CX3CL1/CCL26-CX3CR1通路在原发性胆汁性胆管炎免疫机制中的作用  

Role of CX3CL1/CCL26-CX3CR1 pathway in the immunological mechanism of primary biliary cholangitis

作　　者：曹逸涵 孙晓川 陈志磊 张硕 赵丽伶 孙金磊 邵体红 董振华[2] 陈华 邓垂文 常晓燕[3] 李永哲[4] 杨云娇 费允云 王立 张奉春 CAO Yi-han;SUN Xiao-chuan;CHEN Zhi-lei;ZHANG Shuo;ZHAO Li-ling;SUN Jin-lei;SHAO Ti-hong;DONG Zhen-hua;CHEN Hua;DENG Chui-wen;CHANG Xiao-yan;LI Yong-zhe;YANG Yun-jiao;FEI Yun-yun;WANG Li;ZHANG Feng-chun(Department of Rheumatology,Key Laboratory of Rheumatology and Clinical Immunology,Ministry of Education,National Clinical Research Center for Dermatologic and Immunologic Diseases(NCRC-DID),Peking Union Medical College Hospital,Chinese Academy of Medical Sciences&Peking Union Medical College,Beijing 100730,China;Department of Traditional Chinese Medicine,Peking Union Medical College Hospital,Chinese Academy of Medical Sciences&Peking Union Medical College,Beijing 100730,China;Department of Pathology,Peking Union Medical College Hospital,Chinese Academy of Medical Sciences&Peking Union Medical College,Beijing 100730,China;Department of Clinical Laboratory,Peking Union Medical College Hospital,Chinese Academy of Medical Sciences&Peking Union Medical College,Beijing 100730,China)

机构地区：[1]中国医学科学院,北京协和医学院,北京协和医院风湿免疫科,风湿免疫病学教育部重点实验室,国家皮肤与免疫疾病临床医学研究中心,北京100730 [2]中国医学科学院,北京协和医学院,北京协和医院中医科,北京100730 [3]中国医学科学院,北京协和医学院,北京协和医院病理科,北京100730 [4]中国医学科学院,北京协和医学院,北京协和医院检验科,北京100730

出　　处：《中华临床免疫和变态反应杂志》2020年第3期183-189,共7页Chinese Journal of Allergy & Clinical Immunology

基　　金：中国医学科学院中央级公益性科研院所基本科研业务费专项资金资助(2019XK320022);国家自然科学基金青年项目(81501414,81501410);首发科研专项青年项目(2016-4-40111);国家重点研发计划:干细胞移植治疗典型免疫疾病的创新策略及应用实践(2016YFA010100);大学生创新训练计划项目(2017zlgc0604)。

摘　　要：目的探究CX3CL1/CCL26-CX3CR1通路在原发性胆汁性胆管炎(primary biliary cholangitis,PBC)免疫机制中的作用。方法收集PBC患者和健康对照者外周血及肝脏病理标本,采用ELISA检测血浆CX3CL1及CCL26水平,流式细胞术检测外周血单个核细胞各亚群中CX3CR1+细胞比例。使用免疫组化分析肝活检组织中CX3CL1和CCL26表达情况。采用ELISA和流式细胞术检测不同细胞因子及LPS刺激下,体外培养的人肝内胆管上皮细胞中CX3CL1/CCL26-CX3CR1通路表达水平变化。结果共纳入PBC患者40例、健康对照者18例。PBC患者血浆CX3CL1水平[(0.690±0.271)ng/mL]较健康对照者[(0.540±0.101)ng/ml]显著升高(P=0.044)。PBC患者血浆CCL26浓度[(8.94±4.09)pg/mL]较健康对照者[(6.75±1.86)pg/mL]有升高趋势,但无统计学差异(P=0.104)。与健康对照者相比,PBC患者CX3CR1表达水平在NKT-like细胞[(63.5±25.4)%vs.(78.6±18.0)%,P=0.026]和CD4^+T细胞[(5.5±5.4)%vs.(13.7±9.0)%,P=0.003]中显著增高,且后者这一差异在其CD28+和CD28-亚群中均显著。PBC患者肝脏胆管上皮细胞同时高表达CX3CL1和CCL26,健康对照者则弱表达CX3CL1,无CCL26表达。人肝内胆管上皮细胞CX3CL1表达水平在IFN-γ刺激下显著增加,CCL26表达在IL-4、IL-13刺激下显著增加;在IFN-γ刺激下,CX3CR1表达显著升高。结论PBC患者肝内胆管上皮细胞可能在IFN-γ刺激下CX3CL1表达增加,在IL-4和IL-13刺激下CCL26表达增加,其受体CX3CR1在外周血NKT-like细胞和CD4^+T细胞中表达上调。CX3CL1和CCL26可能通过CX3CR1协同介导PBC胆管上皮损伤。Objective To investigate the role of CX3CL1/CCL26-CX3CR1 pathway in the immunologi-cal mechanism of primary biliary cholangitis(PBC).Methods Peripheral blood and liver tissue samples from patients with PBC and healthy controls(HCs)were collected.The plasma levels of CX3CL1 and CCL26 were determined by ELISA.Flow cytometry was conducted to measure the percentages of CX3CR1+cells in different subsets of peripheral blood mononuclear cells(PBMCs).The expression of CX3CL1 and CCL26 in liver tissues were revealed by immuno-histochemistry.Using ELISA and flow cytometry,the changes of CX3CL1/CCL26-CX3CR1 pathway expression in human intrahepatic biliary epithelial cell(HIBEC)was studied upon stimulation of various cytokines and LPS.Results Forty patients with PBC and 18 HCs were included.The plasma levels of CX3CL1 were significantly higher in patients with PBC than HCs[(0.690±0.271)ng/mL vs.(0.540±0.101)ng/mL,P=0.044].An increased tendency was observed for plasma levels of CCL26 in patients with PBC compared with HCs[(8.94±4.09)pg/mL vs.(6.75±1.86)pg/mL,P=0.104].In comparison with HCs,the expression of CX3CR1 were significantly higher in NKT-like cells[(63.5±25.4)%vs.(78.6±18.0)%,P=0.026]and CD4^+T cells[(5.5±5.4)%vs.(13.7±9.0)%,P=0.003]in patients with PBC.Such differences existed in both CD28+and CD28-subsets of CD4^+T cells.The intrahepatic biliary epithelial cell in liver biopsy samples of PBC showed high expression of both CX3CL1 and CCL26.In contrast,HCs'only showed mild expression of CX3CL1 and no CCL26 expression.Upon stimulation of IFN-γ,HIBEC significantly increased its expression of CX3CL1 in intro,while IL-4 and IL-13 significantly increased the expression of CCL26.The expression of CX3CR1 was also up-regulated upon IFN-γstimulation.ConclusionsThe intrahepatic biliary epithelial cell in PBC may increase its expression of CX3CL1 upon stimulation of IFN-γ,and its expression of CCL26 upon IL-4 and IL-13.Meanwhile,their receptor CX3CR1 was up-regulated in peripheral NKT-like cells and CD4^+T cells.CX3C

关 键 词：原发性胆汁性胆管炎 CX3CL1-CX3CR1 CCL26 免疫机制 肝内胆管上皮细胞 

分 类 号：R57[医药卫生—消化系统]