Yersinia enterocolitica, a primary model for bacterial invasiveness.

Yersinia enterocolitica is now the species of Yersinia most frequently isolated from human and animal infections. The species includes pathogens and ubiquitous strains. Among the human pathogens, those isolated in America are more virulent than those isolated elsewhere, especially in Europe and Japan, and these isolates differ biochemically and serologically. The relation between Y. enterocolitica and Y. pestis only became obvious in 1980 with the discovery that at 37 degrees C Y. enterocolitica requires Ca++, a phenotype described in the 1960s for Y. pestis. This requirement as well as virulence is dependent on a 70-kilobase plasmid found later in Y. pseudotuberculosis and Y. pestis. Thus, many bacteriologists elected Y. enterocolitica as a model for bacterial invasiveness. However, studies with non-American strains were impeded by the lack of an inexpensive, simple animal test, a difficulty now circumvented by supplying an appropriate siderophore to the bacteria. Ca++ dependence can be viewed as a transition between free growth and protection against the immune system. In the latter phase, Y. enterocolitica synthesizes and releases large amounts of six plasmid-encoded outer membrane proteins. Most of these are under the control of the plasmid region governing Ca++ dependence. Mutants in this region either lose the Ca++ requirement at 37 degrees C or become unable to grow at 37 degrees C irrespective of the Ca++ concentration. The complex events leading to Ca++ dependence is still not understood. Virulence in Y. enterocolitica also depends on chromosomal genes: the endocytosis in intestinal epithelial cells seems not to be encoded by the pYV plasmid. Studies of Y. pseudotuberculosis suggest that this property depends on a single chromosomal locus, the study of which might be particularly important in the understanding of the first step in infection.

Genetic analysis of virulence plasmid from a serogroup 9 Yersinia enterocolitica strain: role of outer membrane protein P1 in resistance to human serum and autoagglutination.

Enteropathogenic strains of Yersinia enterocolitica harbor a virulence plasmid (70 kilobases) which specifies, at 37 degrees C, a calcium requirement for growth, autoagglutinability, resistance to the bactericidal activity of human serum, and the expression of some outer membrane proteins (OMPs). To map the genes encoding these properties, the virulence plasmid of a serogroup 9 strain (W22708) was subjected to transposon mutagenesis. A set of 68 independent mutations was obtained in Escherichia coli by transposon Tn813 (a tnpR mutant of Tn21)-mediated cointegration with the self-transmissible R388 plasmid. The resulting cointegrates were introduced and studied in Y. enterocolitica W22708. One mutant lost the calcium dependence property. Two other mutants presented a peculiar phenotype: they grew poorly at 37 degrees C, especially in the presence of calcium. Lastly, two mutants were affected in the properties of autoagglutination and resistance to human serum. Analysis of the OMP pattern of these two mutants revealed the absence of the largest OMP, called P1 (I. Bölin, and H. Wolf-Watz, Infect. Immun. 43:72-78, 1984). Complementation of one of these mutations with the cloned structural gene of OMP P1 restored the wild-type phenotype. However, OMP P1 was not sufficient by itself to specify the serum resistance property and a rapid autoagglutination of the host.

A simple adult-mouse test for tissue invasiveness in Yersinia enterocolitica strains of low experimental virulence.

The virulence of Yersinia enterocolitica depends on the presence of a 70-kilobase plasmid, called the Vwa plasmid. This situation is particularly favourable for studies of the mechanism of pathogenicity, but these are hindered by the lack of a suitable animal test to monitor the virulence of the human-pathogenic strains isolated outside the USA which belong to serogroups O:3, O:9 and O:5,27. We observed that, after oral administration to the mouse, the Vwa-positive strains of these serogroups produce a discrete systemic infection while the Vwa-negative strains do not. We present here a simple mouse-virulence test based on this observation.