Bengoechea J A, Díaz R, Moriyón I
Departamento de Microbiologia, Universidad de Navarra, Pamplona, Spain.
Infect Immun. 1996 Dec;64(12):4891-9. doi: 10.1128/iai.64.12.4891-4899.1996.
Sensitivities to polycationic peptides and EDTA were compared in Yersinia enterocolitica pathogenic and environmental biogroups. As shown by changes in permeability to the fluorescent hydrophobic probe N-phenylnaphthylamine (NPN), the outer membranes (OMs) of pathogenic and environmental strains grown at 26 degrees C in standard broth were more resistant to poly-L-lysine, poly-L-ornithine, melittin, cecropin P1, polymyxin B, and EDTA than Escherichia coli OMs. At 37 degrees C, OMs of pathogenic biogroups were resistant to EDTA and polycations and OMs of environmental strains were resistant to EDTA whereas E. coli OMs were sensitive to both EDTA and polycations. Similar results were found when testing deoxycholate sensitivity after polycation exposure or when isogenic pairs with or without virulence plasmid pYV were compared. With bacteria grown without Ca++ available, OM permeability to NPN was drastically increased in pathogenic but not in environmental strains or E. coli. Under these conditions, OMs of pYV+ and pYV- cells showed small differences in NPN permeability but differences in polycation sensitivity could not be detected by fluorimetry. O:1,6 (environmental type) lipopolysaccharide (LPS), but not O:3 or O:8 LPS, was markedly rough at 37 degrees C, and this could explain the differences in polycation sensitivity. LPSs from serotypes O:3 and O:8 grown at 37 degrees C were more permeable to NPN than O:1,6 LPS, and O:8 LPS was resistant to polycation-induced permeabilization. These data suggest that LPSs relate to some but not all the OM differences described. It is hypothesized that the different OM properties of environmental and pathogenic biogroups reflect the adaptation of the latter biogroups to pathogenicity.
比较了小肠结肠炎耶尔森氏菌致病生物群和环境生物群对聚阳离子肽和乙二胺四乙酸(EDTA)的敏感性。如对荧光疏水探针N-苯基萘胺(NPN)的通透性变化所示,在标准肉汤中于26℃培养的致病菌株和环境菌株的外膜,比大肠杆菌的外膜对聚-L-赖氨酸、聚-L-鸟氨酸、蜂毒素、天蚕素P1、多粘菌素B和EDTA更具抗性。在37℃时,致病生物群的外膜对EDTA和聚阳离子具有抗性,环境菌株的外膜对EDTA具有抗性,而大肠杆菌的外膜对EDTA和聚阳离子均敏感。在聚阳离子暴露后测试脱氧胆酸盐敏感性时,或比较有无毒力质粒pYV的同基因对时,发现了类似结果。在没有Ca++的情况下培养细菌时,致病菌株中外膜对NPN的通透性急剧增加,而环境菌株或大肠杆菌中则没有。在这些条件下,pYV+和pYV-细胞的外膜在NPN通透性上显示出微小差异,但荧光法无法检测到聚阳离子敏感性的差异。O:1,6(环境型)脂多糖(LPS),而非O:3或O:8 LPS,在37℃时明显粗糙,这可以解释聚阳离子敏感性的差异。在37℃培养的O:3和O:8血清型的LPS比O:1,6 LPS对NPN的通透性更高,并且O:8 LPS对聚阳离子诱导的通透化具有抗性。这些数据表明,LPS与所描述的部分而非全部外膜差异有关。据推测,环境生物群和致病生物群不同的外膜特性反映了后者对致病性的适应性。