Yersinia enterocolitica O:9 as a potential live oral carrier for protective antigens

Yersinia enterocolitica has the capacity to invade the intestinal tissue and to resist the primary host resistance. The former is chromosome coded while the second largely depends on the presence of a 70 kb plasmid called pYV. This plasmid directs the conditional synthesis of high amounts of proteins (YOPs) that are secreted and inserted in the outer membrane. In order to evaluate Y. enterocolitica W22703 as a potential live carrier for immunization, three strains expressing beta-galactosidase (GZ), were tested for their ability to induce an antibody response to this antigen in mice. The first strain contained plasmid pGC1256, a mutated pYV plasmid containing lacZ transcribed from a yop gene promoter. This strain produced high amounts of GZ instead of a YOP protein and was shown to be hypovirulent. The other strains tested were W22703 pYV+ and pYV- containing a derepressed lac operon carried on an independent plasmid. Immunoblot analysis of sera of mice having received by oral inoculation, W22703(pGC1256) or the pYV+ GZ producing strain revealed the presence of antibodies to GZ. The response to GZ after inoculation of W22703(pGC1256) was shown by ELISA to be only slightly inferior to that obtained by subcutaneous injection of GZ. No response was obtained after oral inoculation of the pYV-GZ producing strain. This showed that the presence of pYV was necessary to obtain an antibody response in this system.