Immunization with outer membrane proteins (OprF and OprI) and flagellin B protects mice from pulmonary infection with mucoid and nonmucoid Pseudomonas aeruginosa

J Microbiol Immunol Infect. 2018 Jun;51(3):312-320. doi: 10.1016/j.jmii.2016.08.014. Epub 2017 Feb 20.

Abstract

Background: Pseudomonas aeruginosa is a Gram-negative opportunistic bacterium, which considered as a common cause of nosocomial infection and life-threatening complications in immunocompromized and cystic fibrosis patients. Here, we evaluate the protective effect of recombinant vaccines composed of outer membrane proteins OprF and OprI alone or in combination with flagellin B against mucoid and nonmucoid pseudomonas infection.

Methods: BALB/C mice were immunized subcutaneous using OprF and OprI with or without flagellin B and antibody titers were determined. Serum bactericidal and opsonophagocytosis activities of immunized and control sera were estimated against mucoid and nonmucoid pseudomonas strains. Lung tissue sections from immunized and nonimmunized mice were analyzed and the levels of peripheral neutrophils infiltration into the lung and tissue inflammation were scored.

Results: Subcutaneous immunization using OprF and OprI with or without flagellin B elicited higher antibody titers against OprF, OprI, and flagellin B. The produced antibodies successfully opsonized both mucoid and nonmucoid strains with subsequent activation of the terminal pathway of complement that enhances killing of nonmucoid strains via complement-mediated lysis. Furthermore, opsonized mucoid and nonmucoid strains showed enhanced opsonophagocytosis via human peripheral neutrophils, a mechanism that kills P. aeruginosa when complement mediated lysis is not effective especially with mucoid strains. Immunized mice also showed a significant prolonged survival time, lower bacteremia, and reduced lung damage when compared with control nonimmunized mice.

Conclusion: Our data showed that mice immunized with OprF/OprI or OprF/OprI and flagellin B are significantly protected from infection caused by mucoid and nonmucoid strains of P. aeruginosa.

Keywords: Pseudomonas aeruginosa; flagellin B; mucoid; nonmucoid; outer membrane proteins.

MeSH terms

  • Animals
  • Antibodies, Bacterial / blood
  • Bacterial Outer Membrane Proteins / immunology*
  • Bacterial Proteins / genetics
  • Bacterial Proteins / immunology*
  • Cloning, Molecular
  • Disease Models, Animal
  • Female
  • Flagellin / genetics
  • Flagellin / immunology*
  • Gene Expression Regulation, Bacterial
  • Immunization*
  • Lipoproteins / genetics
  • Lipoproteins / immunology*
  • Lung / pathology
  • Mice
  • Mice, Inbred BALB C
  • Phagocytosis
  • Pseudomonas Infections / immunology
  • Pseudomonas Infections / microbiology
  • Pseudomonas Infections / prevention & control*
  • Pseudomonas Vaccines / genetics
  • Pseudomonas Vaccines / immunology*
  • Pseudomonas aeruginosa / genetics
  • Pseudomonas aeruginosa / immunology*
  • Pseudomonas aeruginosa / pathogenicity
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / immunology
  • Respiratory Tract Infections / microbiology
  • Respiratory Tract Infections / prevention & control*
  • Vaccination
  • Vaccines, Synthetic / genetics
  • Vaccines, Synthetic / immunology
  • Vaccines, Synthetic / therapeutic use

Substances

  • Antibodies, Bacterial
  • Bacterial Outer Membrane Proteins
  • Bacterial Proteins
  • Lipoproteins
  • OprF protein, Pseudomonas aeruginosa
  • Pseudomonas Vaccines
  • Recombinant Fusion Proteins
  • Vaccines, Synthetic
  • outer membrane lipoprotein I, Bacteria
  • Flagellin
  • flaB flagellin