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目的研究克罗恩病(CD)患者外周血白细胞介素(IL)-6水平与外周CD4~+CD25~+调节性T细胞(Treg细胞)频率及体外抑制功能的变化在CD发病中的作用。方法应用流式细胞术检测CD患者与正常对照者外周CD4~+CD25~+Treg细胞的频率及表型以及特征性标志叉状头/翅膀状螺旋转录因子(Foxp3)的表达,同时通过MACS缓冲液分选出外周血CD4~+CD25~+T细胞和CD4~+CD25~-T细胞,应用[~3H]-胸腺嘧啶渗入法研究CD4~+CD25~+T细胞对自体CD4~+CD25~-效应T细胞增殖的抑制能力。酶联免疫吸附法(ELISA)检测外周血IL-6水平。结果活动性CD患者外周血IL-6水平显著高于非活动性患者及正常对照者。活动性CD患者外周CD4~+T细胞中CD4~+CD25~+Foxp3~+T细胞的频率显著低于非活动性CD患者,差异有统计学意义。体外抑制功能试验同样提示活动性CD患者的CD4~+ CD25~+Treg细胞抑制功能减弱。结论CD4~+CD25~+Treg细胞抑制功能减弱与CD发病可能有关,可初步解释CD患者出现的免疫耐受缺失现象。
Objective To investigate the role of peripheral blood interleukin-6 (IL-6) and peripheral CD4 + CD25 + regulatory T cells (Treg cells) frequency and in vitro inhibition of Crohn’s disease (CD) in the pathogenesis of CD . Methods The frequency and phenotype of CD4 ~ + CD25 ~ + Treg cells and the expression of Foxp3, a characteristic marker of CD4 ~ + CD25 ~ + Treg cells in CD patients and normal controls, were detected by flow cytometry. CD4 ~ + CD25 ~ + T cells and CD4 ~ + CD25 ~ -T cells were isolated from the peripheral blood by using the method of [~ 3H] thymidine incorporation. Inhibition of effector T cell proliferation. Peripheral blood IL-6 levels were detected by enzyme-linked immunosorbent assay (ELISA). Results The levels of IL-6 in peripheral blood of patients with active CD were significantly higher than those of inactive patients and normal controls. The frequency of CD4 ~ + CD25 ~ + Foxp3 ~ + T cells in CD4 ~ + T cells in peripheral CD patients was significantly lower than that in inactive CD patients, the difference was statistically significant. In vitro inhibition of function tests also suggest that patients with active CD CD4 ~ + CD25 ~ Treg cell inhibition decreased. Conclusion The decreased CD4 ~ + CD25 ~ + Treg cells may play a role in the pathogenesis of CD, which may explain the lack of immune tolerance in CD patients.