factor NF-κB/RelB gene silencing on bone marrow derived dendritic cells （BMDCs）. Methods For constructing a new type of DCs, RNAi RelB DC, raouse BMDCs were cultured through IL-4 and GM-CSF cytokine inducing system and were purified with CD11c positive immunomicrobeads by flow cytometer, and RelB gene was silenced with PCR expression cassettes. The RelB RNA interference effect in DCs was proved at both mRNA and protein levels. The biological characteristics of RNAi RelB DCs were investigated by means of cell morphology, cell surface markers and mixed lymphocyte reaction ability. The cell microstructure was observed using scanning electron microscopy and transmission electron microscopy. The cell surface markers, such as MHC-Ⅱ, CD86 and CD40, were detected by flow cytometer. The mixed lymphocyte reaction ability, as well as the ability of DCs in stimulating allogeneic T cells proliferation, was detected by ^3H-TdR incorporation method. Results RNAi RelB DCs seemed to have fewer and shorter spikes and many folds under scanning electron microscopy. Many phagocytic vacuoles were observed in the cytoplasm under transmission electron microscopy. DCs surface molecules, MHC-Ⅱ, CD86 and CD40, were expressed at a relatively lower level compared with the control DCs. This RNAi RelB DCs also showed weak allogeneic T cells stimulating ability in vitro in MLR. Condusion The dendritic cells derived from bone marrow are characterized the biological features and immunological function of immature DC with silencing of RelB gene. This kind of RNAi RelB DCs may be used to induce immunotolerance as a novel tolerogenic DCs.

关 键 词：RELB 基因沉默 树突细胞 

分 类 号：R329.25[医药卫生—人体解剖和组织胚胎学]