【作者】 陈伟；

【导师】 李向培；

【作者基本信息】 安徽医科大学 ， 内科学， 2007， 硕士

【摘要】 目的检测系统性红斑狼疮(SLE)患者外周血CD4+CD25+调节性T细胞(Treg)和CD4+CD25highT细胞占CD4+T细胞的比率以及血清细胞因子IL-4、IFN-γ、IL-10、IL-12及TGF-β的浓度变化,探讨它们在SLE发病机制中的作用和临床意义。方法选择SLE患者54例,均符合1997年ACR修订SLE分类标准,其中未治疗的初发SLE组31例,激素治疗SLE组为23例,选择20例健康志愿者作对照。用酶联免疫吸附试验ELISA方法检测SLE患者以及正常对照的细胞因子IL-4、IFN-γ、IL-10、IL-12及TGF-β的血清浓度;采用流式细胞仪检测54例SLE患者及20例正常对照外周血CD4+CD25+T细胞和CD4+CD25highT细胞比率,比较SLE患者不同组别以及SLE不同脏器损害时的Treg比率及细胞因子浓度变化的差异。结果1.SLE患者CD4+CD25highT细胞的比率低于正常对照组(P<0.05),CD4+CD25+T细胞的比率与正常对照组比较无明显差异(P>0.05)。SLE患者血清IL-4、IFN-γ、IL-10、IL-12的表达水平及IL-4/IFN-γ的比值均高于正常对照组(P<0.01),血清TGF-β的表达水平显著低于正常对照组(P<0.01)。2.初发SLE组CD4+CD25+T细胞及CD4+CD25highT细胞的比率均低于正常对照(P<0.05,P<0.01);初发SLE组与治疗SLE组之间比较其比率均无明显差异(P>0.05);抗ds-DNA(+)组CD4+CD25highT细胞的比率低于正常对照(P<0.05)。初发SLE组IL-4、IFN-γ、IL-10、IL-12的表达水平均明显高于正常对照(P<0.01),治疗SLE组IL-4、IL-10、IL-12的表达水平均明显高于正常对照(P<0.01),IFN-γ的表达水平高于正常对照(P<0.05),初发及治疗SLE组血清TGF-β的表达水平均显著低于正常对照组(P<0.01);初发与治疗SLE组比较,初发SLE组IFN-γ、IL-10的表达水平均高于治疗组(P<0.05);初发SLE组的IL-4、IL-12及TGF-β表达水平与治疗组相比均无明显差异(P>0.05)。3.双变量相关分析显示SLE组CD4+CD25highT细胞与血清IL-12的表达水平呈负相关(r=-0.195,P<0.05);初发SLE组CD4+CD25highT细胞与血清IL-10、IL-12的表达水平均呈负相关(r=-0.238,P<0.05;r=-0.242,P<0.05)。偏相关分析发现以IL-4作为控制变量时,CD4+CD25+T细胞与IL-4/IFN-γ比值正相关(r=0.241,P=0.040);根据实验室指标进行相关分析发现,SLE患者CD4+CD25+T细胞与抗C1q抗体呈负相关(r=-0.323,P=0.031)。结论SLE患者外周血中CD4+CD25+T细胞和CD4+CD25highT细胞的比率减少,它们可能参与了SLE的发病。血清IL-4、IFN-γ、IL-10、IL-12表达水平及IL-4/IFN-γ比值增高,TGF-β的表达水平降低, Th1、Th2及Th3型细胞因子均可能参与了SLE疾病的发生发展。Treg与IL-10、IL-12及IL-4/IFN-γ比值存在不同程度的相关性,提示Treg在引起SLE免疫异常的作用中可能对Th1/Th2的平衡有一定的影响,依赖于某些相关细胞因子,从而参与了SLE的发病机制。临床上联合检测SLE患者外周血中CD4+CD25+T细胞和CD4+CD25highT细胞的比率及血清IL-4、IFN-γ、IL-10、IL-12、TGF-β的表达水平,对了解SLE的免疫异常、病情的评估及指导治疗具有重要的临床意义。更多还原

