文章摘要
李丽,刘宝瑞,钱晓萍.钙离子载体和Poly(I:C)诱导人外周血来源的树突状细胞成熟的研究[J].南京医科大学学报,2007,(7):676~679
钙离子载体和Poly(I:C)诱导人外周血来源的树突状细胞成熟的研究
Generation of mature dendritic cells from peripheral blood mononuclear cells by Calcium ionophore and Poly(I:C) in vitro
投稿时间:2006-11-10  
DOI:10.7655
中文关键词: 树突状细胞  外周血单个核细胞  Poly(I:C)  钙离子载体A23187
英文关键词: dendritic cells  peripheral blood mononuclear cells  polyriboinosinic polyribocytidylic acid  calcium ionophore A23187
基金项目:南京市医学科技发展项目(ZKX0112)
作者单位
李丽 南京大学医学院附属鼓楼医院肿瘤中心,江苏 南京 210008 
刘宝瑞  
钱晓萍  
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中文摘要:
      目的:探讨从健康人外周血中体外诱导培养出成熟树突状细胞(dendritic cell, DC)的方法?方法:用密度梯度离心法分离人外周单个核细胞(PBMC),再用贴壁法获取单核细胞后加入GM-CSF和IL-4,培养5天后分5组:第1组为对照组,仅含有上述细胞因子;第2组,加入多聚次黄嘌呤胞嘧啶核苷酸[Poly(I:C)];第3组,加入钙离子载体(A23187);第4组,加入Poly (I:C)和CI;第5组,加入TNF-α?显微镜下观察培养细胞形态,流式细胞术检测DC表型,体外同种混合淋巴细胞反应(MLR)检测DC刺激T细胞的增殖活性?结果:第四组细胞具有典型的树突状细胞的特征,且高表达CD83?CD80?CD86和CD40分子,具有更强的刺激T细胞增殖能力?结论:经过组合细胞因子诱导能够培养出PBMC来源的DC,且经CI和Poly(I:C)进一步诱导后获得更成熟和功能更强DC?
英文摘要:
       Objective:To explore the generation of mature dendritic cells(DCs) induced from human peripheral blood monocytes in vitro. Methods:Monocytes were isolated from normal human peripheral blood mononuclear cells(PBMCs)and were separated into five groups after cultured with GM-CSF and IL-4 5 days later. The control group were added only GM-CSF and IL-4. One group were added polyriboinosinic polyribocytidylic acid[Poly(I:C)], one group were added calcium ionophore(CI),one group were added Poly(I:C) and CI, and the other group were added TNF-α. The morphology of cells was observed by microscope. The surface antigens of the induced cells were analyzed by fluorescence-activated cell sorting(FACS). T cell proliferating activity was determined by allo-MLR(mixed lymphocyte reaction) in vitro. Results:Large amount of mature DCs could be obtained from monocytes by culture in presence of GM-CSF,IL-4,Poly(I:C) and CI. DC expressed high level of CD83, CD80, CD86 and CD40, and were potent to stimulate the proliferation of allo-lymphocytes. Conclusion:The phenotypes and functions of DC generated from PBMC by the treatment of cytokine combination can be improved by calcium ionophore and Poly(I:C).
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