Opa+ Neisseria Gonorrhoeae Exhibits Reduced Survival in Human Neutrophils via Src Family Kinase-Mediated Bacterial Trafficking Into Mature Phagolysosomes

Cell Microbiol. 2015 May;17(5):648-65. doi: 10.1111/cmi.12389. Epub 2014 Nov 25.

Abstract

During gonorrhoeal infection, there is a heterogeneous population of Neisseria gonorrhoeae (Gc) varied in their expression of opacity-associated (Opa) proteins. While Opa proteins are important for bacterial attachment and invasion of epithelial cells, Opa+ Gc has a survival defect after exposure to neutrophils. Here, we use constitutively Opa- and OpaD+ Gc in strain background FA1090 to show that Opa+ Gc is more sensitive to killing inside adherent, chemokine-treated primary human neutrophils due to increased bacterial residence in mature, degradative phagolysosomes that contain primary and secondary granule antimicrobial contents. Although Opa+ Gc stimulates a potent oxidative burst, neutrophil killing of Opa+ Gc was instead attributable to non-oxidative components, particularly neutrophil proteases and the bactericidal/permeability-increasing protein. Blocking interaction of Opa+ Gc with carcinoembryonic antigen-related cell adhesion molecules (CEACAMs) or inhibiting Src family kinase signalling, which is downstream of CEACAM activation, enhanced the survival of Opa+ Gc in neutrophils. Src family kinase signalling was required for fusion of Gc phagosomes with primary granules to generate mature phagolysosomes. Conversely, ectopic activation of Src family kinases or coinfection with Opa+ Gc resulted in decreased survival of Opa- Gc in neutrophils. From these results, we conclude that Opa protein expression is an important modulator of Gc survival characteristics in neutrophils by influencing phagosome dynamics and thus bacterial exposure to neutrophils' full antimicrobial arsenal.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacterial Outer Membrane Proteins / metabolism*
  • Cell Adhesion Molecules / metabolism
  • Cells, Cultured
  • Humans
  • Microbial Viability*
  • Neisseria gonorrhoeae / immunology*
  • Neisseria gonorrhoeae / physiology*
  • Neutrophils / immunology*
  • Neutrophils / microbiology*
  • Phagosomes / microbiology*
  • Signal Transduction

Substances

  • Bacterial Outer Membrane Proteins
  • Cell Adhesion Molecules
  • Opa protein, Neisseria