An approach to chimeric subunit immunogen provides efficient protection against toxicity, type III and type v secretion systems of Shigella

Abstract

ObjectivesShigellosis is one of the infectious diseases causing severe intestinal illness in human beings. Development of an effective vaccine against Shigella is a key to deal with this bacterium. The present study aimed at evaluation of the antibody response as well as the protection of the recombinant chimeric protein containing IpaD, IpaB, StxB, and VirG against Shigella dysentery and flexneri. MethodsChimeric protein was expressed and purified by Ni-NTA resin. The identity of the protein was determined by Western blot analysis. Mouse groups were immunized with the recombinant protein and the humoral immune response was measured by Enzyme-Linked Immunosorbent Assay (ELISA). Additionally, neutralization of the bacterial toxin by antibody was assessed by MTT assay. Animal challenge against S.dysentery and S. flexneri was evaluated, as well. ResultsProtein expression and purification were confirmed by SDS-PAGE and western blotting. Analysis of the immune responses demonstrated that the antibody responses were higher in the sera of the subcutaneously immunized mice compared to those immunized intraperitoneally. In vitro neutralization analysis indicated that the 1:10000 dilution of the sera had a high ability to neutralize 0.25 ng/µl (CD50) of the toxin on the Vero cell line. Furthermore, the results of the animal challenge showed that the immunized mice were completely protected against 50 LD50 of the bacterial toxin. Immunization also protected 80% of the mice from 10 LD50 by S. flexneri and S.dysentery. In addition, passive immunization conferred 60% protection in the mice against S. flexneri and S.dysentery. Organ burden studies also revealed a significant reduction in infection among the immunized mice. ConclusionThis study revealed that the chimeric protein produced inE. colicould be a promising chimeric immunogen candidate against Shigella.