IL-10和TNF-α在人外周血单个核细胞体外感染卡介苗中的表达及相关性研究

目的探讨白细胞介素-10(IL-10)、肿瘤坏死因子(TNF-α)在人外周血单个核细胞(PBMCs)体外感染卡介苗(BCG)中的表达及相互作用。方法 PBMCs体外感染BCG后,分别于3、6、8、24h收集细胞应用荧光定量PCR法检测细胞内IL-10和TNF-α的mRNA含量;此外,分别于8、20、32、48h加入anti-IL-10的抗体孵育,收集细胞培养上清,用ELISA法检测上清中IL-10和TNF-α的蛋白含量。结果 PBMCs体外感染BCG后,相对于空白对照组,IL-10和TNF-α在mRNA和蛋白水平的表达量均增高,差异具有统计学意义(P<0.05),两者表达量呈现负相关趋势;当加入anti-IL-10的抗体后,TNF-α的蛋白表达量要高于未加抗体组,差异具有统计学意义(P<0.05)。结论 IL-10和TNF-α在人PBMCs体外感染卡介苗中的表达量增加,IL-10对TNF-α的表达可能具有抑制作用。

Expression of cytokine interleukin-10 and tumor necrosis factor alpha in human peripheral blood monouclear cells infected with bacille calmette-guerin

Obiective To investigate the expression of IL-10 and TNF-α in peripheral blood monouclear cells(PBMCs) infected with bacille calmette-guerin(BCG).Methods The mRNA levels of IL-10 and TNF-α in the PBMCs infected with BCG for 3 h,6 h,8 h and 24 h were determined using Real-time Quantitative PCR(RT-PCR).Culture supernatant samples were collected at 8 h,20 h,32 h and 48 h of the PBMCs treated with anti-IL-10 antibody.The levels of IL-10 and TNF-α in the supernatant were determined by enzyme-linked immunosorbent assay(ELISA).Results IL-10 and TNF-α levels released by PBMCs significantly increased after stimulation by BCG in mRNA and protein expression levels(P<0.05).A significantly negative regression was found between them.TNF-α levels were significantly higher than the control group after adding exogenous anti-human-IL-10(P<0.05).Conclusion BCG could induce the expression of IL-10 and TNF-α in PBMCs,and IL-10 could inhibit the productions of TNF-α.The result provided a basis for study of new type anti-tuberculosis vaccines and tuberculosis immunological pathogenesis.