Chen Hai-Ting, Wang Hong, Zhao Meng, Hou Bin
Beijing Tongren Eye Center,Beijing Tongren Hospital, Capital Medical University, Beijing 100730, China.
Zhonghua Yan Ke Za Zhi. 2012 Mar;48(3):234-40.
To assess the regulation of antigen specific Th17 cells differentiation in experimental autoimmune uveitis (EAU).
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-beta group, IL-6 group, IL-23 group, TGF-beta + IL-6 group, TGF-beta + 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.
Flow cytometric detection and ELISA experimental results showed:IRBP polypeptide alone mainly induced Th17 and Th1 response. Addition of TGF-beta and IL-6 induced the differentiation of antigen specific Th17 cells. Percentage of Th17 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 microg/L to 164.12 microg/L (t = -9.493, P = 0.009). Percentage of Th1 cells reduced from 6.33% to 3.43% (t = 6.059, P = 0.004). Percentage of Treg cells reduced from 4.96% to 1.52% (t = 5.683, P = 0.005); Furthermore, addition of IL-23 could enhance Th17 cells differentiation induced by TGF-beta and IL-6. Compared with IRBP polypeptide alone group, percentage of Th17 cells increased from 7.55% to 18.37% (t = -3.329, P = 0.029), and percentage of Th1 and Th2 cells reduced obviously (t = 7.410, P = 0.002; t = -3.863, P = 0.018). While IL-27 suppressed the differentiation of Th17 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, percentage of Th17 cells reduced from 7.55% to 4.06% (t = 2.163, P = 0.099).
Antigen specific Th17 differentiation is distinct from Th1 and Th2 cells. TGF-beta, IL-6 and IL-23 are the factors responsible for promoting the differentiation and development of Th17 subset, whereas IL-27 has inhibitory effects.
评估实验性自身免疫性葡萄膜炎（EAU）中抗原特异性Th17细胞分化的调控机制。
采用随机对照试验研究。通过在小鼠背部和腹部皮下注射光感受器间维生素A结合蛋白（IRBP）建立EAU模型。在第14天收集小鼠脾脏CD4 T细胞，在含有IRBP抗原的细胞培养板中于不同条件下进行体外培养72小时：对照组、转化生长因子-β（TGF-β）组、白细胞介素-6（IL-6）组、白细胞介素-23（IL-23）组、TGF-β + IL-6组、TGF-β + IL-6 + IL-23组、白细胞介素-27（IL-27）组、全反式维甲酸（ATRA）组。收集细胞和上清液。然后通过流式细胞术和酶联免疫吸附测定法评估Th17细胞、IL-17及其他细胞和细胞因子。所有数据采用学生t检验进行分析。
流式细胞术检测和酶联免疫吸附测定实验结果显示：单独的IRBP多肽主要诱导Th17和Th1反应。添加TGF-β和IL-6可诱导抗原特异性Th17细胞分化。Th17细胞百分比从7.55%增加至13.08%（t = -2.842，P = 0.048），细胞培养液中IL-17浓度从50.66μg/L增加至164.12μg/L（t = -9.493，P = 零009）。Th1细胞百分比从6.33%降至3.43%（t = 6.059，P = 0.004）。调节性T细胞（Treg）百分比从4.96%降至1.52%（t = 5.683，P = 0.005）；此外，添加IL-23可增强TGF-β和IL-6诱导的Th17细胞分化。与单独的IRBP多肽组相比，Th17细胞百分比从7.55%增加至18.37%（t = -3.329，P = 0.029），Th1和Th2细胞百分比明显降低（t = 7.410，P = 0.002；t = -3.863，P = 0.018）。而IL-27抑制Th17细胞分化。Th17细胞百分比从7.55%降至1.92%（t = 4.425，P = 0.041），而Treg细胞百分比从4.96%增加至9.98%（t = -5.073，P = 0.015）；在ATRA组中，Th17细胞百分比从7.55%降至4.06%（t = 2.163，P = 0.099）。
抗原特异性Th17分化不同于Th1和Th2细胞。TGF-β、IL-6和IL-23是促进Th17亚群分化和发育的因素，而IL-27具有抑制作用。
