Banemann A, Deppisch H, Gross R
Lehrstuhl für Mikrobiologie, Theodor-Boveri-Institut, Biozentrum der Universität Würzburg, D-97074 Würzburg, Germany.
Infect Immun. 1998 Dec;66(12):5607-12. doi: 10.1128/IAI.66.12.5607-5612.1998.
Resistance profiles of the two Bordetella species B. bronchiseptica and B. pertussis against various antimicrobial peptides were determined in liquid survival and agar diffusion assays. B. bronchiseptica exhibited significantly higher resistance against all tested peptides than B. pertussis. The most powerful agents acting on B. bronchiseptica were, in the order of their killing efficiencies, cecropin P > cecropin B > magainin-II-amide > protamine > melittin. Interestingly, for B. bronchiseptica, the resistance level was significantly affected by phase variation, as a bvgS deletion derivative showed an increased sensitivity to these peptides. Tn5-induced protamine-sensitive B. bronchiseptica mutants, which were found to be very susceptible to most of the cationic peptides, were isolated. In two of these mutants, the genetic loci inactivated by transposon insertion were identified as containing genes highly homologous to the wlbA and wlbL genes of B. pertussis that are involved in the biosynthesis of lipopolysaccharide (LPS). In agreement with this finding, the two peptide-sensitive mutants revealed structural changes in the LPS, resulting in the loss of the O-specific side chains and the prevalence of the LPS core structure. This demonstrates that LPS plays a major role in the resistance of B. bronchiseptica against the action of antimicrobial peptides and suggests that B. pertussis is much more susceptible to these peptides due to the lack of the highly charged O-specific sugar side chains.
通过液体存活试验和琼脂扩散试验，测定了支气管败血博德特氏菌和百日咳博德特氏菌这两种博德特氏菌对各种抗菌肽的抗性谱。支气管败血博德特氏菌对所有测试肽的抗性均显著高于百日咳博德特氏菌。按照杀伤效率排序，作用于支气管败血博德特氏菌的最有效药物依次为：天蚕素P＞天蚕素B＞蛙皮素-II-酰胺＞鱼精蛋白＞蜂毒肽。有趣的是，对于支气管败血博德特氏菌，抗性水平受相变显著影响，因为bvgS缺失衍生物对这些肽的敏感性增加。分离出了Tn5诱导的对鱼精蛋白敏感的支气管败血博德特氏菌突变体，发现它们对大多数阳离子肽非常敏感。在其中两个突变体中，被转座子插入失活的基因位点被鉴定为含有与百日咳博德特氏菌参与脂多糖（LPS）生物合成的wlbA和wlbL基因高度同源的基因。与此发现一致，这两个对肽敏感的突变体显示出LPS的结构变化，导致O-特异性侧链缺失，LPS核心结构占优势。这表明LPS在支气管败血博德特氏菌对抗菌肽作用的抗性中起主要作用，并表明百日咳博德特氏菌由于缺乏高度带电的O-特异性糖侧链而对这些肽更敏感。
