结核病患者外周血和胸腔积液中多功能Th1细胞检测

Detection of multifunctional Th1 cells in peripheral blood and pleural effusion in TB patients

目的探讨结核病患者外周血和胸腔积液中IL-2+IFN-γ+TNF-α+多功能Th1细胞的水平及其临床意义。方法选取结核患者49例,其中结核性胸膜炎患者14例,结核潜伏感染者27例,健康对照66例,采用佛波酯(PMA)和离子霉素刺激全血和胸腔积液中单个核细胞,胞内细胞因子染色,流式细胞术检测CD4+T细胞细胞因子分泌的情况。结果依据CD4+T细胞产生细胞因子的不同,将其分为7个不同亚群,分别为IL-2+IFN-γ+TNF-α+、IL-2+IFN-γ+、IL-2+TNF-α+、IFN-γ+TNF-α+、IL-2+、TNF-α+、IFN-γ+细胞亚群,外周血中结核病患者的IL-2+IFN-γ+TNF-α+多功能Th1细胞的比例明显低于健康对照和潜伏感染者(P<0.01),IL-2+IFN-γ+细胞、IFN-γ+TNF-α+细胞表达水平明显低于潜伏感染者和健康对照(P<0.05);TNF-α+细胞则高于健康对照和潜伏感染者(P<0.05);其他各亚群无明显变化。胸腔积液单个核细胞(PEMC)的IL-2+IFN-γ+TNF-α+多功能Th1应答水平高于外周血单个核细胞(P<0.05);外周血单个核细胞中IL-2+细胞低于PEMC(P<0.01),其他各亚群无明显变化。结论非特异性Th1细胞应答与结核菌感染的临床转归有关,IL-2+IFN-γ+TNF-α+多功能Th1细胞在结核的保护性免疫反应中起一定作用。

Objective To investigate the level of IL-2+ IFN-γ+ TNF-α+ multifunctional Th1 cell in peripheral blood and hydro-thorax of the TB patients and its clinical significance .Methods 49 patients with tuberculosis(TB) including 14 cases of tuberculous pleurisy and 27 individuals with latent TB infection were selected and 66 healthy individuals were selected as the controls .PMA and ionomycin were adopted to stimulate mononuclear cells in whole blood and pleural effusion .The secretion status of CD4+ T cells cy-tokines was detected by using the intracellular cytokine staining and the flow cytometric analysis .Results According to the differ-ent cytokines generated by CD4+ T cells ,which were divided into 7 cell subgroups :IL-2+ IFN-γ+ TNF-α+ ,IL-2+ IFN-γ+ ,IL-2+TNF-α+ ,IFN-γ+ TNF-α+ ,IL-2+ ,TNF-α+ and IFN-γ+ cell subgroups .The proportion of peripheral blood IL-2+ IFN-γ+ TNF-α+multifunctional Th1 cells in the TB patients was significantly lower than that in the healthy controls and the individuals with latent TB infection(P<0 .01) ,the expression levels of IL-2+ IFN-γ+ cells and IFN-γ+ TNF-α+ cells were significantly lower than those in the individuals with latent TB infection and the healthy controls (P<0 .05);TNF-α+ cells was higher than that in the healthy con-trols and the individuals with latent TB infection (P<0 .05) .The other subgroups had no obvious change .The response level of IL-2+ IFN-γ+ TNF-α+ multifunctional Th1 cells in the pleural effusion mononuclear cells (PEMC) was higher than that in the peripher-al blood mononuclear cells(P<0 .05);IL-2+ cells in peripheral blood mononuclear cells (PBMC) was lower than that in PEMC (P<0 .01) .Conclusion The response of non-specific Th1 cells is related with the clinical outcome of TB infection ,IL-2+ IFN-γ+TNF-α+ multifunctional Th1 cells plays a certain role in the protective immunoreaction of TB .