Outer membrane vesicles prime and activate macrophage inflammasomes and cytokine secretion in vitro and in vivo

Jessica D. Cecil, Neil M O'Brien-Simpson, Jason C Lenzo, James A Holden, William C J Singleton, Alexis Perez-Gonzalez, Ashley Mansell, Eric Charles Reynolds

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28 Citations (Scopus)

Abstract

Outer membrane vesicles (OMVs) are proteoliposomes blebbed from the surface of Gram-negative bacteria. Chronic periodontitis is associated with an increase in subgingival plaque of Gram-negative bacteria, Porphyromonas gingivalis, Treponema denticola, and Tannerella forsythia. In this study, we investigated the immune-modulatory effects of P. gingivalis, T. denticola, and T. forsythia OMVs on monocytes and differentiated macrophages. All of the bacterial OMVs were phagocytosed by monocytes, M(naïve) and M(IFNγ) macrophages in a dose-dependent manner. They also induced NF-κB activation and increased TNFα, IL-8, and IL-1β cytokine secretion. P. gingivalis OMVs were also found to induce anti-inflammatory IL-10 secretion. Although unprimed monocytes and macrophages were resistant to OMV-induced cell death, lipopolysaccharide or OMV priming resulted in a significantly reduced cell viability. P. gingivalis, T. denticola, and T. forsythia OMVs all activated inflammasome complexes, as monitored by IL-1beta; secretion and ASC speck formation. ASC was critical for OMV-induced inflammasome formation, while AIM2-/- and Caspase-1-/- cells had significantly reduced inflammasome formation and NLRP3-/- cells exhibited a slight reduction. OMVs were also found to provide both priming and activation of the inflammasome complex. High-resolution microscopy and flow cytometry showed that P. gingivalis OMVs primed and activated macrophage inflammasomes in vivo with 80% of macrophages exhibiting inflammasome complex formation. In conclusion, periodontal pathogen OMVs were found to have significant immunomodulatory effects upon monocytes and macrophages and should therefore influence pro-inflammatory host responses associated with disease.

Original languageEnglish
Article number1017
Number of pages22
JournalFrontiers in Immunology
Volume8
Issue numberAUG
DOIs
Publication statusPublished - 25 Aug 2017

Keywords

  • Inflammasomes
  • Macrophages
  • Outer membrane vesicles
  • Periodontitis
  • Porphyromonas gingivalis
  • Tannerella forsythia
  • Treponema denticola

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