Allo-PBSCT患者CD4^+CD25^+调节性T细胞的体外研究  

Study on post-allogeneic peripheral blood stem cell transplantation patients' CD4^+CD25^+ regulatory T cells in vitro

作　　者：翟海龙[1] 赖永榕[2] 

机构地区：[1]黄石理工学院医学院,湖北黄石435003 [2]广西医科大学第一附属医院

出　　处：《山东医药》2011年第33期10-12,共3页Shandong Medical Journal

摘　　要：目的探讨异基因外周血干细胞移植(Allo-PBSCT)患者外周血CD4+CD25+调节性T细胞(Tregs)在协同刺激信号作用下的增殖反应、与CD4+CD25-T细胞混合淋巴细胞反应及上述两种培养细胞的细胞因子分泌情况。方法对36例Allo-PBSCT患者离体CD4+CD25+Tregs在抗CD3-mAbs和抗CD28-mAbs的刺激下行CD4+CD25+Tregs培养和CD4+CD25+Tregs、CD4+CD25-T细胞混合淋巴细胞培养72 h,然后加入CCK-8溶液孵育1 h,用酶标仪检测OD450值,同时测量上述两种细胞培养上清液中IL-10、TGF-β和IFN-γ的浓度。另以7例健康者为对照组。结果 CD4+CD25+Tregs细胞OD450值低于CD4+CD25-T细胞(P<0.01)。CD4+CD25+Tregs与CD4+CD25-T细胞混合OD450值也低于CD4+CD25-T细胞(P<0.01)。有移植物抗宿主病(GVHD)和无GVHD患者离体CD4+CD25+Tregs在抗CD3-mAbs和抗CD28-mAbs的共刺激下分泌的IL-10、TGF-β和IFN-γ低于CD4+CD25-T细胞(P<0.01),有GVHD和无GVHD患者的两细胞混合组分泌的这3种细胞因子均低于CD4+CD25-T细胞(P<0.05),Allo-PBSCT患者分泌细胞因子情况与对照组类似。结论有效地利用Allo-PBSCT受者外周血CD4+CD25+Tregs的抑制功能,则有可能降低Allo-PBSCT后GVHD的发病率。Objective To investigate the proliferation reaction of CD4＋CD25＋ Tregs in the stimulating of costimulatory signal,lymphocyte reactions mixed with CD4＋CD25-T cells of CD4＋CD25＋ Tregs,and cytokine secretion state of the two cells in allogeneic peripheral blood stem cell transplantation（Allo-PBSCT） patients.Methods CD4＋CD25＋Tregs and CD4＋CD25-T cells from peripheral blood obtained from 36 patients who had undergone Allogeneic peripheral blood stem cell transplantation（Allo-PBSCT）,7 healthy volunteers as control,were isolated with magnetic cells sorting separation.Then CD4＋CD25＋Tregs and CD4＋CD25＋Tregs＋CD4＋CD25-T cells were cultered for 72 hours,stimulated by anti-CD3-mAbs and anti-CD28-mAbs.After that the cultures added with CCK-8 solution were incubated for 1 hour.Then OD450 were detected by ELISA.IL-10,TGF-β and IFN-γ from the two above cell cultures were detected by ELISA method.Results OD450 values of CD4＋CD25＋Tregs were both extremely lower than that of CD4＋CD25-T cells and CD4＋CD25＋Tregs＋CD4＋CD25-T cells（P〈0.01）.IL-10,TGF-β and IFN-γ secreted by CD4＋CD25＋Tregs in vitro from patients with and without GVHD were signigicantly lower than that of CD4＋CD25-T cells（P〈0.01）.The 3 cytokines secreted by CD4＋CD25＋Tregs＋CD4＋CD25-T cells group were also signigicantly lower than that of CD4＋CD25-T cells（P〈0.05）.The cytokines secretory of Allo-PBSCT group was similar with that of control group.Conclusions If the suppressive function of CD4＋CD25＋Tregs are utilized,incidence of GVHD post-Allo-PBSCT may decrease.

关 键 词：干细胞移植 异基因 移植物抗宿主病 CD4+CD25+调节性T细胞 增殖 体外 

分 类 号：R733.7[医药卫生—肿瘤]