黑龙江畜牧兽医Issue(1)：21-29,9.DOI:10.13881/j.cnki.hljxmsy.2024.11.0104
Rv1140蛋白酶在结核分枝杆菌感染中的作用研究
Study on the role of Rv1140 protease in Mycobacterium tuberculosis infection
摘要
Abstract
To further elucidate the function of Rv1140 protease in Mycobacterium tuberculosis(MTB)infection,the study employed homologous recombination to construct MTB H37Ra rv1140 gene knockout strains(ΔRv1140 strains)and MTB H37Ra rv1140 gene complementation strains(ΔRv1140+Rv1140 strains),which were identified via PCR and Southern-blot analysis.Growth curves were plotted for the H37Ra,ΔRv1140,and ΔRv1140+Rv1140 strains.Following infection of macrophages with the H37Ra,post infection rv1140 transcription levels were detected using real-time quantitative PCR(qPCR).Mouse peritoneal macrophages were infected with the H37Ra,ΔRv1140,and ΔRv1140+Rv1140 strains,and the transcriptional levels of inflammatory cytokines interleukin-1β(IL-1β),interleukin-6(IL-6),and tumor necrosis factor-α(TNF-α),as well as alongside intracellular colonization,were detected by qPCR.The results indicated both the ΔRv1140 strain and the ΔRv1140+Rv1140 strain were constructed successfully.Compared with the H37Ra strain,deletion of the Rv1140 protease did not affect H37Ra growth.Compared with that before infection,the mRNA relative expression level of the rv1140 gene significantly increased at 48 and 72 hours post infection in H37Ra-infected macrophages(P<0.001);in U937 cells,the mRNA relative expression of rv1140 was significantly or highly significantly upregulated at 24,48,and 72 hours post infection(P<0.05 or P<0.01 or P<0.001).Compared to ΔRv1140 strain infection,both H37Ra strain and the ΔRv1140+Rv1140 strain induced a highly significant increase in the mRNA relative expression of IL-1β at 12 h post infection(P<0.001).At 6 and 12 hours post infection,the mRNA relative expression levels of IL-6 and TNF-α were significantly elevated in cells infected with H37Ra and ΔRv1140+Rv1140(P<0.01 or P<0.001).These findings indicated that the Rv1140 protease not only participated in regulating the host immune response but also exhibited a negative correlation with the intracellular colonization ability of MTB in macrophages.关键词
结核分枝杆菌/蛋白酶/Rv1140/免疫反应/候选抗原Key words
Mycobacterium tuberculosis/protease/Rv1140/immunoreactive/candidate antigen分类
农业科技引用本文复制引用
郭若男,聂蝉蝉,崔莹莹,党光辉,刘思国,姜艳艳,陈春文,段祎凡,高云洁,张镇钧,冯婷婷,王慧,李梦雨..Rv1140蛋白酶在结核分枝杆菌感染中的作用研究[J].黑龙江畜牧兽医,2026,(1):21-29,9.基金项目
"十四五"国家重点研发计划项目(2021YFD1800403) （2021YFD1800403）
国家自然科学基金项目(32273005) （32273005）
中央级公益性科研院所基本科研业务费专项(1610302024002) （1610302024002）
