Characterization and pathogenic role of outer membrane vesicles produced by the fish pathogen Piscirickettsia salmonis under in vitro conditions

Vet Microbiol. 2016 Feb 29:184:94-101. doi: 10.1016/j.vetmic.2015.09.012. Epub 2015 Nov 7.

Abstract

Piscirickettsia salmonis is one of the major fish pathogens affecting Chilean aquaculture. This Gram-negative bacterium is highly infectious and is the etiological agent of Piscirickettsiosis. Little is currently known about how the virulence factors expressed by P. salmonis are delivered to host cells. However, it is known that several Gram-negative microorganisms constitutively release outer membrane vesicles (OMVs), which have been implicated in the delivery of virulence factors to host cells. In this study, OMVs production by P. salmonis was observed during infection in CHSE-214 cells and during normal growth in liquid media. The OMVs were spherical vesicles ranging in size between 25 and 145 nm. SDS-PAGE analysis demonstrated that the protein profile of the OMVs was similar to the outer membrane protein profile of P. salmonis. Importantly, the bacterial chaperonin Hsp60 was found in the OMVs of P. salmonis by Western-blot and LC-MS/MS analyses. Finally, in vitro infection assays showed that purified OMVs generated a cytopathic effect on CHSE-214 cells, suggesting a role in pathogenesis. Therefore, OMVs might be an important vehicle for delivering effector molecules to host cells during P. salmonis infection.

Keywords: Hsp60; Outer membrane vesicles (OMV); Piscirickettsia salmonis; Piscirickettsiosis; Virulence.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Bacterial Outer Membrane Proteins / chemistry
  • Bacterial Outer Membrane Proteins / metabolism*
  • Bacterial Outer Membrane Proteins / ultrastructure
  • Cell Line
  • Cell Survival
  • Chaperonin 60 / chemistry
  • In Vitro Techniques
  • Microscopy, Electron, Transmission
  • Piscirickettsia / genetics
  • Piscirickettsia / metabolism*
  • Piscirickettsia / pathogenicity
  • Proteome / genetics
  • Virulence Factors / genetics
  • Virulence Factors / metabolism*

Substances

  • Bacterial Outer Membrane Proteins
  • Chaperonin 60
  • Proteome
  • Virulence Factors