间充质干细胞对狼疮肾炎T淋巴细胞的免疫调节作用  

Immunoregulatory effects of mesenchymal stem cells on T-lymphocytes of lupus nephritis

作　　者：裘影影[1] 李晶[1] 何建强[2] 殷玉俊[1] 汤郁[1] 尤海燕[1] 

机构地区：[1]江苏大学附属医院风湿科,镇江212001 [2]江苏大学附属医院肾内科

出　　处：《中华风湿病学杂志》2009年第6期372-375,I0001,共5页Chinese Journal of Rheumatology

基　　金：江苏省科技厅社会发展项目（BS2005043）;江苏省镇江市重点医学人才资助项目（2005009）;江苏省镇江市科技局社会发展项目（SH2004040）

摘　　要：目的探讨间充质干细胞（MSCs）在体外对狼疮肾炎（LN）外周血T淋巴细胞的免疫调节作用。方法从人骨髓中分离培养MSCs，采用流式细胞仪（FCM）分析鉴定MSCs的纯度。在植物血凝素（PHA）刺激下，LN外周血T淋巴细胞与不同数量的MSCs共培养。分组：A组：T淋巴细胞；B组：MSCs1＋T淋巴细胞（MSCs1：T=1：5）；C组：MSCs2＋T淋巴细胞（MSCs2：T=1：20）；D组：MSCs3＋T淋巴细胞（MSCs3：T=1：100）。用四甲基偶氮唑蓝（MTT）比色法检测各组T淋巴细胞的增殖情况，FCM分析各组T淋巴细胞CD28和CD152的表达及T淋巴细胞凋亡情况，实时定量反转录-聚合酶链反应（RT—PCR）检测各组T淋巴细胞的干扰素（IFN）-γ、白细胞介素（IL）-10、转化生长因子（TGF）-β1基因的水平。结果在体外，MSCs对由PHA诱导的LNT淋巴细胞的增殖、凋亡均有抑制作用，且抑制作用与MSCs呈剂量依赖性。MSCs抑制T淋巴细胞CD28表达，对CD152的表达无明显影响。MSCs能促进LN患者T淋巴细胞TGF—β1基因的表达，抑制IL-10、IFN-γ基因的表达。结论MSCs可能通过抑制T淋巴细胞增殖、减少T淋巴细胞凋亡、抑制T淋巴细胞CD28表达和促进T淋巴细胞TGF—β，基因表达及抑制IL-10、IFN-1基因表达来下调LN的免疫反应。Objective To investigate the immunoregulatory effects of mesenchymal stem ceils （MSCs） on the peripheral T-lymphocytes of lupus nephritis in vitro. Methods MSCs were isolated and expanded from human bone marrow cells. The purity of MSCs was identified by flow cytometry （FCM）. The MSCs （4×10^4, 1× 10^4, 2×10^3） were added into wells containing peripheral blood lymphocytes （2×10^5） from lupus nephritis in the presence of phytohemagglutinin [PHA）. Samples were divided into the following groups： group A：T- lymphoeytes alone; group B： MSCsl with T-lymphocytes（MSCsl：T =1：5）; group C： MSCs2 with T-lymphocytes （MSCs2 ：T =1：20）; group D： MSCs3 with T-lymphocytes （MSCs3 ：T =1：100）. The proliferation of T- lymphocytes was assessed by MTT. FCM was used to analyze the apoptosis of T-lymphocytes and surface markers of CD28 and CD152. The gene expression of interferon -γ （IFN-γ）, interleukin-10 （IL-10）, transforming growth factor-β1 （TGF-β1） were detected by real-time RT-PCR. Results In vitro, with the presence of MSCs, peripheral blood T-lymphocytes from lupus nephritis were statistically significantly decr- eased in their proliferative activities , apoptosis and CD28 expression in a dose-dependent manner. No inhibitmy effects on CD152 expression of T-lymphocytes had been observed . MSCs promoted the gene expression of TGF-β 1 and inhibited the gene expression of IL-10, IFN-γ. Conclusion MSCs can inhibit the proliferative activities, apoptosis and CD28 expression of peripheral blood T-lymphocytes of lupus nephritis, increase gene expression of TGF-β and lower the gene expression of IL-10, IFN-γ which may play an important role in it＇s immunosuppressive effects on lupus nephritis.

关 键 词：狼疮肾炎 间质干细胞 T淋巴细胞 

分 类 号：R686[医药卫生—骨科学]