Wang P, Song J, Fang X Y, Li X, Liu X, Jia Y, Li Z G, Hu F L
Department of Rheumatology and Immunology, Peking University People's Hospital & Beijing Key Laboratory for Rheumatism Mechanism and Immune Diagnosis, Beijing 100044, China.
State Key Laboratory of Natural and Biomimetic Drugs, Peking University School of Pharmaceutical Sciences, Beijing 100191, China.
Beijing Da Xue Xue Bao Yi Xue Ban. 2019 Jun 18;51(3):445-450. doi: 10.19723/j.issn.1671-167X.2019.03.011.
To explore the role of Ter cells in the development of the collagen-induced arthritis (CIA), we detected their quantity changes in the spleen of different stages of CIA mice and analyzed the correlation between Ter cells and the joint scores, and we also analyzed the correlation between Ter cells and the frequencies of T and B cell subsets, so as to further understand the pathogenesis of rheumatoid arthritis.
The six to eight weeks DBA/1 mice were used to prepare CIA model. After the second immunization, we began to evaluate the joint score. According to the time of CIA onset and the joint score, the CIA mice were divided into three stages: early, peak and late stages. According to the final joint score, the CIA mice at the peak stage were subdivided into the high score group (score>8) and the low score group (score≤8). The frequencies of Ter cells in the spleen of the naïve mice and the CIA mice at various stages and the frequencies of T and B cell subsets in the spleen of the CIA mice at the peak stage were detected by flow cytometry, then we carried on the correlation analysis.
The frequencies of Ter cells in the spleen of the CIA mice was significantly higher than those of the naïve mice (8.522%±2.645% vs. 1.937%±0.725%, P<0.01), the frequencies of Ter cells in the spleen of the high score group mice was significantly lower than those of the low score group (6.217%±0.841% vs. 10.827%±0.917%, P<0.01). The frequencies of Th1 cells in the spleen of the high score group mice was significantly higher than those of the low score group mice (1.337%±0.110% vs. 0.727%±0.223%, P<0.05). The frequencies of Th17 cells in the spleen of the high score group mice was higher than those of the low score group mice (0.750%±0.171% vs. 0.477%±0.051%, P=0.099). The frequencies of germinal center B cells in the spleen of the high score group mice was significantly higher than those of the low score group mice (1.243%±0.057% vs. 1.097%±0.015%, P<0.05). Correlation analysis results showed that the frequencies of Ter cells in the spleen of the CIA mice at the peak stage was strongly negatively correlated with the frequencies of CD4+ T, Th1, Th17, and germinal center B cells, and was strongly positively correlated with the frequencies of B10 cells, indicating that these cells might have a protective effect in CIA. Studies on dynamic changes showed that the frequencies of Ter cells in the spleen of the CIA mice at the late stage was significantly lower than those at the peak stage (0.917%±0.588% vs. 8.522%±2.645%, P<0.001), suggesting the protective effect of these cells in arthritis.
Ter cells were significantly increased in the spleen of the CIA mice at peak stage, and were negatively correlated with joint scores and pathogenic immune cells, and positively correlated with protective immune cells. Ter cells were significantly decreased in the spleen of the CIA mice at the late stage. What we mentioned above suggests that Ter cells might be involved in the progression of rheumatoid arthritis as an immunomodulatory cell,but further in vivo and in vitro experiments are needed to verify its specific effects and mechanism.
为探讨调节性T细胞(Ter细胞)在胶原诱导性关节炎(CIA)发病中的作用,我们检测了CIA小鼠不同阶段脾脏中Ter细胞数量的变化,并分析了Ter细胞与关节评分之间的相关性,同时还分析了Ter细胞与T、B细胞亚群频率之间的相关性,以进一步了解类风湿关节炎的发病机制。
选用6至8周龄的DBA/1小鼠制备CIA模型。第二次免疫后开始评估关节评分。根据CIA发病时间和关节评分,将CIA小鼠分为三个阶段:早期、高峰期和晚期。根据最终关节评分,将高峰期的CIA小鼠细分为高分组合(评分>8)和低分组合(评分≤8)。采用流式细胞术检测未免疫小鼠及各阶段CIA小鼠脾脏中Ter细胞的频率,以及高峰期CIA小鼠脾脏中T、B细胞亚群的频率,然后进行相关性分析。
CIA小鼠脾脏中Ter细胞的频率显著高于未免疫小鼠(8.522%±2.645% vs. 1.937%±0.725%,P<0.01),高分组合小鼠脾脏中Ter细胞的频率显著低于低分组合(6.217%±0.841% vs. 10.827%±0.917%,P<0.01)。高分组合小鼠脾脏中Th1细胞的频率显著高于低分组合小鼠(1.337%±0.110% vs. 0.727%±0.223%,P<0.05)。高分组合小鼠脾脏中Th17细胞的频率高于低分组合小鼠(0.750%±0.171% vs. 0.477%±0.051%,P=0.099)。高分组合小鼠脾脏中生发中心B细胞的频率显著高于低分组合小鼠(1.243%±0.057% vs. 1.097%±0.015%,P<0.05)。相关性分析结果显示,高峰期CIA小鼠脾脏中Ter细胞的频率与CD4+T、Th1、Th17和生发中心B细胞的频率呈强烈负相关,与B10细胞的频率呈强烈正相关,表明这些细胞可能在CIA中具有保护作用。动态变化研究表明,晚期CIA小鼠脾脏中Ter细胞的频率显著低于高峰期(0.917%±0.588% vs. 8.522%±2.645%,P<0.001),提示这些细胞在关节炎中具有保护作用。
高峰期CIA小鼠脾脏中Ter细胞显著增加,与关节评分和致病性免疫细胞呈负相关,与保护性免疫细胞呈正相关。晚期CIA小鼠脾脏中Ter细胞显著减少。上述结果提示,Ter细胞可能作为一种免疫调节细胞参与类风湿关节炎的进展,但需要进一步的体内和体外实验来验证其具体作用和机制。