结核分枝杆菌Rv1773c促进细菌侵入巨噬细胞的研究  

Mycobacterium tuberculosis Rv1773c promotes bacterial invasion of macrophages

作　　者：刘伟香 章晓联[1] LIU Wei xiang;ZHANG Xiao lian(Department of immunology, State Key Laboratory of Virology, Hubei Province Key Laboratory of Allergy and Immunology and Institute of Medical Research, Wuhan University School of Medicine , Wuhan 430071 , China)

机构地区：[1]武汉大学基础医学院免疫学系,病毒学国家重点实验室,湖北省过敏及免疫相关疾病重点实验室和医学研究院,湖北武汉430071

出　　处：《中国病原生物学杂志》2018年第4期330-335,共6页Journal of Pathogen Biology

基　　金：“十三五”国家重大传染病专项(No.2017ZX10201301-006); “十二五”国家重大传染病专项(No.2012ZX10003002-015)

摘　　要：目的筛选出能促进分枝杆菌对巨噬细胞侵入的功能性结核基因并进行验证。方法将H37Rv RD10-16区的18个结核分枝杆菌(MTB)基因构建到pMV261载体上,然后将重组pMV261质粒电转至耻垢分枝杆菌M.smegmatis(Ms)中,并将各重组菌株命名为rMs::Rvs。运用菌落计数法筛选功能性基因;采用流式细胞术Flow cytometry(FCM),动物实验验证该基因的功能。结果菌落计数显示rMs::Rv1773c组菌落数最多,与对照组相比差异有统计学意义(P<0.05);FCM验证Rv1773c促进Ms入侵巨噬细胞能力较强,rMs::Rv1773c组感染小鼠脾脏菌落数增多。结论筛选的RD14区功能性结核基因Rv1773c可能作为结核菌毒力因子,具备促进分枝杆菌入侵巨噬细胞的功能。Objective To identify the functional Mycobacterium tuberculosis genes that promote mycobacterial invasion of macrophages and to verify their identity using various experimental methods. Methods Eighteen M.tuberculosis H37 Rv ORFs in the RD10-16 region were cloned into the vector pMV261,and the recombinant plasmid pMV261 was used to transform M.smegmatis.Recombinant strains were designated rMs：：Rvs.The effects of these genes on mycobacterial invasion of macrophages were screened using colony counting and were further verified using flow cytometry（FCM）and a mouse model of M.smegmatis infection. Results The rMs：：Rv1773 chad the highest colony count among all groups in a colony count experiment,and the colony count differed significantly from that in the control group（P〈0.05）.FCM verified that Rv1773 cenhanced the ability of M.smegmatis to invade macrophages,and the number of bacterial colonies from the spleen of mice infected with rMs：：Rv1773 cincreased. Conclusion Rv1773 cfrom H37 Rv RD14 acted as a virulence factor,promoting the invasion of macrophages by mycobacteria.

关 键 词：结核分枝杆菌 入侵 巨噬细胞 RD区 菌落计数实验 Rv1773c 

分 类 号：R378.91[医药卫生—病原生物学]