自身免疫性肝炎、原发性胆汁性胆管炎及其重叠综合征患者外周血OX40/OX40L的表达及意义

王维钊， 朱沁玲， 向晓星， 马 莉， 顾 达， 童 聪， 王婷婷，s何家俊， 刘 军， 王 璐, 陈超伍

1 江苏省苏北人民医院， 扬州大学临床医学院 a.内镜诊治中心； b.消化内科； c.风湿免疫检测中心，江苏 扬州 225001； 2 大连医科大学 研究生院， 辽宁 大连 116044

ExpressionandsignificanceofOX40/OX40Linperipheralbloodofpatientswithautoimmunehepatitis,primarybiliarycholangitis,andtheiroverlapsyndrome

WANGWeizhao1a,ZHUQinling1b,2,XIANGXiaoxing1b,MALi1b,GUDa1b,TONGCong1b,WANGTingting1b,2,HEJiajun1b,2,LIUJun1a,WANGLu1a，CHENChaowu1a.

(1.a.CenterofEndoscopicDiagnosisandTreatment,b.DepartmentofGastroenterology,c.CenterofRheumatologyandImmunology,SubeiPeople’sHospitalofJiangsu&ClinicalMedicalCollegeofYangzhouUniversity,Yangzhou,Jiangsu225001,China;2.GraduateSchool,DalianMedicalUniversity,Dalian,Liaoning116044,China)

Abstract:ObjectiveTo investigate the expression and clinical significance of OX40/OX40L (CD134/CD134L) in CD4+T cells, CD8+T cells, monocytes, and B lymphocytes in peripheral blood of patients with autoimmune hepatitis (AIH), primary biliary cholangitis (PBC), and their overlap syndrome before and after standardized treatment.MethodsA total of 74 patients with AIH, PBC, and their overlap syndrome who were diagnosed in Subei People’s Hospital of Jiangsu from August 2015 to August 2019 were enrolled, and according to related diagnostic criteria, they were divided into AIH group (group A) with 29 patients, PBC group (group P) with 26 patients, and overlap syndrome group (group C) with 19 patients. A healthy control group with 30 individuals was also established. Peripheral blood samples were collected before and after standardized treatment to measure the expression of OX40/OX40L on the surface of peripheral blood cells by immunofluorescence flow cytometry, and the expression of OX40/OX40L was compared before and after treatment and between the three groups and the healthy control group to investigate its clinical significance. A one-way analysis of variance was used for comparison between multiple groups, and the least significant differencet-test was used for further comparison between two groups; the pairedt-test was used for comparison of paired samples between two groups.ResultsThere were no significant differences in sex composition and age composition between the three groups (P>0.05). Before treatment, the positive rate of OX40 in peripheral blood CD4+T cells gradually increased in groups A, P, and C, and groups A, P, and C had a significantly higher positive rate of OX40 than the control group (14.80%±4.99%/17.11%±2.71%/25.18%±5.55% vs 6.67%±2.26%,F=14.823,P<0.001); groups A, P, and C had a significantly higher positive rate of OX40 in CD8+T cells than the control group (4.86%±1.54%/6.40%±1.88%/7.33%±2.12% vs 4.09%±2.69%,F=5.486,P<0.001); the positive rate of OX40L in CD14+monocytes was 19.84%±6.11% in group A, 21.17%±4.35% in group P, 29.13%±6.32% in group C, and 4.86%±2.34% in the control group, and there was a significant difference between groups (F=17.004,P<0.001); the positive rate of OX40L in CD19+B cells was 17.62%±3.86% in group A, 14.75%±4.32% in group P, 10.13%±2.56% in group C, and 4.50%±1.38% in the control group, showing a trend of gradual reduction, and groups A, P, and C had a significantly higher positive rate than the control group (F=12.221,P<0.001). After treatment, the positive rate of OX40 in CD8+T cells decreased significantly to a similar level as the control group, and there was no significant difference between groups (F=0.731,P=0.538). For the other three types of cells, although there were varying degrees of reduction in the positive rate of OX40/OX40L after treatment, groups A, P, and C still had a significantly higher positive rate than the control group; in CD4+T cells, the positive rate of OX40 was 11.00%±1.98% in group A, 13.72%±1.03% in group P, 19.72%±3.47% in group C, and 6.67%±2.26% in the control group, and groups A, P, and C had a significantly higher positive rate than the control group (F=11.365,P<0.001); in CD14+monocytes, the positive rate of OX40L was 11.82%±2.23% in group A, 15.19%±4.42% in group P, 24.51%±4.09% in group C, and 4.86%±2.34% in the control group, and groups A, P, and C had a significantly higher positive rate than the control group (F=13.748,P<0.001); in CD19+B cells, the positive rate of OX40L was 9.09%±3.25% in group A, 6.81%±2.20% in group P, 7.48%±2.85% in group C, and 4.50%±1.38% in the control group, and groups A, P, and C had a significantly higher positive rate than the control group (F=8.052,P<0.001). Groups A, P, and C had significant reductions in the expression of OX40/OX40L in peripheral blood CD4+T cells, CD8+T cells, CD14+monocytes, and CD19+B lymphocytes after treatment (allP<0.05).ConclusionThe expression of OX40/OX40L in peripheral blood increases in patients with AIH, PBC, and their overlap syndrome and decreases after treatment, indicating that the OX40/OX40L pathway is involved in the pathogenesis of the above diseases, and the role of OX40 on the surface of CD8+T cells may better reflect the treatment outcome.

Keywords：hepatitis, autoimmune; primary biliary cholangitis; overlap syndrome; receptors, OX40; OX40 ligand

自身免疫性肝病是一组自身免疫功能紊乱导致的以肝脏为靶器官损害的自身免疫病，主要包括自身免疫性肝炎(AIH)、原发性胆汁性胆管炎(PBC)、原发性硬化性胆管炎(PSC)，以及其重叠综合征，多见于女性。其中AIH、PBC、AIH重叠PBC较为常见，目前此三种疾病发病机制尚不完全明确，当前的研究认为其发病与免疫紊乱、遗传、环境因素相关。T、B淋巴细胞的异常激活，及单核巨噬系统的活化，导致T淋巴细胞功能亢进，B淋巴细胞异常分泌自身抗体，单核巨噬细胞系统分泌炎性细胞因子，参与了其发病。而T淋巴细胞的活化需要第一与第二协同刺激信号及专职抗原递呈细胞的参与[1-3]。OX40 (CD134)及其配体OX40L(CD252、CD134L)是肿瘤坏死因子受体超家族的成员，它们的结合能为上述细胞活化提供协同刺激信号，其中，OX40表达于活化的CD4+、CD8+T淋巴细胞表面，OX40L主要表达于活化的B淋巴细胞、单核细胞、巨噬细胞。OX40/OX40L信号通路结合活化后，能促进CD4+T淋巴细胞迁移、增殖、分化和记忆性CD4+T淋巴细胞的产生，增强CD8+T淋巴细胞功能，而迁移的CD4+T淋巴细胞可辅助B淋巴细胞的成熟及自身抗体的产生，OX40/OX40L与多种自身免疫病的发生发展密切相关[4-7]，但目前尚缺乏OX40/OX40L在AIH、PBC及其重叠综合征作用的相关研究，本研究通过检测OX40/OX40L在上述3种疾病患者外周血淋巴单核细胞的表达，探讨其临床意义。