IN-VITRO AND IN-VIVO CHARACTERIZATION OF AN AIL MUTANT OF YERSINIA-ENTEROCOLITICA

Ail is a 17-kDa protein of Yersinia enterocolitica previously identified on the basis of its ability to confer upon Escherichia call the phenotype of attachment and invasion of cultured epithelial cells. Here we report an examination of the contribution of ail to the pathogenicity of Y. enterocolitica. A low copy-number ail plasmid that promoted serum resistance in E. coli HB101 was constructed. The serum resistance phenotype conferred by ail to E. coli was affected by the growth phase of the culture as well as by the gene copy number. In Contrast, the copy number of ail (and the relative quantity of Ail) was found to have little effect on the amount of Ail promoted invasion of cultured epithelial cells. An ail mutant of Y. enterocolitica was constructed and characterized in vitro. This mutant produced no detectable Ail and had a reduced ability to invade CHO cells. Serum resistance of Y. enterocolitica was Ail dependent and was affected by growth phase and ail copy number. The phenotype of the ail mutant was examined in vivo by using a murine model for infection. The ail mutant phenotype was identical to that of the wild-type strain in oral 50% lethal dose studies and early colonization of Peyer's patches as well as in kinetic studies. Western blot (immunoblot) analysis of Ail produced by bacteria growing in vivo at 48 h postinfection indicated that ail was expressed at this time point. Thus, our findings confirm that Ail contributes to the serum resistance and invasion phenotypes of Y. enterocolitica in vitro and indicate that Ail is not required to establish an infection or to cause systemic infection of BALB/c or DBA/2 mice.