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

Expression and significance of OX40/OX40L in peripheral blood of patients with autoimmune hepatitis,primary biliary cholangitis,and their overlap syndrome

目的探讨自身免疫性肝炎(AIH)、原发性胆汁性胆管炎(PBC)及其重叠综合征患者经规范化治疗前后外周血中OX40/OX40L(CD134/CD134L)在CD4+T淋巴细胞、CD8+T淋巴细胞、单核细胞、B淋巴细胞的表达情况及其临床意义。方法选取2015年8月-2019年8月在江苏省苏北人民医院就诊的74例确诊AIH、PBC及其重叠综合征患者,按照相关诊断标准分为AIH组(29例)、PBC组(26例)、重叠综合征组(19例)(即A、P、C组),同时设置健康对照30例。在规范化治疗前后抽取患者外周血标本,采用免疫荧光标记流式细胞术检测OX40/OX40L在外周血细胞表面的表达情况,比较OX40/OX40L在治疗前后、健康对照组的表达差异。多组间比较采用单因素方差分析,进一步两两比较采用LSD-t检验,配对样本两组间比较采用配对t检验。结果3组在性别年龄构成上差异均无统计学意义(P值均>0.05)。治疗前:A、P、C 3组外周血中CD4+T淋巴细胞的OX40阳性率依次升高,且均高于对照组,依次为:(14.80±4.99)%、(17.11±2.71)%、(25.18±5.55)%、(6.67±2.26)%,差异有统计学意义(F=14.823,P<0.001);3组CD8+T淋巴细胞的OX40阳性率均高于对照组,依次为:(4.86±1.54)%、(6.40±1.88)%、(7.33±2.12)%、(4.09±2.69)%,差异有统计学意义(F=5.486,P<0.001);A、P、C 3组及对照组CD14+单核细胞的OX40L阳性率依次为(19.84±6.11)%、(21.17±4.35)%、(29.13±6.32)%、(4.86±2.34)%,差异有统计学意义(F=17.004,P<0.001);但在CD19+B淋巴细胞中,A、P、C 3组的OX40L阳性率却呈现逐渐降低趋势,但仍明显高于对照组,依次为(17.62±3.86)%、(14.75±4.32)%、(10.13±2.56)%、(4.50±1.38)%,差异有统计学意义(F=12.221,P<0.001)。治疗后:CD8+T淋巴细胞的OX40阳性率明显下降,与对照组水平接近,组间差异无统计学意义(F=0.731,P=0.538)。在其余3种细胞中,虽然OX40/OX40L的阳性率与治疗前相比较有不同程度的下降,但仍高于对照组:在CD4+T淋巴细胞中,A、P、C组OX40阳性率依次为(11.00±1.98)%、(13.72±1.03)%、(19.72±3.47)%,均高于对照组[(6.67±2.26)%](F=11.365,P<0.001);在CD14+单核细胞中,3种疾病的OX40L阳性率依次为(11.82±2.23)%、(15.19±4.42)%、(24.51±4.09)%,高于对照组[(4.86±2.34)%](F=13.748,P<0.001);在CD19+B淋巴细胞中,3种疾病的OX40L阳性率依次为(9.09±3.25)%、(6.81±2.20)%、(7.48±2.85)%,高于对照组[(4.50±1.38)%](F=8.052,P<0.001)。A、P、C 3组与治疗前OX40/OX40L在外周血CD4+T淋巴细胞、CD8+T淋巴细胞、CD14+单核细胞、CD19+B淋巴细胞的表达相比,治疗后均呈现明显下降,差异均有统计学意义(P值均<0.05)。结论OX40/OX40L在AIH、PBC及其重叠综合征患者外周血表达升高,经治疗后下降,表明OX40/OX40L通路参与了上述疾病发病机制,其中OX40在CD8+T淋巴细胞表面表达的作用或更能反映治疗效果。

Objective To 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.Methods A 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 difference t-test was used for further comparison between two groups;the paired t-test was used for comparison of paired samples between two groups.Results There 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(all P<0.05).Conclusion The 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.