不同毒力钩端螺旋体引起THP-1细胞中细胞因子表达的研究  

Cytokine responses induced by Leptospira strains with different virulence in THP-1 cells

作　　者：张彦[1] 姜叙诚[2] 郭晓奎[1] 汤道强[3] 

机构地区：[1]上海交通大学医学院病原生物学教研室,上海200025 [2]上海交通大学医学院病理学教研室,上海200025 [3]上海交通大学医学院附属仁济医院,上海200127

出　　处：《微生物与感染》2011年第2期101-107,共7页Journal of Microbes and Infections

基　　金：上海市科学技术委员会项目(09XD1402700)

摘　　要：本文旨在通过观察不同毒力钩端螺旋体激活人单核巨噬细胞株THP-1中细胞因子信号途径的差异,探讨天然免疫在钩端螺旋体病发病机制中的作用。将3种不同毒力的钩端螺旋体——赖型有毒株Lai株、赖型减毒株IPAV株及非致病性Patoc型Patoc 1株与诱导后的细胞相互作用,在作用0、2、12、24、48 h后收集标本,应用实时聚合酶链反应(PCR)和酶联免疫吸附试验(ELISA)检测肿瘤坏死因子α(TNF-α)、γ干扰素(IFNγ-)、白细胞介素12(IL-12)和单核细胞趋化蛋白1(MCP-1)在基因和蛋白水平的表达,同时应用实时PCR检测细胞表面Toll样受体2(TLR2)、TLR4及诱导型一氧化氮合酶(iNOS)基因表达水平。结果显示,Lai株和IPAV株引起THP-1细胞中TLR4和iNOS基因表达升高,Patoc 1株引起THP-1细胞中TLR2、TLR4和iNOS基因表达升高。Lai株引起的THP-1细胞中TNF-α、MCP-1表达水平低于IPAV株和Patoc 1株(P<0.05)。Lai株引起较低水平细胞因子表达可能有利于其逃避宿主天然免疫反应,从而进一步在宿主体内播散、繁殖。The purpose of this study is to observe the cytokine responses induced by Leptospira strains with different virulence in THP-1 cells and to evaluate the role of innate immunity in the pathogenesis of leptospirosis.THP-1 cells induced by phorbol myristoyl acetate（PMA） were incubated with three different Leptospira strains,the virulent Leptospira interrogans serovar Lai type strain Lai,the avirulent Leptospira interrogans serovar Lai type strain IPAV,and the nonpathogenic Leptospira biflexa serovar Patoc type strain Patoc 1,respectively.Cells and supernatant infected by leptospires were collected at 0,2,12,24,and 48 h post-inoculation.The mRNA and protein levels of tumor necrosis factor α（TNF-α）,interferon γ（IFN-γ）,interleukin 12（IL-12）,and monocyte chemotactic protein 1（MCP-1） were quantified by real-time polymerase chain reaction（PCR） and enzyme-linked immunosorbent assay（ELISA）.Meanwhile,real-time PCR was used to detect mRNA levels of Toll-like receptor 2（TLR2）,TLR4,and inducible nitric oxide synthase（iNOS） in THP-1 cells infected by leptospires.It was found that Lai type strain Lai and Lai type strain IPAV could induce TLR4 and iNOS expression,but Patoc type strain Patoc 1 could induce TLR2,TLR4,and iNOS expression in THP-1 cells.TNF-α and MCP-1 levels in THP-1 cells inoculated with strain Lai were lower than those inoculated with strain IPAV and strain Patoc 1 for both mRNA and protein.Thus,the virulent strain Lai induces low 1evels of cytokines,which may facilitate escape of the host immune response and spreading.

关 键 词：钩端螺旋体 THP-1 TOLL样受体2 TOLL样受体4 细胞因子 

分 类 号：R377[医药卫生—病原生物学]