树突状细胞负载Tα146-162治疗实验性自身免疫性重症肌无力小鼠的B细胞活化机制研究  

Effect of Dendritic Cells Pulsed with Tα146-162 on Autoimmune Myasthenia Graves and the Role of B Cells Activation

作　　者：王蓉[1] 杨欢[1] 肖波[1] 张丽芳[1] 

机构地区：[1]中南大学湘雅医院神经内科,长沙410008

出　　处：《四川大学学报（医学版）》2009年第5期793-797,共5页Journal of Sichuan University(Medical Sciences)

基　　金：国家自然科学基金(批准号30570639)资助

摘　　要：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 explore the therapeutic effect of Tαx146-162-iMDCs on C57BL/6 mice with EAMG and the role of changes in B cell activation regulated by Cbl. Methods Thirty four adult male C57BL/6 mice were randomly divided into EAMG group （A group）, EAMG group with interventions （B group） and control group （C group）. T-AChR antigens were injected to the mice in A and B groups to induce EAMG. Meanwhile, dendritic cells from immature bone marrows were cultured and pulsed with Tαx146-162, and injected to the mice in the intervention group. The mRNA expression of Cbl was detected by RT-PCR 90 days after the termination of experiment. The expression and phosphorylation of Syk and Lyn proteins were measured by Western blot. Results More mice （9/ 12, 75%） in A group developed EAMG than in B group （3/12, 25%, P〈0.05）. Mice in A group also suffered more serious illness in terms of clinical scores than mice in B group （1. 69±1.12 vs 0. 35±0.67, P〈0.01）. No EAMG occurred in mice in C group. Mice in B group had lower mRNA expressions of Cbl in the spleens and lymphonodes than mice in A groups, but both had higher expressions than mice in C group （P〈0.05）. Mice in B group had higher mRNA expression of Cbl than mice in A group （P〈0. 05）. Conclusion Tαx146-162-iMDCs prevent EAMG, possibly through negative regulation of Cbl on BCR signaling.

关 键 词：实验性自身免疫性重症肌无力 TΑ146-162 未成熟髓源性树突状细胞 CBL B细胞 

分 类 号：R746.1[医药卫生—神经病学与精神病学]