Porphyromonas gingivalis-impaired innate immune response via NLRP3 proteolysis in endothelial cells

Innate Immun. 2015 Jan;21(1):65-72. doi: 10.1177/1753425914523459. Epub 2014 Feb 28.

Abstract

Porphyromonas gingivalis (Pg) is involved in the link between periodontal diseases and atherosclerosis worsening. In periodontal cells, Pg modifies IL-1β expression via the NLRP3 inflammasome pathway activation. Our aim was to investigate NLRP3 inflammasome activation in endothelial cells (ECs) after Pg infection and Pg-LPS stimulation. In both situations, RT-PCR experiments demonstrated an increase of the NLRP3 mRNA level that can be potentiated by pre-treatment of ECs with 5 mM ATP. However, Western blotting analysis revealed that Pg infection induced a proteolysis of NLRP3 protein and a major decrease of the native protein. After ATP pre-treatment and/or Pg-LPS stimulation, this proteolysis was not observed, while NLRP3 protein levels were increased. Proteolysis of the NLRP3 protein was not observed with heat-killed Pg and inhibition of ECs protein synthesis with cycloheximide did not abolish the NLRP3 protein degradation induced by Pg infection in ATP pre-treated cells. Additionally, significant increases of secreted IL-1β were measured after ATP pre-treatment and/or Pg-LPS stimulation, but not after Pg infection. These data showed that Pg and Pg-LPS differentially controlled the NLRP3 inflammasome pathway in ECs, and suggested a novel potential mechanism developed by Pg to reduce IL-1β secretion and to escape host immune response.

Keywords: Inflammasome; NLRP3; Porphyromonas gingivalis; atherosclerosis; endothelial; inflammation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bacteroidaceae Infections / immunology*
  • Bacteroidaceae Infections / microbiology
  • Carrier Proteins / immunology*
  • Cells, Cultured
  • Cycloheximide / pharmacology
  • Endothelial Cells / immunology*
  • Human Umbilical Vein Endothelial Cells
  • Humans
  • Immunity, Innate / immunology*
  • Inflammasomes
  • Interleukin-1beta / metabolism
  • Lipopolysaccharides / immunology
  • NLR Family, Pyrin Domain-Containing 3 Protein
  • Porphyromonas gingivalis / immunology*
  • Protein Synthesis Inhibitors / pharmacology
  • RNA, Messenger / biosynthesis
  • RNA, Messenger / genetics
  • Signal Transduction / immunology

Substances

  • Carrier Proteins
  • Inflammasomes
  • Interleukin-1beta
  • Lipopolysaccharides
  • NLR Family, Pyrin Domain-Containing 3 Protein
  • NLRP3 protein, human
  • Protein Synthesis Inhibitors
  • RNA, Messenger
  • Cycloheximide