【摘要】 目的研究外周血CD4+CD25+调节性T细胞(Treg细胞)表型和功能改变所致的细胞免疫抑制效应在复发性生殖器疱疹(RGH)发病机制中的作用。方法采用免疫荧光标染技术,经流式细胞仪检测34例RGH患者和33例正常人外周血CD4+CD25+T细胞转录因子Foxp3及细胞毒T细胞相关抗原4(CTLA-4)、糖皮质激素诱导的肿瘤坏死因子受体家族相关基因(GITR)、程序性死亡1(PD-1)等抑制性膜分子的表达水平。同时,用免疫磁珠细胞分选技术从外周血中分选出高纯度的CD4+CD25+T细胞,经体外刺激后分析胞内分泌白介素10(IL-10)和转化生长因子β(TGF-β)阳性细胞数。结果RGH患者外周血CD4+CD25+Foxp3+Treg细胞数量比正常人对照组显著增加(P<0.001),CD4+CD25+T细胞表面抑制性膜分子CTLA-4、GITR、PD-1表达明显增强(P分别<0.001、<0.001及<0.01),胞内IL-10和TGF-β阳性细胞增多(P值分别<0.01及<0.001)。结论单纯疱疹病毒感染可诱导CD4+CD25+Treg细胞活化增殖,高表达负性共刺激分子和分泌抑制性细胞因子,通过多种机制抑制机体的抗病毒免疫应答。更多还原

【Abstract】 Objective To investigate the phenotype and suppressive effects of peripheral blood CD4+CD25+ regulatory T cells (Tregs) on cell immunity in the pathogenesis of recurrent genital herpes (RGH). Methods The expression of transcription factor Foxp3 and inhibitory membrane molecules,e.g., cytotoxic T lymphocyte associated antigen-4(CTLA-4),glucocorticoid-induced TNF receptor family-related gene(GITR) and programmed death-1(PD-1),were determined by three-color flow cytometry in periph- eral CD4+CD25+T cells from 34 patients with RGH and 33 normal controls. Highly purified CD4+CD25+T cells were isolated ex vivo from peripheral blood by immunomagnetic beads, and the production of intracel- lular suppressor cytokines such as interleukin-10 (IL-10) and transforming growth factor-β(TGF-β) was tested by flow cytometry. Results The number of CD4+CD25+Foxp3+ Tregs increased significantly in pe- ripheral blood of patients with RGH (9.6%±2.4%) in comparison with that of normal controls(6.4%±2.1%) (P<0.001).The expression of CTLA-4, GITR and PD-1 was significantly stronger in patients (2.8%±1.3%,12.7%±2.5% and 1.9%±1.1%,respectively) than that in controls(1.3%±1.0%, 9.5%±2.4% and 1.3%±0.6%,respectively, P<0.001,P<0.001 and P<0.01,respectively).The numbers of IL-10- and TGF-β-positive CD4+CD25+T cells were significantly increased in peripheral blood from patients (2.24%±0.52% and 1.29%±0.42%, respectively) compared with those from controls (1.70%±0.53% and 0.73%±0.22%, respectively, P<0.01 and P<0.001,respectively).Conclu- sions Herpes simplex virus infection may result in the activation and proliferation of CD4+CD25+ Tregs, expression of high level negative costimulatory molecules on the cell surface, and secretion of suppressive cytokines, which may lead to the inhibition of antiviral immune responses via multiple mechanisms.更多还原