论文部分内容阅读
目的从B细胞活化的角度探讨Tα146-162-iMDC干预实验性自身免疫性重症肌无力小鼠(EAMG)的作用机制。方法34只6～8周健康雄性C57BL/6J小鼠,随机分为模型组(A)、干预组(B)和对照组(C)。体外培养树突状细胞,然后负载Tα146-162对干预组小鼠进行干预。从初次免疫起至第90d实验终止前,行EAMG严重性临床评估及发病率的计算。RT-PCR法检测Cbl mRNA表达,Western blot法检测Syk和Lyn蛋白及蛋白磷酸化表达。结果A组发病率高于B组(75%vs25%,P<0.05)。实验终止时两组临床评分分别为1.69±1.12、0.35±0.67(P<0.01)。C组无发病小鼠。A组小鼠的脾脏和淋巴结Cbl mRNA表达水平均明显低于C组小鼠(P<0.01),B组小鼠的脾脏和淋巴结Cbl mRNA表达水平较A组明显升高(P<0.05),但仍低于C组(P<0.05)。A组小鼠的脾脏和淋巴结Syk蛋白和磷酸化表达水平较C组小鼠升高(P<0.01),B组较A组下降(P<0.05),但高于C组(P<0.05);A组小鼠的脾脏和淋巴结Lyn蛋白表达和磷酸化水平较C组小鼠降低(P<0.01),B组较A组升高(P<0.05),但仍低于C组(P<0.05)。结论Tα146-162-iMDC干预,能显著降低EAMG的发病率,改善临床症状,其机制可能与Cbl负性调控B细胞活化有关。
Objective To investigate the mechanism of Tα146-162-iMDC intervention in experimental autoimmune myasthenia gravis (EAMG) from the perspective of B cell activation. Methods Sixty-four healthy male C57BL / 6J mice aged 6-8 weeks were randomly divided into model group (A), intervention group (B) and control group (C). Dendritic cells were cultured in vitro, then Tα146-162 was loaded to intervene the intervention group. From the initial immunization to the 90th day before termination of the experiment, the clinical evaluation of EAMG severity and morbidity were performed. Cbl mRNA expression was detected by RT-PCR and Syk and Lyn protein and protein phosphorylation were detected by Western blot. Results The incidence of group A was higher than that of group B (75% vs 25%, P <0.05). At the end of the experiment, the clinical scores of the two groups were 1.69 ± 1.12,0.35 ± 0.67 (P <0.01). C group of mice without disease. The expression of Cbl mRNA in spleen and lymph node in group A was significantly lower than that in group C (P <0.01). The expression of Cbl mRNA in spleen and lymph node in group B was significantly higher than that in group A (P <0.05) But still lower than C group (P <0.05). Syk and phosphorylation of spleen and lymph node in group A were significantly higher than those in group C (P <0.01), but decreased in group B (P <0.05), but higher than those in group C (P <0.05) (P <0.01). The level of Lyn protein expression and phosphorylation in spleen and lymph node of group A were lower than that of group C (P <0.01), and the level of Lyn in group B was higher than that of group A (P < 0.05). Conclusion Tα146-162-iMDC intervention can significantly reduce the incidence of EAMG and improve clinical symptoms, the mechanism may be related to Cbl negative regulation of B cell activation.