诱导共刺激分子转基因小鼠类风湿性关节炎中滤泡辅助性T细胞的极化

目的 探讨可诱导共刺激分子(ICOS)转基因小鼠类风湿性关节炎(RA)模型中ICOS信号对滤泡辅助性T细胞(Tfh)极化的影响.方法 1.流式细胞仪分析测定ICOS转基因(ICOS-Trangenic,ICOS-Tg)小鼠及其野生型对照(WT)小鼠脾细胞CD4+T淋巴细胞共刺激分子ICOS在不同时期的表达趋势及特征;2.ELISA检测脾淋巴细胞培养悬液中干扰素(IFN)-γ、IL-21、IL-23、IL-17的水平;结果1.野生型RA小鼠脾CD4+T淋巴细胞表面的ICOS的表达水平从4w到12w为上升趋势(％)(4 W:6.5 ±1.0;7 W:13.2±1.3;12 W:23.5±:2.1);ICOS-Tg小鼠脾CD4+T淋巴细胞表面ICOS的表达三期同样呈上升趋势(％)(4W:8.2±0.9;7W:17.2±1.5;12W:31.6 ±3.0),但与野生型小鼠相比各期均表达上调(均P＜0.05);2.野生型小鼠IFN-γ从4W开始表达上升,7周达峰值后下降,ICOS-Tg小鼠同野生型小鼠相比,趋势相同但表达在各期呈下调趋势,差异有统计学意义(4 W～20 W均P＜0.05);野生型小鼠的IL-21、IL-23及IL-17的表达于4周上升,12周达峰值后下降,ICOS-Tg小鼠同野生型小鼠相比,三者的各期表达趋势相同但呈上调趋势(IL-21和IL-17有统计学意义,P＜0.05;IL-23无统计学意义,P＞0.05).结论 RA小鼠在其致病过程中共刺激信号ICOS呈上调表达;ICOS-Tg小鼠同野生型小鼠相比,其脾淋巴细胞培养上清中IFN-γ呈显著下调趋势;Tfh分化相关的细胞因子IL-21、IL-17则均显著上调表达.Tfh细胞及其作用因子很可能参与了RA的免疫应答,与RA的发生发展有关.

Tfh polarization of ICOS-Transgenic mice in Rheumatoid arthritis mediated by ICOS/ICOSL signaling pathway

Objective To explore the enhance of inducible costimulator(ICOS) signaling effection on the costimulatory molecules and cytokine expression levels associated with the polarization of Tfh in ICOS-Tg mice of RA.Methods 1.Determination of flow cytometric analysis of spleen cells CD4 + T cell costimulatory molecules ICOS;2 The detection of cytokines IFN-γ、IL-21 、IL-23 、IL-17 in cultured splenocytes suspension by ELISA;Results The expression of ICOS in splenic CD4 + T lymphocytes were gradually increased,peaking at 12 weeks,followed by a slow decline after 16 weeks remained at a high level of expression with the development of the course(4w:6.5 ± 1.0;7w:13.2 ± 1.3;12w:23.5 ± 2.1);compared to wild type mice,ICOS-Tg mice showed up-regulated expression of ICOS (4w:8.2 ± 0.9;7w:17.2 ± 1.5;12w:31.6 ± 3.0);the expression IL-21、IL-23、IL-17 of ICOS-Tg mice was significantly upregulated;the level of IFN-γ was downregulated compared with wild type.Conclusion Tfh cells may be involved in the immune response of RA.ICOS may be the key signaling on Tfh polarized immune responses.