Manterola Lorea, Guzmán-Verri Caterina, Chaves-Olarte Esteban, Barquero-Calvo Elías, de Miguel María-Jesús, Moriyón Ignacio, Grilló María-Jesús, López-Goñi Ignacio, Moreno Edgardo
Departamento de Microbiología y Parasitología, Universidad de Navarra, 31008 Pamplona, Spain.
Infect Immun. 2007 Oct;75(10):4867-74. doi: 10.1128/IAI.00439-07. Epub 2007 Jul 30.
The Brucella abortus two-component regulatory system BvrR/BvrS controls the expression of outer membrane proteins (Omp) Omp3a (Omp25) and Omp3b (Omp22). Disruption of bvrS or bvrR generates avirulent mutants with altered cell permeability, higher sensitivity to microbicidal peptides, and complement. Consequently, the role of Omp3a and Omp3b in virulence was examined. Similar to bvrS or bvrR mutants, omp3a and omp3b mutants displayed increased attachment to cells, indicating surface alterations. However, they showed unaltered permeability; normal expression of Omp10, Omp16, Omp19, Omp2b, and Omp1; native hapten polysaccharide; and lipopolysaccharide and were resistant to complement and polymyxin B at ranges similar to those of the wild-type (WT) counterpart. Likewise, omp3a and omp3b mutants were able to replicate in murine macrophages and in HeLa cells, were resistant to the killing action of human neutrophils, and persisted in mice, like the WT strain. Murine macrophages infected with the omp3a mutant generated slightly higher levels of tumor necrosis factor alpha than the WT, whereas the bvrS mutant induced lower levels of this cytokine. Since the absence of Omp3a or Omp3b does not result in attenuation, it can be concluded that BvrR/BvrS influences additional Brucella properties involved in virulence. Our results are discussed in the light of previous works suggesting that disruption of omp3a generates attenuated Brucella strains, and we speculate on the role of group 3 Omps.
流产布鲁氏菌双组分调节系统BvrR/BvrS控制外膜蛋白(Omp)Omp3a(Omp25)和Omp3b(Omp22)的表达。bvrS或bvrR的破坏会产生无毒力突变体,这些突变体具有改变的细胞通透性、对杀菌肽更高的敏感性以及补体敏感性。因此,研究了Omp3a和Omp3b在毒力中的作用。与bvrS或bvrR突变体相似,omp3a和omp3b突变体对细胞的附着增加,表明表面发生了改变。然而,它们的通透性未改变;Omp10、Omp16、Omp19、Omp2b和Omp1表达正常;天然半抗原多糖以及脂多糖正常,并且在与野生型(WT)相当的范围内对补体和多粘菌素B具有抗性。同样,omp3a和omp3b突变体能够在小鼠巨噬细胞和HeLa细胞中复制,对人类中性粒细胞的杀伤作用具有抗性,并且像WT菌株一样在小鼠体内持续存在。感染omp3a突变体的小鼠巨噬细胞产生的肿瘤坏死因子α水平略高于WT,而bvrS突变体诱导的这种细胞因子水平较低。由于缺乏Omp3a或Omp3b不会导致毒力减弱,因此可以得出结论,BvrR/BvrS会影响布鲁氏菌其他与毒力相关的特性。我们根据之前的研究结果进行了讨论,之前的研究表明omp3a的破坏会产生减毒的布鲁氏菌菌株,并且我们推测了第3组Omp的作用。