实验性自身免疫性葡萄膜炎中抗原特异性CD4＋辅助性T17细胞的分化调控  

Regulation of antigen specific Th17 cells differentiation in experimental autoimmune uveitis

作　　者：陈海婷[1] 王红[1] 赵萌[1] 侯彬[1] 

机构地区：[1]首都医科大学附属北京同仁医院北京同仁眼科中心北京市眼科学与视觉科学重点实验室,100730

出　　处：《中华眼科杂志》2012年第3期234-240,共7页Chinese Journal of Ophthalmology

基　　金：国家自然科学基金（30772013）

摘　　要：目的探讨实验性自身免疫性葡萄膜炎（EAU）中影响抗原特异性CD4^＋辅助性T（Th）17细胞分化的调控因素。方法随机对照实验研究。向小鼠背部及胁腹部多点皮下注射光受体视黄醇类结合蛋白（IRBP）制作EAU模型，提取小鼠致敏14d的脾脏CD4^＋T细胞，置于含有IRBP抗原的细胞培养板中，加入不同组合的调控细胞因子，分为8组：对照组、转化生长因子（TGF）-β组、白细胞介素（IL）-6组、IL-23组、TGF—B＋IL-6组、TGF-p＋IL-6＋IL-23组、IL-27组和全反式维A酸（ATRA）组。培养72h，收集细胞与上清液，通过流式细胞仪与酶联免疫吸附试验检测Th17细胞的分化及细胞因子的水平。利用独立样本单因素t检验分析数据。结果流式细胞仪和酶联免疫吸附实验检测结果显示，TGF—β和IL-6共同存在的条件下，Th17细胞的百分比由7．55％升至13．08％（t=-2．842，P=0．048），细胞培养上清液中IL-17的浓度由50．66μg/L升至164．12μg/L（t=-9．493，P=0．009），Th-细胞的百分比由6．33％降至3．43％（t=6．059，P=0．004），调节性T细胞的百分比由4．96％降至1．52％（t=5．683，P=0．005）；IL-23进-步促进了TGF-β和IL-6诱导的Th17细胞分化，与单纯极化条件培养组相比，Th17细胞的百分比由7．55％升至18．37％（t=～3．329，P=0．029），Th1细胞（t=7．410，P=0．002）、Th2细胞（t=-3．863，P=0．018）百分比均明显降低；而IL-27组Th17细胞的百分比由7．55％降至1．92％（t=4．425，P=0．041），Th1细胞百分比由6．33％降至0．63％（t=13．847，P=0．001），而调节性T细胞百分比由4．96％升至9．98％（t=-5．073，P=0．015），ATRA组Th17细胞的百分比由7．55％降至4．06％（t=2．163，P=0．099）。结论抗原特异性Th17细胞的分化是不同于Th1和Th2细胞的独立的分化过程，TGF—β、IL-6和IL-23可诱导或促进Th17细朐的分化．而IL-27抑制�Objective To assess the regulation of antigen specific Thl7 cells differentiation in experimental autoimmune uveitis （EAU）. Methods A randomized controlled trials research. EAU model was made through subcutaneous injection of interphotoreceptor retinoid-binding protein（IRBP） at backs and bellies. Mouse splenic CD4^＋ T cells were collected on the fourteenth day and cultured in vitro in the cell culture plates containing IRBP antigen for 72 hours under different conditions：control group,TGF-β group, IL-6 group, IL-23 group ,TGF-β ＋ IL-6 group ,TGF-β ＋ IL-6 ＋ IL-23 group, IL-27 group, all transretinoic acid （ATRA） group. Cells and the clear liquid were collected. Then Th17 cells, IL-17 and other cells, cytokines were assessed by flow cytometry and ELISA. All data were analyzed by Student＇s t test. Results Flow cytometric detection and ELISA experimental results showed： IRBP polypeptide alone mainly induced Thl7 and Thl response. Addition of TGF-[5 and IL-6 induced the differentiation of antigen specific Thl7 cells. Percentage of Tb 17 cells increased from 7.55 % to 13.08 % （ t = - 2. 842, P = 0.048 ） , and the concentration of IL-17 in cell culture fluid increased from 50. 66 μg/L to 164. 12 μg/L （ t = - 9. 493, P = 0. 009 ）. Percentage of Thl cells reduced from 6.33% to 3.43% （ t = 6. 059, P = 0. 004）. Percentage of Treg cells reduced from 4.96% to l. 52% （t = 5. 683 ,P = 0. 005） ;Furthermore, addition of IL-23 could enhance Th17 ceils differentiation induced by TGF-13 and IL-6. Compared with IRBP polypeptide alone group, percentage of Th17 ceils increased from 7.55% to 18.37% （t = - 3. 329,P =0. 029） ,and percentage of Thl and Th2 cells reduced obviously（ t = 7. 410, P = 0. 002; t = - 3. 863, P = 0. 018 ）. While IL-27 suppressed the differentiation of Thl7 cells. Percentage of Th17 cells reduced from 7.55% to 1.92% （t = 4. 425, P = 0. 041 ） ,while percentage of Treg cells increased from 4.96% to 9.98% （t = -5. 073, P = 0. 015 ） ;In ATRA group, percenta

关 键 词：T淋巴细胞 辅助诱导 自身免疫疾病 葡萄膜炎 细胞因子类 

分 类 号：R773[医药卫生—眼科]