白细胞介素-35在Vogt-小柳原田综合征患者中的表达及其对CD4^+CD25^+CD127dim/-调节性T细胞的调控作用  

The expression of IL-35 in patients with Vogt-Koyanagi-Harada syndrome and its modulatory effects to CD4^+CD25^+CD127dim/-regulatory T cells

作　　者：储昭节 王彤[1] 潘小燕 蔡敏 曲晓瑜 CHU Zhaojie;WANG Tong;PAN Xiaoyan;CAI Min;QU Xiaoyu(Department of Ophthalmology&Eye Institute of Shannxi Province,Xi’an First Hospital,Xi’an 710002,China)

机构地区：[1]西安市第一医院(西北大学附属第一医院)眼科西安市眼科医院陕西省眼科研究所,710002

出　　处：《免疫学杂志》2020年第6期533-538,共6页Immunological Journal

摘　　要：目的观察白细胞介素(IL)-35在Vogt-小柳原田(VKH)综合征患者外周血中的表达及其对CD4^+CD25^+CD127^dim/-调节性T细胞(Treg)功能的影响。方法收集37例VKH综合征患者(活动期16例,静止期21例)和11例健康对照者(healthy control,HC),分离血浆和外周血单个核细胞,酶联免疫吸附试验检测血浆IL-35水平,流式细胞术检测CD4^+CD25^+CD127^dim/-Treg比例。分选CD4^+CD25^+CD127^dim/-Treg,检测Treg分泌IL-35的水平和Treg中p35、EBI3 mRNA的相对表达量。使用重组人IL-35刺激分选的Treg后与自体CD4^+CD25-T细胞共培养,检测细胞增殖和分泌细胞因子的水平。结果活动期VKH综合征患者血浆IL-35水平低于静止期患者和HC(P<0.0001);活动期VKH综合征患者外周血CD4^+CD25^+CD127^dim/-Treg的比例(5.34±0.96%)也低于静止期患者和HC(P<0.01)。活动期VKH综合征患者分选的CD4^+CD25^+CD127^dim/-Treg分泌IL-35的水平(P=0.0036)以及Treg中p35和EBI3 mRNA的相对表达量(P<0.0001)均显著低于HC。重组人IL-35刺激不影响活动期VKH综合征患者CD4^+CD25^+CD127^dim/-Treg的增殖(P=0.323),但可增强Treg的免疫抑制功能,主要表现为共培养系统细胞增殖的下降(P=0.0005),抑制性细胞因子IL-10和IL-35的分泌增加(P<0.001),而促炎因子IFN-γ和TNF-α的水平降低(P<0.05)。活动期VKH综合征患者接受糖皮质激素治疗后,血浆IL-35水平和外周血CD4^+CD25^+CD127^dim/-Treg比例均显著升高(P<0.05)。结论IL-35和Treg在VKH综合征患者中水平降低,IL-35可调控Treg的免疫抑制功能,参与VKH综合征的发病。Vogt-Koyanagi-Harada(VKH)syndrome is an autoimmune disease affecting multiple systems.However,the pathogenesis of VKH syndrome is still not fully elucidated.Interleukin(IL)-35 has been revealed to play important regulatory roles in cancers,infections,and autoimmune disorders.Thus,the current study was designed to investigate the expression of IL-35 in patients with VKH syndrome,and to assess the modulatory activity of IL-35 to CD4^+CD25^+CD127^dim/-regulatory T cells(Treg).A total of 37 patients with VKH syndrome(including 16 of active and 21 of inactive)and 11 of healthy controls(HC)were enrolled in the study.Plasma and peripheral blood mononuclear cells(PBMC)were isolated;IL-35 expression in the plasma was measured by enzyme linked immunosorbent assay,while CD4^+CD25^+CD127^dim/-Treg percentage was investigated by flow cytometry.CD4^+CD25^+CD127^dim/-Treg was purified,and then IL-35 secretion by Treg and the mRNA relative levels of p35 and EBI3 were assessed.Purified Treg was stimulated with recombinant human IL-35,and then co-cultured with autologous CD4^+CD25-T cells.Subsequently,the cells and supernatants were harvested for cell proliferation assay and cytokine secretion analysis.Data showed that IL-35 expression in the plasma was significantly lower in active VKH syndrome patients when compared with inactive VKH syndrome patients and HC(P<0.0001),while peripheral CD4^+CD25^+CD127^dim/-Treg percentage was also notably down-regulated in active VKH syndrome patients(5.34±0.96%,P<0.01).The level of IL-35(P=0.0036)secreted from CD4^+CD25^+CD127^dim/-Treg as well as the relative mRNA levels of p35 and EBI3 in Treg(P<0.0001)were significantly decreased in active VKH syndrome patients when compared with HC.Recombinant human IL-35 stimulation did not affect the proliferation of CD4^+CD25^+CD127^dim/-Treg from patients with active VKH syndrome(P=0.323),however,significantly enhanced the immunosuppressive function of Treg,which presented as the reduction of cellular proliferation(P=0.0005),the elevation of inhibitory

关 键 词：细胞因子 CD4~^+T淋巴细胞 VOGT-小柳原田综合征 

分 类 号：R512.6[医药卫生—内科学]