胸腺基质淋巴细胞生成素诱导重症肌无力患者胸腺CD4^+CD25^-T细胞向CD4^+CD25^+Foxp3^+调节性T细胞分化  

Differentiation of Intrathymic CD4^+CD25^-T Cells to CD4^+CD25^+Foxp3^+ Regulatory T Cells Induced by Thymic Stromal Lymphopoietin in Myasthenia Gravis Patients

作　　者：卢祖能[1] 袁江[1] 张跃亮 王云甫[2] 

机构地区：[1]武汉大学人民医院神经内科,武汉430060 [2]湖北医药学院附属太和医院神经内科,湖北十堰442000

出　　处：《神经损伤与功能重建》2015年第2期134-137,共4页Neural Injury and Functional Reconstruction

基　　金：湖北省教育厅重点项目(No.D20112102);湖北省科技厅自然科学基金(No.2010CDB09103)

摘　　要：目的:探索胸腺基质淋巴细胞生成素(TSLP)对重症肌无力(MG)合并胸腺异常患者胸腺CD4+CD25-T细胞诱导生成CD4+CD25+Foxp3+调节性T(Treg)细胞的研究。方法:以磁珠分选的方法得到胸腺CD4+CD25-T细胞及树突状细胞(DC)细胞,通过不同浓度(0.1、1、10、20 ng/m L)的TSLP活化DC后诱导CD4+CD25-T细胞向CD4+CD25+Foxp3+Treg细胞分化,以流式细胞术检测CD4+CD25+Foxp3+Treg细胞/CD4+T细胞比率;RT-PCR检测Foxp3 m RNA表达;Western blot法检测上调的CD4+CD25+Foxp3+Treg细胞的Foxp3蛋白表达;ELSIA法检测培养上清IL-10的含量。结果:10 ng/m L TSLP组、20 ng/m L TSLP组经诱导后CD4+CD25+Foxp3+Treg细胞/CD4+T细胞比例均明显升高,Foxp3的表达水平也相应增加,IL-10水平升高,与PBS对照组相比差异有统计学意义(P<0.05)。结论:TSLP可以诱导MG合并胸腺异常患者胸腺CD4+CD25-T细胞向CD4+CD25+Foxp3+Treg细胞分化,并促进Foxp3的表达。Objective： To investigate the effect of thymic stromal lymphopoietin（TSLP） on the differentiation of intrathymic CD4^＋CD25^-T cells to CD4^＋CD25^＋Foxp3^＋regulatory T cells in myasthenia gravis（MG） patients with abnormal thymus gland. Methods： Intrathymic CD4^＋CD25^-T cells and dendritic cells isolated by immunomagnetic beads on Mini MACS device were co^-cultured with different concentrations of TSLP（0.1, 1, 10, 20 ng/mL） for 7days. The ratio of CD4^＋CD25^＋Foxp3^＋Treg/CD4^＋T cells was tested by flow cytometry. The expression of Foxp3 m RNA and protein was measured by RT-PCR and Western blot respectively. The concentration of serum IL-10 was detected by ELISA. Results： The ratio of CD4^＋CD25^＋Foxp3^＋Treg/CD4^＋T cell, the expression of Foxp3 m RNA and protein, and the level of IL-10 was increased significantly in groups with 10 ng/mL and 20 ng/mL TSLP compared with those in the PBS control group（P〈0.05）. Conclusion： TSLP may induce the differentiation of intrathymic CD4^＋CD25^-T cells to CD4^＋CD25^＋Foxp3^＋regulatory T cells in MG patients with abnormal thymus gland and up^-regulate the expression of Foxp3.

关 键 词：重症肌无力 胸腺基质淋巴细胞生成素 调节性T细胞 

分 类 号：R741[医药卫生—神经病学与精神病学] R746.1[医药卫生—临床医学]