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目的研究结核分枝杆菌主要分泌蛋白抗原85B(Ag85B)和卡介苗(BCG)对初治结核病患者树突状细胞(DC)免疫功能的调节作用。方法收集26例健康者及31例初治者,分离外周血单个核细胞。每个样本分成4组,诱导出DC并培养,在第6天对DC进行刺激,分别采用脂多糖(LPS)、BCG、Ag85B对3组细胞进行处理,不加刺激物处理的DC为对照组。处理24 h后收集细胞,流式细胞术检测DC的CD83、CD86、HLA-DR和CD11c水平。ELISA检测DC培养上清中的白细胞介素12(IL-12)、IL-10、γ干扰素(IFN-γ)水平,并用SPSS13.0软件对数据进行统计学分析。结果初治者未处理组中CD83及IL-10的表达量显著低于健康者未处理组,Ag85B组CD83、CD86和IFN-γ的含量显著高于未处理组,BCG组IFN-γ的含显著高于未处理组,LPS对照组CD83、CD86、HLA-DR和IL-10的含量显著高于未处理组。健康者LPS对照组CD83、CD86和IL-10的含量分别显著高于未处理组。结论 Ag85B对初治结核病患者DC的免疫增强功能优于BCG。
Objective To study the regulatory effect of Mycobacterium tuberculosis major secreted protein antigen 85B (Ag85B) and BCG on immune function of dendritic cells (DCs) in patients with newly diagnosed tuberculosis. Methods Peripheral blood mononuclear cells were collected from 26 healthy controls and 31 untreated patients. Each sample was divided into 4 groups, DCs were induced and cultured. DCs were stimulated on the 6th day. Three groups of cells were treated with lipopolysaccharide (LPS), BCG and Ag85B respectively. DCs without stimulus were control group . After 24 h of treatment, the cells were collected and the levels of CD83, CD86, HLA-DR and CD11c in DCs were detected by flow cytometry. The levels of IL-12, IL-10 and IFN-γ in DC culture supernatants were detected by ELISA. The data were analyzed by SPSS13.0 software. Results The expression levels of CD83 and IL-10 in untreated group were significantly lower than those in healthy group. The contents of CD83, CD86 and IFN-γ in Ag85B group were significantly higher than those in untreated group and IFN-γ in BCG group The levels of CD83, CD86, HLA-DR and IL-10 in the LPS control group were significantly higher than those in the untreated group. The levels of CD83, CD86 and IL-10 in healthy LPS control group were significantly higher than those in untreated group. Conclusions Ag85B is superior to BCG in immune enhancement of DC in patients with newly diagnosed tuberculosis.