ICOS和PD-1协同刺激信号在四氯化碳致小鼠肝纤维化中的动态变化  

Dynamic changes of costimulatory signal ICOS and PD-1 in the progression of liver fibrosis in mice induced by carbon tetrachloride

作　　者：王波 李媛 萨如拉[2] 李慧 丁海涛 Wang Bo;Li Yuan;Sa Rula;Li Hui;Ding Haitao(Clinical Laboratory,Inner Mongolia People's Hospital,Hohhot 010017,China;School of Health Management,Inner Mongolia Medical University,Hohhot 010000,China;Inner Mongolia Center for Disease Control and Prevention,Hohhot 010030,China)

机构地区：[1]内蒙古自治区人民医院检验科,呼和浩特010017 [2]内蒙古医科大学公卫生管理学院,呼和浩特010000 [3]内蒙古自治区综合疾病预防控制中心,呼和浩特010030

出　　处：《国际免疫学杂志》2021年第3期245-251,共7页International Journal of Immunology

基　　金：内蒙古自治区自然科学基金(2020MS08155,2016BS0812);内蒙古自治区人民医院院内基金(2019YN13);内蒙古自治区人民医院博士科研启动资金(BS201801)。

摘　　要：目的探讨可诱导共刺激分子(inducible costimulatory molecular,ICOS)和程序性死亡受体1(programmed cell death protein 1,PD-1)蛋白在四氯化碳致小鼠肝纤维化中的动态变化。方法将80只雄性BALB/c小鼠按随机数字表法分为正常对照组和肝纤维化模型组,每组40只。流式细胞仪分析测定不同阶段滤泡辅助性T细胞(follicular helper cells,Tfh)群ICOS及PD-1的动态表达趋势及特征;酶联免疫吸附测定法(enzyme-linked immunosorbent assay,ELISA)检测小鼠不同周期脾淋巴细胞培养上清中的相关细胞因子白细胞介素(interleukin,IL)-21,IL-33,转化生长因子-β1(transforming growth factor-β,TGF-β1)的分泌水平及小鼠血清中Ⅲ型前胶原肽(procollagen typeⅢ,PCⅢ)的含量;分析PD-1^(+)CD4^(+)CXCR5^(+)T细胞、ICOS^(+)CD4^(+)CXCR5^(+)T细胞与肝纤维化标志物PCⅢ的相关性。结果与正常对照组相比,纤维化模型组中CXCR5^(+)CD4^(+)T细胞表面PD-1的表达比例均明显升高,并在13 W达到峰值,组间差异有统计学意义{(2.77±0.92)%比[8 W:(6.53±2.12)%,13 W:(27.47±5.15)%,20 W:(19.20±6.64)%],t值分别为4.60、13.35、6.93,P值均<0.05};CXCR5^(+)CD4^(+)T细胞表面ICOS的表达比例亦逐渐升高,13 W达到峰值,组间差异有统计学意义{(1.13±0.44)%比[8 W:(13.28±3.85)%,13 W:(33.62±6.28)%,20 W:(21.50±5.83)%],t值分别为8.87、14.60、9.85,P值均<0.05}。随着病程进展,肝纤维化小鼠组CD4^(+)CXCR5^(+)T细胞群IL-33,IL-21,TGF-β1的表达从8 W开始上调,13 W达到峰值,20 W仍维持较高水平,且均显著高于对照组,组间差异具有统计学意义(IL-33:t值分别为5.85、18.47和10.54,IL-21:t值分别为10.03、13.34和13.60,TGF-β1:t值分别为9.23、19.69和15.40,P值均<0.05)。PD-1^(+)CD4^(+)CXCR5^(+)T、ICOS^(+)CD4^(+)CXCR5^(+)T细胞表达水平与PCⅢ的水平成正相关(r值分别为0.6278和0.6036,P值均<0.05)。结论肝纤维化慢性期ICOS^(+)和PD-1^(+)的循环Tfh细胞可能促进肝纤维化发病,IL-21和IL-33作为�Objective To investigate the dynamic changes of inducible costimulator(ICOS)and programmed cell death protein 1(PD-1)in the progression of liver fibrosis in mice induced by carbon tetrachloride.Methods Total of 80 male BALB/c mice were randomly divided into normal control group and liver fibrosis model group with 40 mice in each group.The expression trends and characteristics of different periods of ICOS and PD-1 on the follicular helper cells(Tfh)were detected by flow cytometric analysis.The levels of interleukin(IL)-21,IL-33,transforming growth factor-β1(TGF-β1)in the supernatant of splenic lymphocytes from different cycles and the content of procollagen typeⅢ(PCⅢ)in serum were detected by enzyme linked immunosorbent assay(ELISA),and the correlation between the PD-1^(+)CD4^(+)CXCR5^(+)T cells,ICOS^(+)CD4^(+)CXCR5^(+)T cells and PCⅢwere also analyzed.Results Compared with the normal control group,the expression of PD-1 on the surface of CD4^(+)CXCR5^(+)T cells in the model group significantly increased,and reached the peak at 13 W{(2.77±0.92)%vs[8 W:(6.53±2.12)%,13 W:(27.47±5.15)%,20 W:(19.20±6.64)%],t values were 4.60,13.35 and 6.93 respectively,all P values<0.05};the expression of ICOS on the CXCR5^(+)CD4^(+)T cells also significantly increased,and reaching the peak at 13 W{(1.13±0.44)%vs [8 W:(13.28±3.85)%,13 W:(33.62±6.28)%,20 W:(21.50±5.83)%],t values were 8.87,14.60 and 9.85 respectively,all P values<0.05}.With the progression of the disease,the expression of IL-33,IL-21 and TGF-β1 on the surface of CD4^(+)CXCR5^(+)T cells in the model group increased from 8 W,and reached the peak at 13 W,and maintained a high level at 20 W,which were significantly higher than those in the control group(IL-33:t values were 5.85,18.47 and 10.54 respectively,IL-21:t values were 10.03,13.34 and 13.60 respectively,TGF-β1:t values were 9.23,19.69 and 15.40 respectively,all P values<0.05).The expression levels of PD-1^(+)CD4^(+)CXCR5^(+)T cells,ICOS^(+)CD4^(+)CXCR5^(+)T cells were positively correlated with the

关 键 词：肝纤维化 可诱导共刺激分子 程序性死亡受体1 白细胞介素-33 

分 类 号：R575.2[医药卫生—消化系统]