Nassif X, Fournier J M, Arondel J, Sansonetti P J
Service des Entérobactéries, Institut National de la Santé et de la Recherche Médicale, Paris, France.
Infect Immun. 1989 Feb;57(2):546-52. doi: 10.1128/iai.57.2.546-552.1989.
We have previously reported that the presence of a 180-kilobase plasmid encoding production of aerobactin was correlated with the virulence of Klebsiella pneumoniae K1 and K2 isolates. This work demonstrates that a variant of a K2 strain which has lost this plasmid, pKP100, becomes avirulent. Labeling of this plasmid with the mobilizable, replication-defective element pME28, used here as a mobilizable transposon, allowed the transfer of this plasmid into a plasmidless derivative. Virulence was restored upon reacquisition of this tagged plasmid, pKP101. In addition to aerobactin production, another phenotype could be correlated with the presence of this virulence plasmid: the mucoid phenotype of the bacterial colonies. Both wild-type and plasmidless strains are encapsulated, but only the former presented mucoid colonies. Participation of this phenotype in the virulence of K. pneumoniae was demonstrated by constructing a mutant altered in the plasmid gene encoding this phenotype. The resulting strain demonstrated a 1,000-fold decrease in virulence. Introduction of the recombinant plasmid pKP200 carrying the gene encoding this mucoid phenotype into Escherichia coli HB101 also led to the production of a mucoid phenotype. Rocket immunoelectrophoresis demonstrated that in E. coli this phenotype was due to the production of colanic acid. On the other hand, neither the overproduction of K2 capsular polysaccharide nor the presence of colanic acid was detected in mucoid strains of K. pneumoniae. We conclude that this mucoid phenotype is definitely an important virulence factor of K. pneumoniae. It is due to the plasmid-encoded production of a substance which is different from colanic acid and the capsular polysaccharide of K. pneumoniae.
我们之前曾报道,编码气杆菌素产生的180千碱基质粒的存在与肺炎克雷伯菌K1和K2分离株的毒力相关。这项研究表明,丢失该质粒(pKP100)的K2菌株变体变得无毒。用可移动的、复制缺陷元件pME28标记该质粒,此处用作可移动转座子,可使该质粒转移到无质粒衍生物中。重新获得这种标记质粒pKP101后,毒力得以恢复。除了产生气杆菌素外,另一种表型也可能与这种毒力质粒的存在相关:细菌菌落的黏液样表型。野生型和无质粒菌株都有荚膜,但只有前者呈现黏液样菌落。通过构建编码该表型的质粒基因发生改变的突变体,证明了该表型在肺炎克雷伯菌毒力中的作用。所得菌株的毒力降低了1000倍。将携带编码这种黏液样表型基因的重组质粒pKP200导入大肠杆菌HB101也导致了黏液样表型的产生。火箭免疫电泳表明,在大肠杆菌中,这种表型是由于产生了柯氏酸。另一方面,在肺炎克雷伯菌的黏液样菌株中未检测到K2荚膜多糖的过量产生或柯氏酸的存在。我们得出结论,这种黏液样表型肯定是肺炎克雷伯菌的一种重要毒力因子。它是由于质粒编码产生了一种不同于肺炎克雷伯菌的柯氏酸和荚膜多糖的物质。