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Inhibition of H. pylori colonization and prevention of gastritis in murine model

Ameri Shah Reza, Mahdieh, Mousavi Gargari, Seyed Latif, Rasooli, Iraj, Jalali Nadoushan, Mohammadreza, Ebrahimizadeh, Walead
World journal of microbiology & biotechnology 2012 v.28 no.7 pp. 2513-2519
Gram-negative bacteria, Helicobacter pylori, Western blotting, affinity chromatography, animal disease models, antibodies, blood sampling, enzyme-linked immunosorbent assay, gastric mucosa, gastritis, immune response, immunization, laboratory animals, mice, polyacrylamide gel electrophoresis, recombinant proteins, urease
Helicobacter pylori is a Gram-negative spiral bacterium that colonizes human gastric mucosa causing infection. In this study aiming at inhibition of H. pylori infection we made an attempt to evaluate immunogenicity of the total (UreC) and C-terminal (UreCc) fragments of H. pylori urease. Total UreC and its C-terminal fragment were expressed in E. coli. Recombinant proteins were analyzed by SDS-PAGE and western blot and then purified by Ni–NTA affinity chromatography. Female C57BL6/j mice were immunized with the purified proteins (UreC and UreCc). Antibody titers from isolated sera were measured by ELISA. Immunized mice were then challenged by oral gavage with live H. pylori Sydney strain SS1. Total of 109 CFU were inoculated into stomach of immunized and unimmunized healthy mice three times each at one day interval. Eight weeks after the last inoculation, the blood sample was collected and the serum antibody titer was estimated by ELISA. Stomach tissues from control and experimental animal groups were studied histopathologically. UreC and UreCc yielded recombinant proteins of 61 and 31 kDa respectively. ELIZA confirmed establishment of immunity and the antibodies produced thereby efficiently recognized H. pylori and inhibited its colonization in vivo. Pathological analysis did not reveal established infection in immunized mice challenged with H. pylori. The results support the idea that UreC and UreCc specific antibodies contribute to protection against H. pylori infections.