IL-27诱导原发性胆汁性肝硬化患者CD4^＋T细胞增殖分化作用机制研究  

Proliferation and differentiation of CD4^＋ T cells induced by IL-27 in primary biliary cirrhosis

作　　者：蒋廷旺 韩志君[2] 熊怀民 盛建华 张红星 龚燕萍 仲人前[3] 

机构地区：[1]常熟市医学检验所,215500 [2]无锡市第二人民医院检验科,214002 [3]上海长征医院实验诊断科全军临床免疫中心

出　　处：《中华微生物学和免疫学杂志》2011年第11期1023-1026,共4页Chinese Journal of Microbiology and Immunology

基　　金：苏州市科技发展计划应用基础研究项目（SYSD2010027）;常熟市科技发展计划（CR201113）

摘　　要：目的 探讨IL-27对原发性胆汁性肝硬化（primary biliary cirrhosis,PBC）患者外周血CD4＋T细胞的增殖分化作用及其相关免疫学机制.方法 收集PBC患者、慢性乙肝患者（choronic hepatitis B,CHB）、健康体检者（health controls,HCs）外周血,磁珠分离CD4＋T细胞.IL-27体外作用后,CCK-8测定细胞增殖情况,ELISA法检测细胞因子,定量PCR分析T-bet和GATA3基因表达情况,免疫印迹测定p-STAT-1和p-STAT-3的表达.结果 IL-27作用后,PBC组、CHB组和HCs组CD4＋T细胞增殖能力均显著增强,PBC组CD4＋T细胞增殖能力强于CHB和HCs组,差异有统计学意义（P＜0.001）,同时PBC组细胞培养液中IL-2和IFN-γ在IL-27作用后较CHB和正常对照组均显著增高（P＜0.001）,IL-10表达无明显变化.未经IL-27诱导情况下,PBC组T-bet表达高于CHB组（P=0.007）,IL-27诱导后PBC组CD4＋T淋巴细胞T-bet基因表达显著增加,同时对GATA3具有抑制作用,作用前后差异有统计学意义（P＜0.001）,CHB组作用前后无显著变化（P=0.3）.免疫印迹测定p-STAT-1、p-STAT-3发现,正常情况下各研究组均不表达p-STAT-1、p-STAT-3,IL-27作用后,表达明显升高,其中PBC组升高尤为明显.结论 IL-27可以诱导PBC患者CD4＋T细胞增殖,并通过活化p-STAT-1、p-STAT-3信号通路,诱导CD4＋T细胞向Th1分化,同时分泌相关细胞因子,可能在PBC早期免疫炎症反应过程中具有重要作用.Objective To investigate mechanisms for IL-27 induced proliferation and differentiation of peripheral blood CD4^＋ T cells in primary biliary cirrhosis （PBC）.Methods Periperal blood CD4^＋ T cells were isolated from patients with PBC,chonic hepatitis B （CHB） and health controls （HCs）.After IL-27 stimulation,proliferation ability of CD4^＋ T cells was evaluated by CCK-8 kit,and cytokines were analyzed by ELISA.Real-time PCR was employed to assay mRNA expression of T-bet and GATA3 in CD4^＋ T cells.p-STAT-1 and pSTAT-3 expression in CD4^＋ T cells were detected by Western blot.Results Enhanced proliferation of CD4^＋ T cells was found in all subjects after IL-27 stimulation.However,the proliferation ability in patients with PBC was greater than that in CHB and HCs （ P〈0.001 ）.Levels of IL-2 and IFN-γ in supernatant from IL-27-incubated PBC blood CD4^＋ T cells were higher than that from CHB and HCs （P〈0.001 ）.In normal situation,T-bet mRNA of CD4^＋ T cells in PBC group was higher than that in CHB group （P=0.007）.Furthermore,after IL-27 stimulation,elevated T-bet mRNA expression and GATA3 inhibition were found in patients with PBC.High expression of p-STAT-1 and p-STAT-3 in blood CD4^＋ T cells were found in PBC,CHB and HCs after stimulation by IL-27.But their expression in patients with PBC were higher than those in patients with CHB and HCs.Conclusion Proliferation of blood CD4^＋ T cells could be induced by IL-27 in patients with PBC.The signaling pathways of p-STAT-1,p-STAT-3 were involved to induce Th1 immune response and related cytokines expression.This study implicated that IL-27 may play important roles in early inflammation damage in PBC.

关 键 词：白细胞介素-27 原发性胆汁性肝硬化 CD4^+ T淋巴细胞 

分 类 号：R575.2[医药卫生—消化系统]