Related Experiment Video

Updated: Sep 25, 2025

Activation and Measurement of NLRP3 Inflammasome Activity Using IL-1β in Human Monocyte-derived Dendritic Cells
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Activation and Measurement of NLRP3 Inflammasome Activity Using IL-1β in Human Monocyte-derived Dendritic Cells

Published on: May 22, 2014

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Mitochondrial electron transport chain is necessary for NLRP3 inflammasome activation

Leah K Billingham1, Joshua S Stoolman1, Karthik Vasan1

  • 1Department of Medicine, Northwestern University Feinberg School of Medicine, Chicago, IL, USA.

Nature Immunology
|April 28, 2022
PubMed

Insights

Mitochondria regulate NLRP3 inflammasome activation through ATP generation, not reactive oxygen species. Inhibiting the electron transport chain depletes phosphocreatine, lowering ATP and preventing inflammation.

Area of Science:

  • Immunology
  • Cell Biology
  • Mitochondrial Biology

Background:

  • The NLRP3 inflammasome mediates inflammation in chronic diseases.
  • Mitochondria, particularly through reactive oxygen species (ROS), are implicated in NLRP3 inflammasome activation.

Purpose of the Study:

  • To investigate the role of mitochondrial electron transport chain (ETC) in NLRP3 inflammasome activation.
  • To determine if ROS are essential for mitochondrial regulation of NLRP3 inflammasome.

Main Methods:

  • Inhibition of mitochondrial ETC complexes (I, II, III, V) and assessment of NLRP3 activation.
  • Complementation of ETC complex function using Saccharomyces cerevisiae NADH dehydrogenase (NDI1) and Ciona intestinalis alternative oxidase.
  • Metabolomic analysis to identify key metabolites affected by ETC inhibition.
  • Measurement of ATP levels and their correlation with NLRP3 inflammasome activation.

Main Results:

  • Mitochondrial ETC inhibitors blocked NLRP3 inflammasome activation.
  • NDI1 and alternative oxidase expression rescued inflammasome activation without ROS generation.
  • Metabolomics identified phosphocreatine (PCr) depletion by ETC inhibitors.
  • PCr depletion led to decreased ATP levels and reduced NLRP3 inflammasome activation.

Conclusions:

  • The mitochondrial ETC sustains NLRP3 inflammasome activation via a ROS-independent mechanism.
  • Phosphocreatine (PCr) is crucial for maintaining ATP levels required for NLRP3 inflammasome activation.
  • Mitochondrial ATP production, supported by PCr, is a key regulator of NLRP3 inflammasome signaling.

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