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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。
Objective To investigate the expression and interaction of interleukin-10 (IL-10) and tumor necrosis factor-α (TNF-α) in BCG infected by human peripheral blood mononuclear cells (PBMCs) in vitro. Methods PBMCs were infected with BCG in vitro and the mRNA levels of IL-10 and TNF-α were detected by fluorescence quantitative PCR at 3, 6, 8 and 24 hours respectively. In addition, IL-10 antibody was incubated, the cell culture supernatants were collected and the contents of IL-10 and TNF-α in the supernatant were assayed by ELISA. Results Compared with the blank control group, the expression of IL-10 and TNF-α at the mRNA and protein levels increased after PBMCs were infected with BCG in vitro, the difference was statistically significant (P <0.05), the expression of both was negatively correlated When anti-IL-10 antibody was added, the level of TNF-α protein expression was higher than that of non-antibody group (P <0.05). Conclusion The expression of IL-10 and TNF-α in human PBMCs infected with BCG in vitro is increased, and the expression of TNF-α may be inhibited by IL-10.