Objectives: Pseudomonas aeruginosa infections are associated with considerable morbidity and mortality in critically ill patients because of multidrug resistance. Our objective in this study was to determine the protective effects of murine monoclonal antibodies (MAbs) against a fusion protein containing PilQ and disulphide turn region of PilA (QA) from P. aeruginosa in a model of burn wound sepsis. Methods: After expression in E. coli BL21 strain, recombinant proteins were adsorbed to Ni-NTA, and confirmed by Western blotting The purified recombinant protein was used as an antigen to immunize mice. Immunized murine spleen cells were fused with myeloma cells and hybridoma cells were cultured in HAT medium. A number of antibody-producing hybridomas were cloned by limiting dilution method. Then, for mass production of MAbs, ascites fluid formation method was used in mice and purified by affinity chromatography. Final confirmation of MAbs purity was performed by SDS-PAGE method. Twitching motility inhibition assay was performed to evaluate the biological activity of the produced MAbs. After modeling the burn in mice, P. aeruginosa O1 infection was induced by injecting bacteria with a concentration of 3 times LD50 (3-5 × 102 cfu / ml) in the burned area and mortality was assessed in mice treated with MAbs compared with control groups for 7 days. Results: The 1H10-D6 and 2E1-E6 MAbs produced against the PilQ-PilA DSL region sequence completely inhibited the motility of the PAO1 strain. Also, the results showed that mice receiving 2E1-E6 provided significant protection against bacterial strains. Reliably, in comparison with other treatment groups, the combination of antibiotic and 2E1-E6 mAb essentially enhanced the survival of mice infected with PAO1. Conclusion: Consequently, the antibiotic and 2E1-E6 MAb combination gives a new effective strategy for the treatment of P. aeruginosa sepsis, particularly when large numbers of exceptionally virulent strains are present.
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