Document Type : Original Article
Authors
1 Department of Microbiology, Karaj Branch, Islamic Azad University, Karaj, Iran.
2 Departement of Microbiology, Maragheh University of Medical sciences, Maragheh, Iran.
3 Department of Immunology and Immunology Research Center, Faculty of Medicine, Tabriz University of Medical Sciences, Tabriz, Iran.
Abstract
Background: Pseudomonas aeruginosa is an important opportunistic pathogen, especially in patients with compromised host defense. Objective: To prepare the conjugate of detoxified lipopolysaccharide (D-LPS) and exotoxin A toxoid (T-ETA) from P. aeruginosa in gold nanoparticles (Au NPs) in a mice model. Methods: LPS and ETA were purified from P. aeruginosa PAO1. D-LPS was conjugated with T-ETA via the amidation method. Au NPs were bound to D-LPS-T-ETA conjugate via electrostatic interaction. Mice were immunized with D-LPS, D-LPS-Au NPs, T-ETA, T-ETA-Au NPs, D-LPS-T-ETA, D-LPS-T-ETA-Au NPs, D-LPS-Au NPs+T-ETA-Au NPs, Au NPs, and phosphate-buffered saline (PBS), and specific IgG titers were determined by the ELISA and the whole-cell ELISA methods. Mice in the vaccinated and control groups were exposed to a 2×LD50 of P. aeruginosa and mortality rates were recorded for one week. Results: The results showed that vaccination by D-LPS, D-LPS-Au NPs, T-ETA, T-ETA-Au NPs, D-LPS-T-ETA, D-LPS-T-ETA-Au NPs and D-LPS-Au NPs+T-ETA-Au NPs induced specific IgG. Mice received the D-LPS-T-ETA-Au NPs conjugate showed significant protection against bacterial challenge. Conclusion: These data indicate that D-LPS-T-ETA-Au NPs conjugate has a significant immunogenicity potential to be applied as a new vaccine against Pseudomonas infections.
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