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Immunization against Vibrio cholerae, ETEC, and EHEC with chitosan nanoparticle containing LSC chimeric protein

Ghaffari Marandi, Bentol Hoda, Zolfaghari, Mohammad Reza, Kazemi, Rouhollah, Motamedi, Mohammad Javad, Amani, Jafar
Microbial pathogenesis 2019 v.134 pp. 103600
Vibrio cholerae, Western blotting, antibodies, antigens, bacteria, chitosan, developing countries, diarrhea, encapsulation, enterohemorrhagic Escherichia coli, enterotoxigenic Escherichia coli, enzyme-linked immunosorbent assay, genes, immunization, immunoglobulin A, immunoglobulin G, intestines, mice, mortality, mucosal immunity, nanoparticles, neutralization, protective effect, recombinant fusion proteins, toxins, vaccines
Severe intestinal infections caused by V. cholerae, ETEC and EHEC have contributed to the mortality rate in developing countries. Vibrio Cholera, ETEC and EHEC bacterium with the production of CT, LT and Stx2 toxins respectively lead to severe watery and bloody diarrhea. This study aimed to investigate a trimeric vaccine candidate containing recombinant chimeric protein, encapsulate the protein in chitosan nanoparticles and assess its immunogenicity.The LSC recombinant gene was used. It is composed of LTB (L), STXB (S) and CTXB (C) subunits respectively. The LSC recombinant protein was expressed and purified and confirmed by western blotting. The purified protein was encapsulated in chitosan nanoparticles, and its size was measured. BalB/c mice were immunized in four groups through oral and injection methods by LSC protein. The antibody titer was then evaluated by ELISA, and finally, the challenge test of the toxins from all three bacteria was done on the immunized mouse.After expression and purification LSC protein size of nanoparticles containing protein was measured at 104.6 nm. Nanoparticles were able to induce systemic and mucosal immune responses by generating a useful titer of IgG and IgA. The challenge results with LT, CT and Stx toxins showed that the LSC protein might partially neutralize the effect of toxins.LSC chimeric protein with the simultaneous three essential antigens have a protective effect against the toxins produced by ETEC, EHEC and Vibrio cholera bacteria and it can be used in vaccines to prevent Diarrhea caused by these three bacteria.