树突状细胞；体外培养；成熟度；激活性_

体外诱导生成树突状细胞的成熟度和激活性研究

http://www.100md.com 《河北北方学院学报》 2006年第2期

树突状细胞；体外培养；成熟度；激活性,,树突状细胞；体外培养；成熟度；激活性,1材料与方法,2结果,3讨论,参考文献

     【摘要】目的: 探讨人外周血单核细胞体外培养树突状细胞(dendritic cell,DC)的成熟度和激活性。方法: 从健康成人外周血分离单核细胞(PBM)，加入粒单系集落刺激因子(GMCSF)1000U/ml、重组白细胞介素4(IL4)500 U/ml，体外培养7d。然后加入肿瘤坏死因子a(TNFa)100ng/ml ，体外培养3d。培养流式细胞仪分别测定两次DC的主要组织相溶性复合体(MHC)Ⅱ类分子和协同刺激分子，分析其成熟度和激活度。结果:PBM经GMCSF和IL4诱导培养7d后，细胞成簇，表型为CD83 226% 、CD86 555% 、CD11c 361% 、CD64 32%、人类白细胞抗原(HLA)DR 134% ；加入TNFa培养3d后，细胞表型为CD83 815% 、CD86 993% 、CD11c 988% 、CD64 34%、人类白细胞抗原(HLA)DR 833% 。结论: GMCSF和IL4诱导培养PBM 7d，可获得大量不成熟DC，该体系有利于DC扩增，加入TNFa继续培养3d后，DC成熟度高，激活性好。

    【关键词】树突状细胞；体外培养；成熟度；激活性

    Dendritic Cells Maturity and Activation Induced in Vitro

    LIU XiaoXia, JIA LiPin, QIAO JunHong

    Department of Immunity and Pathogeny, Medical College,

    Hebei Engineering University, Handan 056029, Hebei China

    【ABSTRACT】 Objective: To study maturity and activation of dendritic cells (DC) induced from human peripheral blood monocytes(PBM). Methods: The monocytes collected from nomal peripheral blood were cultured with GMCSF (1000U/ml) and IL4 (500U/ml) for 7 days, then cultured with TNFα (100ng/ml) for 3 days. MHCⅡand costimulatory molecules of DC were analyzed by flow cytometry for two times, so maturity and activation of DC were known by them. Results: CD83、CD86、CD11c、CD64、HLADR expressions in PBM were 226%、555%、361%、32%、134% after PBM were cultured with GM-CSF and IL4 for 7 days, but their expressions in PBM were 815%、993%、988%、34%、833% after PBM were cultured with TNFαfor 3 days. Conclusion: DC induced from PBM with GMCSF and IL4 for 7days were not mature, but the system inducement was beneficial to expansion of DC. Cultured with TNFαfor 3 days , DC induced form PBM were very good in maturity and activation. ......

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