布鲁菌基因缺失株△VceA的感染特征研究  

CHARACTERISTICS OF INFECTION OF BRUCELLA ΔVCEA DELETION STRAIN

作　　者：刘航[1] 何佳琪 王小凤 李明奇[1] 郭嘉 赵天艺 张樊 吴长新[1] 张辉[1] LIU Hang;HE Jia-qi;WANG Xiao-feng;LI Ming-qi;GUO Jia;ZHAO Tian-yi;ZHANG Fan;WU Chang-xin;ZHANG Hui(Key Laboratory of Xinjiang Endemic and Ethnic Disease,College of Animal Science&Technology,Shihezi University,Shihezi 832003,China)

机构地区：[1]石河子大学动物科技学院动物疾病防控兵团重点实验室,石河子832003

出　　处：《中国动物传染病学报》2020年第6期68-72,共5页Chinese Journal of Animal Infectious Diseases

基　　金：兵团人才项目(CZ027202,2017CB002);国际科技合作项目(GJHZ201709)

摘　　要：本文研究了布鲁菌IV型分泌系统效应蛋白VceA缺失对细胞侵染和小鼠感染特征。培养布鲁菌S2308基因缺失突变株ΔVceA,测定ΔVceA在生长过程中的OD值并制作其生长曲线;在用布鲁菌VceA基因缺失株和其亲本株S2308株分别侵染HPT-8细胞后,检测细胞炎性因子TNF-α和IL-1β的分泌。采用ΔVceA和亲本株S2308分别侵染HPT-8细胞和感染小鼠,对HPT-8进行胞内CFU计数,对小鼠脾脏内进行CFU计数,研究基因缺失突变株ΔVceA的感染特征。结果显示,基因缺失株ΔVceA和亲本株S2308的生长曲线在12 h达到对数生长期,30 h进入平台期;VceA基因缺失株在侵染细胞的初期胞内CFU计数低于亲本株,之后和亲本株无显著差异;基因缺失株ΔVceA在感染小鼠初期脾脏内CFU计数时低于亲本株,在感染后期和亲本株无明显差异;基因缺失株ΔVceA在侵染HPT-8细胞12 h时TNF-α和IL-1β的分泌量缺失株显著低于亲本株。结果表明,VceA基因的缺失不影响布鲁菌的生长繁殖,在布鲁菌感染细胞后影响细胞因子的分泌,胞内外的生存繁殖力明显下降,为深入研究布鲁菌IV型分泌系统的作用和功能奠定基础。To study the function of effector protein VceA of Brucella T4SS,Brucella S2308 gene deletion mutant ΔVceA was cultured and its growth was measured in OD values.The mutant VceA strain and its parent strain were used to infect HPT-8 cells and mice for examination of the release of cytokines TNF-α and IL-1β in the cell and CFU counts in HPT-8 cells and mouse spleens.The results showed that the growth curves of the mutant VceA strain and its parent reached to the logarithmic growth stage at 12 hours and to the plateau stage at 30 hours post-infection.The CFU counts of the mutant VceA strain was lower than that of its parent at the initial infection stage for HPT-8 cells and mice.In addition,the secretion of TNF-α and IL-1β was not significantly different in HPT-8 cells at 12 hours post-infection with the mutant VceA strain and its parent.However,the loss rate of the mutant VceA strain was significantly lower than that of parents (P<0.05).The results showed that the deletion of VceA gene did not affect the growth and reproduction of Brucella but affected the secretion of cytokines after infection.The significant decrease of intracellular and extracellular viabilities was worthwhile for further study of the Brucella T4SS.

关 键 词：布鲁菌 VceA 生长曲线 HPT-8细胞 

分 类 号：S852.614[农业科学—基础兽医学]