摘要
目的:检测脾脏和淋巴结中B细胞活化相关蛋白表达及蛋白磷酸化表达水平的变化,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(EAMG的作用机制。方法:采用树突状细胞(DC)负载Tα146-162干预实验性自身免疫性重症肌无力小鼠(EAMG)的发病,34只6～8周龄健康雄性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DC,然后负载Tα146-162进行干预。从初次免疫起至第90天实验终止前,行EAMG严重性临床评估及发病率的计算。Western blot检测Syk、Lyn、Btk和PLC-γ2蛋白及蛋白磷酸化表达。结果:临床评估:A组发病率高于B组(75%vs25%,P<0.05)。实验终止时2组临床评分分别为1.69±1.12vs0.35±0.67(P<0.01)。C组小鼠无发病。A组小鼠的脾脏和淋巴结Syk和PLC-γ2蛋白表达和磷酸化水平较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);A组小鼠的脾脏和淋巴结Btk蛋白表达较C组小鼠升高(P<0.01),B组较A组降低(P<0.05),仍高于C组(P<0.05),但磷酸化水平三组无统计学意义(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,其机制可能与抑制B细胞活化有关。
AIM： To explore the therapic effect of Tα146-162-iMDCs in C57BL/6 mice with experimental autoimmune myasthenia graves（EAMG）, and illustrate whether this therapic effect is related with the change of B cells activation. METHODS. Adult male C57BL/6 mice were randomly divided into EAMG groups（A）, the prevention group （B） and control group（C） ; Immature bone marrow dendritic cells were cultured and pulsed with Tα146-162. Mice of A group and B group were evaluated for clinical score till the day they were put to death. The expression and phosphorylation of Syk, Lyn, Btk and PLC-γ2 protein were measured by Western blot. RESULTS： 75% mice of A group and 25% mice of B group were developed the accumulated incidence, the difference was significant （ P 〈 0.05 ）. The average clinical score of A group and B group were 1.69 ± 1. 12 vs 0.35±0.67（P〈0.01） at the termination of experiment. The expression and phosphorylation of Syk and PLC-γ2 protein in spleen and lymphonode of the mice of A group were higher than those of C group （P 〈 0.01） and B group （P 〈 0.05）. Compared with C group, those of B group were higher （ P 〈 0.05） ; The expression and phosphorylation of Lyn protein in spleen and lymphonode of the mice of A （ P 〈 0.01） and B （P 〈 0.05） groups were lower than those of C group. Compared with A group, those of B group were higher （ P 〈 0.05） ; The expression of Btk protein in spleen and lymphonode of the mice of A （ P 〈 0.01 ） and B （ P 〈 0.05） groups were higher than those of C group. And those of B groups were lower than those of A group （ P 〈 0.05）. But there were no remarkable differences among the phosphorylation of Btk protein of three groups. CONCLUSION： Tα46-162-iMDCs can prevent EAMG and probably ameliorate EAMG by the negative regulation on BCR signaling.
出处
《细胞与分子免疫学杂志》
CAS
CSCD
北大核心
2009年第1期42-45,共4页
Chinese Journal of Cellular and Molecular Immunology
基金
国家自然科学基金资助项目(30570639)
