TLR2及其相关细胞因子在军团菌感染中作用  

Role of TLR2 and its related cytokines in Legionella infection

作　　者：张帆[1] 王素萍[1] 高向东[1] 邬惟为 董洁敏[1] 

机构地区：[1]山西医科大学公共卫生学院流行病学教研室

出　　处：《中国公共卫生》2015年第4期441-443,共3页Chinese Journal of Public Health

基　　金：国家自然科学基金(30872154);高等学校博士学科点专项科研基金(20040114004)

摘　　要：目的通过体外实验探讨Toll样受体2(TLR2)及肿瘤坏死因子-α(TNF-α)、白介素1β(IL-1β)、白介素6(IL-6)在军团菌感染中的作用。方法分离30名健康人外周血单核细胞(PBMC),将每名健康人PBMC分为对照组和军团菌刺激组,军团菌刺激组加入500μL 2×107/m L军团菌悬液,对照组加入500μL磷酸盐缓冲液(PBS),分别在18、24、72 h时获取细胞及上清液,采用real-time PCR法测定细胞中TLR2、My D88 mRNA的表达水平,ELISA法检测细胞培养上清液IL-6、IL-1β及TNF-α含量。结果与对照组比较,18h军团菌刺激组My D88 mRNA△Ct值(1.414 0±0.044 9)低于对照组(1.684 4±0.042 6),24h军团菌刺激组TLR2 mRNA△Ct值(1.347 9±0.056 2)低于对照组(1.425 9±0.038 7),72h军团菌刺激组TLR2 mRNA和My D88 mRNA△Ct值[(1.504 9±0.018 9)和(1.540 0±0.031 9)]均高于对照组[(1.410 5±0.025 5)和(1.339 2±0.032 2)],差异均有统计学意义(均P<0.05);18、24、72h军团菌刺激组细胞上清液中TNF-α、IL-1β、IL-6浓度均高于对照组,差异均有统计学意义(均P<0.05)。结论 TLR2及TNF-α、IL-1β、IL-6等相关炎性因子可有效抵制军团菌感染,TLR2介导的天然免疫可能是军团菌感染的保护因素。Objective To explore the role of Toll like receptor 2（TLR2） and its related cytokines in Legionella infection by in vitro experiment. Methods Peripheral blood mononuclear cells（PBM Cs） were obtained from healthy subjects（n = 30） and were stimulated with live Legionella pneumophila（Lp） for 18,24,and 72 hours. The mRNA expression levels of TLR2 and myeloid differentiation factor 88（M y D88） in PBM Cs were determined using real-time reverse transeription PCR（RT-PCR）. The expression levels of tumor necrosis factor-α（TNF-α）,interleukin-1β（IL-1β）,and interleukin-6（IL-6） were measured with enzyme-linked immunosorbent assay（ELISA）. Results TLR2 mRNA expression level was not significantly different betw een stimulation group and control group in 18 hours（P〉0. 05）. TLR2 mRNA expression level of stimulation group was significantly higher than that of control group（1. 3479 ± 0. 0562 vs. 1. 4259 ±0. 0387,P〈0. 05） in 24 hours. The My D88 mRNA expression level of stimulation group was significantly higher than that of control group（1. 4140 ± 0. 0449 vs. 1. 6844 ± 0. 0426,P〈0. 05） in 18 hours. The My D88 mRNA expression level was not statistically significantly different betw een stimulation group and control group in 24 hours（P〉0. 05）. TLR2 mRNA and My D88 mRNA expression levels of stimulation group were significantly low er than those of control group（P〈0. 05） in 72 hours. The concentration of TNF-α,IL-1β and IL-6 for Legionella stimulation group were significantly higher than those of the control group（P〈0. 05） in 18,24 and 72 hours. Conclusion Our study show ed that TLR2 and its related inflammatory cytokines TNF-α,IL-1β and IL-6 can effectively resist Legionella infection and TLR2 mediated innate immunity may be a protective factor for Legionella infection.

关 键 词：Toll样受体2(TLR2) 肿瘤坏死因子-α(TNF-α) 白介素1β(IL-1β) 白介素6(IL-6) 军团菌 感染 

分 类 号：R563[医药卫生—呼吸系统]