Detection of Inflammasome Activation by P2X7 Purinoceptor Activation by Determining ASC Oligomerization

Methods Mol Biol. 2020:2041:335-343. doi: 10.1007/978-1-4939-9717-6_25.

Abstract

The purinoceptor P2X7 is highly expressed in cells of the innate immune system, including monocytes and macrophages. Its activation is a potent signal to activate the NLRP3 inflammasome and induce the release of proinflammatory cytokines of the IL-1 family. In this chapter, we present a method to monitor NLRP3 inflammasome activation in human monocytes upon P2X7 receptor stimulation by detecting intracellular oligomers of ASC by flow cytometry. This method could be used to evaluate the degree of inflammasome activation in blood samples from patients suffering different chronic inflammatory diseases.

Keywords: ASC; ATP; Flow cytometry; Inflammasome; Monocyte; NLRP3; P2X7.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenosine Triphosphate / metabolism*
  • CARD Signaling Adaptor Proteins / chemistry*
  • Cells, Cultured
  • Cytokines / metabolism
  • Flow Cytometry
  • Humans
  • Inflammasomes / metabolism*
  • Lipopolysaccharides
  • Monocytes / cytology
  • Monocytes / metabolism*
  • NLR Family, Pyrin Domain-Containing 3 Protein / metabolism*
  • Protein Multimerization*
  • Receptors, Purinergic P2X7 / metabolism*
  • Signal Transduction

Substances

  • CARD Signaling Adaptor Proteins
  • Cytokines
  • Inflammasomes
  • Lipopolysaccharides
  • NLR Family, Pyrin Domain-Containing 3 Protein
  • NLRP3 protein, human
  • PYCARD protein, human
  • Receptors, Purinergic P2X7
  • Adenosine Triphosphate