title: Protective efficacy of recombinant exotoxin A--flagellin fusion protein against Pseudomonas aeruginosa infection.
abstract: Pseudomonas aeruginosa is an opportunistic bacterium that causes serious nosocomial infection in immunocompromised patients. The aim of this study was to prepare a fusion protein consisting of exotoxin A (ExoA) and flagellin (Fla) from P. aeruginosa and to evaluate its potential as a vaccine candidate against P. aeruginosa infection. The genes encoding for ExoA and Fla proteins were cloned in-frame and expressed in Escherichia coli. The recombinant ExoA-Fla fusion protein was purified by Ni-NTA affinity chromatography. Mice were immunized subcutaneously with ExoA, Fla, and ExoA-Fla fusion proteins, and the humoral immune response was evaluated by ELISA method. The immunized and control group mice were challenged with a 2× LD50 (7.5 × 10(7) CFU) of P. aeruginosa for the protection assay. The results indicated that vaccination with Fla, ExoA, and ExoA-Fla fusion proteins produced a significant amount of specific immunoglobulin G antibodies. Immunization of mice with ExoA-Fla fusion protein showed significant protection against intraperitoneal challenge with 7.5 × 10(7) CFU (2× LD50) P. aeruginosa. Results of this study suggest that recombinant ExoA-Fla fusion protein is a highly immunogenic protective protein showing promise as a vaccine candidate against P. aeruginosa.
authors.affiliation: a Research Center for Infectious and Tropical Disease, Tabriz University of Medical Sciences, Tabriz, Iran.
authors.forename: Safar
authors.initials: S
authors.lastname: Farajnia
authors.forename: Shahin Najar
authors.initials: SN
authors.lastname: Peerayeh
authors.forename: Asghar
authors.initials: A
authors.lastname: Tanomand
authors.forename: Jafar
authors.initials: J
authors.lastname: Majidi
authors.forename: Gholamreza
authors.initials: G
authors.lastname: Goudarzi
authors.forename: Behrooz
authors.initials: B
authors.lastname: Naghili
authors.forename: Leila
authors.initials: L
authors.lastname: Rahbarnia
mesh_terms: D000818:Animals; D000907:Antibodies, Bacterial / Q000276:immunology; D001426:Bacterial Proteins / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology*; D001428:Bacterial Vaccines / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology*; D005098:Exotoxins / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology*; D005260:Female; D005408:Flagellin / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology*; D006801:Humans; D056724:Immunity, Humoral; D007114:Immunization; D051379:Mice; D008807:Mice, Inbred BALB C; D011552:Pseudomonas Infections / Q000276:immunology / Q000382:microbiology / Q000517:prevention & control*; D011550:Pseudomonas aeruginosa / Q000235:genetics / Q000276:immunology* / Q000502:physiology; D011994:Recombinant Proteins / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology; D014611:Vaccination
chemical_list: D000907:Antibodies, Bacterial; D001426:Bacterial Proteins; D001428:Bacterial Vaccines; D005098:Exotoxins; D011994:Recombinant Proteins; D005408:Flagellin
keywords: Pseudomonas aeruginosa; candidat vaccin; exotoxin A – flagellin; exotoxine A – flagelline; fusion protein; protéine de fusion; vaccine candidate