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

  • Helios Martínez-Banaclocha
  • Pablo Pelegrín
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 2041)

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.

Key words

NLRP3 Flow cytometry ASC Monocyte Inflammasome ATP P2X7 

Notes

Acknowledgments

This work was supported by the European Research Council (ERC-2013-CoG 614578), EU COST Program (BM1406), Fundación Séneca (20859/PI/18) and Ministerio de Economia, Industria y Competitividad–Fondo Europeo de Desarrollo Regional (project no. SAF2017-88276-R).

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2020

Authors and Affiliations

  • Helios Martínez-Banaclocha
    • 1
  • Pablo Pelegrín
    • 1
  1. 1.Biomedical Research Institute of Murcia (IMIB)Clinical University Hospital Virgen de la ArrixacaMurciaSpain

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