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目的研究实验性自身免疫性重症肌无力(Experimental autoimmune myasthenia gravis,EAMG)大鼠发病过程中辅助性CD4+T细胞亚型的变化。方法将Lewis大鼠分为完全弗氏佐剂(CFA)早、晚期发病时对照相和EAMG早期(初次免疫后13d)、高峰期(初次免疫后50d)发病时相4组,各组分别给予相应的药物后,于发病早期和高峰期分离淋巴细胞,采用流式细胞术检测各组大鼠淋巴细胞中Th1、Th2、Th17和Treg细胞的比例,ELISA法检测淋巴细胞培养上清中各种细胞因子水平。结果在EAMG发病的早期时相,与CFA组比较,EAMG组Th1、Th2、Th17细胞的比例下降,而Treg细胞比例上升;淋巴细胞培养上清中IFNγ和IL-4水平显著下降(P<0.05),IL-6水平显著升高(P<0.05),IL-17和TGF-β水平略升高,但差异无统计学意义(P>0.05)。在EAMG发病的高峰期,与CFA组比较,EAMG组Th1和Th17细胞的比例显著上升(P<0.05),而Th2和Treg细胞的比例显著下降(P<0.05),淋巴细胞培养上清中IFNγ和IL-17水平显著上升(P<0.05),IL-4和TGF-β水平显著下降(P<0.01),IL-6水平略上升,但差异无统计学意义(P>0.05)。结论在EAMG发生过程中,4种辅助性CD4+T细胞亚型之间的平衡被打破,晚期时相中Th1和Th17细胞比例上升,而Th2和Treg细胞比例下降。
Objective To study the changes of helper CD4 + T cell subsets in the pathogenesis of experimental autoimmune myasthenia gravis (EAMG) in rats. Lewis rats were divided into complete Freund’s adjuvant (CFA) early and late onset of the control and EAMG early (13d after the initial immunization), the peak (50d after the initial immunization) phase 4 group, each group were given After corresponding drugs, lymphocytes were separated at the early and peak incidence stages. The proportion of Th1, Th2, Th17 and Treg cells in lymphocytes of each group was detected by flow cytometry. The levels of lymphocytes Cytokine levels. Results Compared with CFA group, the proportion of Th1, Th2 and Th17 cells in EAMG group decreased and the proportion of Treg cells increased in the early phase of EAMG. The levels of IFNγ and IL-4 in lymphocyte culture supernatant decreased significantly (P <0.05 IL-6 and IL-6 were significantly increased (P <0.05). The levels of IL-17 and TGF-β were slightly increased, but the difference was not statistically significant (P> 0.05). Compared with the CFA group, the proportion of Th1 and Th17 cells in EAMG group increased significantly (P <0.05), while the proportion of Th2 and Treg cells decreased significantly (P <0.05) in the peak period of EAMG onset. IFNγ in the supernatant of lymphocyte culture (P <0.05), IL-4 and TGF-βdecreased significantly (P <0.01), IL-6 level increased slightly, but the difference was not statistically significant (P> 0.05). Conclusion During EAMG, the balance between the four helper CD4 + T cell subtypes was disrupted. The proportion of Th1 and Th17 cells in the late phase increased, while the proportion of Th2 and Treg cells decreased.