【Abstract】 Objective To investigate the proportions of CD4+CD25+ regulatory T cells and CD4+CD25highT cells in peripheral blood CD4+T cells, and the levels of cytokines (IL-4,IFN-γ,IL-10,IL-12 and TGF-β)in systemic lupus erythematosus(SLE), then to explore the role of regulatory T cells in the pathogenesis of SLE.Methods Fifty four SLE patients (including thirty one Recent-onset SLE patients and twenty three treated SLE patients) and twenty normal controls were selected. Clinical data and blood were collected and sandwich ELISA was employed for cytokines of IL-4,IFN-γ,IL-10,IL-12 and TGF-βin blood serum. Flow cytometric analysis was employed for the proportions of CD4+CD25+ regulatory T cells and CD4+CD25highT cells in CD4+T cells in the peripheral blood of SLE(n=54) and healthy volunteers(n=20).The interaction and association between the proportions of peripheral blood regulatory T cells and the levels of cytokines,with SLE organ damage were analyzed.Results 1.The proportions of CD4+CD25highT cells in SLE was lower than that in healthy controls(P<0.05).There was no significant difference between CD4+CD25+T cells and that in controls(P>0.05).The levels of IL-4, IFN-γ, IL-10,IL-12 and the IL-4/IFN-γration in SLE group were higher than those in healthy controls(P<0.01).The levels of TGF-βration in SLE group was lower than those in healthy controls(P<0.01). 2.The proportions of CD4+CD25+T cells and CD4+CD25highT cells in recent-onset SLE were significantly lower than those in healthy controls(P<0.05,P<0.01).There was no significant difference between CD4+CD25+T cells and that in treated patients(P>0.05).The proportions of CD4+CD25highT cells in patients with anti ds-DNA(+) antibody SLE group was lower than healthy controls(P<0.05). The levels of IL-4,IFN-γ,IL-10 and IL-12 in recent-onset SLE were higher than those in healthy controls(P<0.01). The levels of IL-4,IL-10,IL-12 in treated SLE were higher than those in healthy controls(P<0.01). The level of TGF-βin recent-onset SLE and treated SLE were significantly lower than that from normal controls(P<0.01). The levels of IFN-γand IL-10 in recent-onset SLE was higher than those in treated SLE(P<0.05). There was no significant difference of IL-4,IL-12 and TGF-βbetween recent-onset SLE and that in treated SLE (P>0.05). 3.Bivariate correlation analysis indicated that CD4+CD25highT cells is negatively correlated with IL-12(r=-0.195,P<0.05).The proportions of CD4+CD25highT cells in recent-onset SLE is negtively correlated with IL-10 and IL-12(r=-0.238,P<0.05;r=-0.242,P<0.05).When IL-4 was taken as the controls variable, CD4+CD25+ regulatory T cells was positively correlated with the IL-4/IFN-γratio(r=0.241,P=0.040). The proportions of CD4+CD25+T cells in SLE is negtively correlated with anti-C1q(r=-0.323,P=0.031).Conclusion The proportions of CD4+CD25+T cells and CD4+CD25highT cells are decreased in serum of SLE,The levels of IL-4,IFN-γ,IL-10,IL-12 and IL-4/IFN-γratio are increased while lower TGF-βthan those in normal controls in SLE. The Th1、Th2 and Th3 might participate in the pathogenesis of SLE.The regulatory T cells possibly play a role in the pathogenesis of SLE and in the Th1/Th2 balance.It has great importance to combined detection of IL-4,IFN-γ,IL-10,IL-12,TGF-βand the proportions of CD4+CD25+T cells and CD4+CD25highT cells in CD4+T cells in the peripheral blood of SLE for evaluation the immunity disturbance,the damage of organs and the disease activity.更多还原