Omp16, a conserved peptidoglycan-associated lipoprotein, is involved in Brucella virulence in vitro
Omp16, a conserved peptidoglycan-associated lipoprotein, is involved in Brucella virulence in vitro
复制标题
DOI：
10.1007/s12275-020-0144-y
复制
发表时间：
2020-09-01
影响因子：
3
通讯作者：
Wang, Aihua
中科院分区：
文献类型：
--
作者：
Zhi, Feijie;Zhou, Dong;Wang, Aihua
布鲁氏菌是常见的人畜共患布鲁氏菌病的病原体，主要感染特定的动物物种。布鲁氏菌外膜蛋白（Omps）与布鲁氏菌光滑株的毒力有关，在疫苗研制和诊断试验中具有重要的应用价值。Omp 16是肽聚糖相关脂蛋白（帕尔斯）的同源物，迄今为止尚未在任何布鲁氏菌菌株中产生Omp 16突变体。Omp16基因在布鲁氏菌中的功能和致病机制尚不清楚，本研究证实Omp16基因在布鲁氏菌中具有高度保守的结构域，并尝试将Omp16基因在猪布鲁菌疫苗株2（B.suisS2）中缺失，但未成功，这表明Omp16基因对布鲁氏菌的生存至关重要。我们收购了AB。suisS2 Omp16突变体。Omp 16缺陷可使体外培养的布鲁氏菌外膜完整性和活性受损，Omp 16的失活可降低巨噬细胞RAW 264.7细胞内细菌的存活率。suisS2及其衍生物诱导Raw 264.7细胞中IL-1 β、IL-6和TNF-α mRNA的显著表达。而布鲁氏菌Omp16的失活则增强了Raw 264.7细胞中IL-1 β和IL-6的表达。这些结果表明，布鲁氏菌Omp 16突变体是通过条件互补获得的，并证实了Omp 16在体外和体内均能维持外膜完整性，并参与布鲁氏菌的毒力，这些结果对揭示布鲁氏菌的致病机制具有重要意义。
Brucella, the bacterial agent of common zoonotic brucellosis, primarily infects specific animal species. TheBrucellaouter membrane proteins (Omps) are particularly attractive for developing vaccine and improving diagnostic tests and are associated with the virulence of smoothBrucellastrains. Omp16 is a homologue to peptidoglycan-associated lipoproteins (Pals), and anomp16mutant has not been generated in anyBrucellastrain until now. Very little is known about the functions and pathogenic mechanisms of Omp16 inBrucella.Here, we confirmed that Omp16 has a conserved Pal domain and is highly conserved inBrucella.We attempted to deleteomp16inBrucella suisvaccine strain 2 (B.suisS2) without success, which shows that Omp16 is vital forBrucellasurvival. We acquired aB. suisS2 Omp16 mutant via conditional complementation. Omp16 deficiency impairedBrucellaouter membrane integrity and activityin vitro.Moreover, inactivation of Omp16 decreased bacterial intracellular survival in macrophage RAW 264.7 cells.B. suisS2 and its derivatives induced marked expression of IL-1 beta, IL-6, and TNF-a mRNA in Raw 264.7 cells. Whereas inactivation of Omp16 inBrucellaenhanced IL-1 beta and IL-6 expression in Raw 264.7 cells. Altogether, these findings show that theBrucellaOmp16 mutant was obtained via conditional complementation and confirmed that Omp16 can maintain outer membrane integrity and be involved in bacterial virulence inBrucella in vitroandin vivo.These results will be important in uncovering the pathogenic mechanisms ofBrucella.