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Quantifying Cell Death Induced by the NLRC4 Inflammasome

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NLR Proteins

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2696))

Abstract

The Nod-like Receptor (NLR) apoptosis inhibitory proteins (NAIPs) are cytosolic receptors that sense cytosolic bacterial proteins. NAIP ligation induces its association with NLRC4, leading to the assembly of the NAIP/NLRC4 inflammasome, which induces the activation of the caspase-1 protease. Caspase-1 then cleaves pro-interleukin (IL)-1β, pro-IL-18, and gasdermin D and induces a form of pro-inflammatory cell death, pyroptosis. These processes culminate in host defense against bacterial infection. Here we describe methods for activating NAIP/NLRC4 inflammasome signalling in human and murine macrophages and quantifying inflammasome-induced cell death.

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Acknowledgments and Disclosures

This work was supported by the Australian Research Council (Discovery Project DP190102285 and DP200100646 to KS; Discovery Early Career Researcher Fellowships DE220100823 and DE200101300 to SE and MMM) and the National Health and Medical Research Council of Australia (Project grant 1163924 and Fellowship 1141131 to KS).

KS is a co-inventor on patent applications for NLRP3 inhibitors which have been licensed to Inflazome Ltd., a company headquartered in Dublin, Ireland. Inflazome is developing drugs that target the NLRP3 inflammasome to address unmet clinical needs in inflammatory disease. KS served on the Scientific Advisory Board of Inflazome in 2016–2017 and serves as a consultant to Quench Bio, USA, and Novartis, Switzerland. At the time this protocol was developed, HK, AS, JA, MAN, WL, and IK were employees of Quench Bio, a company focused on therapeutically targeting GSDMD.

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Authors and Affiliations

  1. Institute for Molecular Bioscience, and Centre for Inflammation and Disease Research, The University of Queensland, St. Lucia, Australia

    Stefan Emming, Mercedes M. Monteleone & Kate Schroder

  2. Quench Bio, Inc., Cambridge, MA, USA

    Hiroto Kambara, Alina Starchenko, Jennifer Alley, Michael A. Nolan, Wei Li & Iain Kilty

Authors
  1. Stefan Emming
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  2. Mercedes M. Monteleone
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  3. Hiroto Kambara
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  4. Alina Starchenko
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  5. Jennifer Alley
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  6. Michael A. Nolan
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  7. Wei Li
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  8. Iain Kilty
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  9. Kate Schroder
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Corresponding author

Correspondence to Kate Schroder .

Editor information

Editors and Affiliations

  1. Molecular Inflammation Group, Biomedical Research Institute of Murcia, El Palmar, Spain

    Pablo Pelegrín

  2. Department of Morphology, University of Ferrara, Ferrara, Ferrara, Italy

    Francesco Di Virgilio

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© 2023 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature

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Emming, S. et al. (2023). Quantifying Cell Death Induced by the NLRC4 Inflammasome. In: Pelegrín, P., Di Virgilio, F. (eds) NLR Proteins. Methods in Molecular Biology, vol 2696. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3350-2_13

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  • DOI: https://doi.org/10.1007/978-1-0716-3350-2_13

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-3349-6

  • Online ISBN: 978-1-0716-3350-2

  • eBook Packages: Springer Protocols

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