Immunization with outer membrane vesicles of avian pathogenic Escherichia coli O78 induces protective immunity in chickens

Avian pathogenic Escherichia coli (APEC) typically causes colibacillosis and is a major concern for the poultry industry and public health. As a vaccine platform, the outer membrane vesicles (OMVs) derived from various gram-negative bacteria and even some gram-positive bacteria have been reported to be immunogenic in laboratories or upon commercial usage worldwide. Here, we purified OMVs from APEC serotype 078 strain by ultracentrifugation and gradient isolation. By SDS-PAGE and LC-MS/MS analysis, the 20 most abundant proteins located on OMVs were identified and analyzed; the lipopolysaccharide (LPS) profiles of OMVs were not different from those of the bacteria. Moreover, three groups of chickens were immunized with OMV-, outer membrane protein (OMP)- and PBS, with the latter two serving as positive and negative controls, respectively. By analyzing the anti-OMP and anti-LPS IgG titers stimulated by the tested vaccine candidates, the macrophage opsonophagocytic activity and the bactericidal activity mediated by serum antibodies in vaccinated chickens, we found that the OMV-vaccinated chicken group was superior to the two other groups. These findings were confirmed by additional chicken challenge tests, in which all OMV-vaccinated group chickens obtained complete protection but those of the other two groups were barely protected. Our data demonstrate that native APEC 078 OMVs can induce protective immunity in chickens and therefore be used as a candidate vaccine for APEC serotype 078 strain infections.