目的在细胞与分子水平检验重症肌无力(myasthenia gravis,MG)患者外周血中CD4^+CD25^+调节性T细胞(CD4^+CD25^+Tregs)的表达缺陷,探讨CD4^+CD25^+Tregs亚群异常与MG发病间的关系。方法流式细胞技术检测21例MG患者(11例经胸腺切除)与20名健康对照者(healthy controls,HCs)外周血CD4^+ CD25^+ Tregs及FoxP3^+CD4^+CD25^+Tregs含量,实时荧光定量聚合酶链反应(RT-FQ-PCR)分析MG患者与HCs外周血CD4^+CD25^+Tregs中FoxP3mRNA的表达。结果MG患者外周血CD4^+ CD25^+Tregs占CD4^+T细胞含量与HCs比较无统计学差异(P〉0.05)。MG患者外周血FoxP3+CD4^+CD25v+ Tregs含量及FoxP3mRNA表达量与HCs比较均显著性降低(P〈0.05);胸腺切除的MG患者与未经胸腺切除的MG患者外周血FoxP3^+CD4^+CD25^+Tregs含量及FoxP3 mRNA表达量无统计学差异(P〉0.05)。结论MG患者外周血CD4^+CD25^+Tregs数量正常,但其表面分子FoxP3的表达下调,这种CD4+CD25+Tregs亚群的异常发现有助于深入阐明MG的免疫发病机制。
Objective To identify the expression defects of CD4 ^+ CD25 ^+ regulatory T cells ( CD4 ^+ CD25 ^+ Tregs) in peripheral blood from myasthenia gravis (MG) patients at the cellular and molecular levels, with special focus on the effect of thymectomy on CD4 ^+ CD25 ^+ Tregs, and to investigate the relationship between the abnormalities of CD4 ^+ CD25 ^+ Tregs subset and the onset of MG. Methods Peripheral blood samples were obtained from 21 MG patients ( 11 patients have been thymectomized) and 20 healthy controls ( HCs), flow cytometry was used to analyze the number of CD4 ^+ CD25 ^+ Tregs and FoxP3 ^+ CD4 ^+ CD25 ^+ Tregs and real-time fluorescent quantitative ioolymerase chain reaction (RT-FQ-PCR) was used to determine the FoxP3 mRNA level of CD4 ^+ CD25 ^+ Tregs. Results There were no differences in the percentages of CD4 ^+ CD25 ^+ Tregs among CD4 ^+ T cells between MG patients and HCs ( P 〉 0. 05 ), the FoxP3-positive CD4 ^+ CD25 ^+ Tregs (FoxP3 ^+ CD4 ^+ CD25 ^+ Tregs) and the mRNA expression of FoxP3 were significantly different in CD4 ^+ CD25 ^+ Tregs of MG patients as compared to HCs ( P 〈 0. 05 ) ; the frequencies of CD4 ^+ CD25 ^+ Tregs and the expression of FoxP3 were similar in MG patients irrespective of treatment with thymectomy ( P 〉 0. 05 ). Conclusions The number of the CD4 ^+ CD25 ^+ Tregs is normal in MG patients, but the expression of their surface molecule Foxp3 is decreased. The abnormal finding on the CD4 ^+ CD25 ^+ Tregs subset could contribute to further elucidate the immunologic pathogenesis in MG.