SLE患者CD4^+CD25^-T细胞来源的体外诱导型Treg细胞IL-10表达特征的初步研究

Preliminary study about the expression of IL-10 on induced Treg cells from CD4^+CD25^-T cells of SLE patients in vitro

目的 :探讨白细胞介素(interleukin,IL-10)在系统性红斑狼疮(systemic lupus ergthematosu,SLE)患者体外诱导型调节性T细胞(induced regulatory T cell,i Treg)中表达的变化。方法:分离健康对照和SLE患者外周血单个核细胞,并进一步分离出CD4^+CD25^-T细胞,将其在体外以TGF-β诱导转化;用流式细胞术分析诱导转化后T细胞中CD25^+和CD25^-、Foxp3^+和Foxp3-细胞亚群胞内IL-10(introcellular IL-10,iIL-10)和细胞膜表面IL-10(membrane IL-10,m IL-10)的表达。结果:(1)SLE患者和正常人经诱导转化的CD4^+亚群细胞内CD25^+T细胞表达iIL-10均明显增高,且患者显著高于正常人;(2)同样,患者和正常人诱导后CD4^+亚群细胞内CD25^+T细胞表达m IL-10均明显增高,但患者与正常人表达水平相似;(3)SLE患者和正常人经诱导转化形成的CD4^+CD25^+亚群细胞内Foxp3^+T细胞表达iIL-10亦均明显增高,且患者显著高于正常人;(4)SLE患者和正常人经诱导转化形成的CD4^+CD25^+亚群细胞内Foxp3^+T细胞表达m IL-10也明显增高,但患者和正常人类似;(5)SLE患者以上各项改变均与SLE疾病活动性指数无相关性。结论:正常人经诱导转化形成的CD4^+CD25^+Foxp3^+Treg细胞与IL-10表达密切相关。SLE患者诱导转化后形成的i Treg细胞以及未转化的CD4^+CD25^-T细胞内表达iIL-10的水平均显著高于正常人;但是细胞膜上的m IL-10表达与正常人没有差异。

Objective： To investigate the level of IL-10 expressing on induced Treg cells （iTreg） such as CD4＋CD25＋T cells of SLE patients.Methods： Peripheral blood mononuclear cells （PBMCs） were isolated from healthy donors and SLE patients,respectively,and CD4＋CD25-T cells were further isolated.Then the cells were induced and transformed by TGF-β in vitro; expression of intracellular and membrane IL-10 on CD25＋ and CD25-,Foxp3＋ and Foxp3-subsets of induced and transformed T cells were analyzed by flow cytometry.Results： Expression level of the intracellular IL-10 （iIL-10） in CD25＋ T cell subsets was obviously increased in both of the induced and transformed CD4＋T cells from the normal controls and SLE patients,but it was significantly higher in the SLE patients.In the induced and transformed CD4＋T cells both from the normal controls and SLE patients,expession level of the membrane IL-10 in CD25＋T cell subsets was obviously increased,while it had no difference between the SLE patients and the control group.In the normal subjects and SLE patients,the expession level of intracellular IL-10 in the induced and transformed CD4 ＋CD25 ＋ Foxp3＋T cells subsets was obviously increased,and was significantly higher in the SLE patients than that of the control group.The expession level of membrane IL-10 was increased in the induced and transformed CD4＋CD25＋ Foxp3＋T cells subsets,while no difference was observed between normal subjects and SLE patients.All the changes mentioned before have no correlation with SLEDAI.Conclusion： The IL-10 expression level was closely correlated with the induced and transformed CD4 ＋CD25＋ Foxp3＋ Treg cells from the normal subjects.Abnormal expression of intracellular IL-10 was observed on the induced and transformed Treg cells from the CD4 ＋CD25-T cell of SLE patients.The intracellular IL-10 exprssion level of CD4＋CD25＋Foxp3＋ Treg cells from the SLE patients was significantly higher than that of the control group.And the intracellular IL-10 exprssion level of CD4＋CD25＋ Foxp3-Treg cells and CD4＋CD25-T cell without induction was also significantly higher than that of the control group; But the expessions level of membrane IL-10 was not different between the SLE patients and the normal controls.