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作 者:郭根凯[1] 梁军[1] 侯亚义[2] 李慧[1] 周康新[1] 孙凌云[1]
机构地区:[1]南京大学医学院附属鼓楼医院风湿免疫科,210008 [2]南京大学医学院免疫学教研室
出 处:《中华风湿病学杂志》2007年第9期524-527,共4页Chinese Journal of Rheumatology
基 金:教育部高等学校博士点基金资助项目(20050315001);江苏省135重点人才基金资助项目(RC2002003);江苏省自然科学基金资助项目(BK2006007)
摘 要:目的:探讨系统性红斑狼疮(SLE)患者骨髓间质干细胞(MSCs)体外对B淋巴细胞的免疫调节作用是否存在异常。方法:采用直接贴壁法分离培养正常人及SLE患者MSCs。免疫磁珠法分选正常人外周血B淋巴细胞,与MSCs共培养,流式细胞术检测B细胞凋亡及表面标志表达,3H掺入法检测B细胞增殖,酶联免疫吸附法(ELISA)检测培养上清中免疫球蛋白IgG、IgM和IgA水平。结果:①B细胞+正常MSCs共培养组B细胞增殖(4307±308)cpm较单纯B细胞培养组(7059±346)cpm降低(P〈0.05);②B细胞+正常MSCs共培养组B细胞凋亡率(14±7)%低于单纯B细胞培养组(25±8)%(P〈0.05);③B细胞+正常MSCs共培养组B细胞表面CD86表达率(53±5)%较单纯B细胞培养组(66±10)%低(P〈0.05);④与MSCs共培养后,B细胞分泌免疫球蛋白IgG、IgM和IgA较单纯B细胞培养显著减少(P〈0.05);⑤B细胞+狼疮MSCs共培养组与B细胞+正常MSCs共培养组比较,B细胞增殖、凋亡、CD86表达和Ig分泌两组间差异无统计学意义。结论:MSCs在体外对B细胞有抑制作用,SLE患者骨髓MSCs与正常MSCs相比,对B细胞的表面刺激分子表达、增殖、Ig分泌有相似的抑制作用。Objective To investigate the immunoregulatory effects of bone marrow derived mesenchymal stem cells (MSCs) on B lymphocytes in systemic lupus erythematosus (SLE). Methods Human bone marrow MSCs were isolated from SLE patients and normal controls and cultured ex vivo. B lymphocytes were purified from normal peripheral blood mononuclear cells by using CD19 microbeads. Purified B cells were cocultured in 96-well flat-bottom plates with MSCs. B cell apoptosis and surface markers were analyzed by flow cytometry. B cell proliferation was assessed by [^3H]-thymidine incorporation. ELISA was used to measure supernatant levels of IgG, IgM and IgA. Results ①B cell proliferation in normal MSCs coculture group (4307± 308) cpm was significantly inhibited compared to simple B cell group (7059±346) cpm (P〈0.05);②B cell apoptotic percentage in normal MSCs cocuhure group (14±7)% was lower than in B cell group (25±8)% (P〈 0.05); ③The percentage of CD86 molecule expression on B cells in normal MSCs coculture group (53±5)% was lower than that in simple B cell group (66±10)% (P〈0.05);④The levels of IgG, IgM and IgA secreted by B cell decreased when cocuhured with MSCs (P〈0.05); ⑤There were no significant differences of B cell proliferation, apoptosis, CD86 expression and immunoglobulin production regulated by MSCs from lupus patients and normal controls. Conclusion MSCs show an inhibitory effect on B cells. Both MSCs from SLE patients and normal MSCs have similar patterns in B cell proliferation, activation and immunoglobulin production.
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