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Detection of Gasdermin Activation and Lytic Cell Death During Pyroptosis and Apoptosis

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Effector-Triggered Immunity

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2523))

Abstract

Cytosolic pattern recognition receptors trigger pyroptosis by detection of danger- or pathogen-associated molecular patterns. These receptors initiate the assembly of inflammasomes, multimeric protein complexes that drive caspase-1 activation. Active caspase-1 cleaves the proinflammatory cytokines IL-1β and IL-18 and the pore-forming protein gasdermin-D (GSDMD) thereby liberating its N-terminal domain. The GSDMD N-termini form multimeric pores at the plasma membrane that allow leakage of intracellular content and ultimately trigger a type of cell death called “pyroptosis.” Emerging studies have revealed that GSDMD is also processed by apoptotic caspases-8/-3/-7. In this chapter, we aim to describe methods to monitor lytic cell death and to distinguish between GSDMD processing events and the GSDMD fragments that are generated after pyroptosis or apoptosis induction. We also illustrate the difference between GSDMD pore formation, and final cell lysis, and how this affects to the release of intracellular content. Finally, we show that the activation of another pore-forming protein, gasdermin-E, does not exclusively translate into lytic cell death in bone marrow-derived macrophages.

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Acknowledgments

This work was supported by the European Research Council grant (ERC2017-CoG-770988-InflamCellDeath) and a Swiss National Science Foundation grant (SNSF 175576).

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Authors and Affiliations

  1. Department of Biochemistry, University of Lausanne, Epalinges, Switzerland

    Benjamin Demarco, Saray Ramos & Petr Broz

  2. Kennedy Institute of Rheumatology, NDORMS, University of Oxford, Oxford, UK

    Benjamin Demarco

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  1. Benjamin Demarco
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  2. Saray Ramos
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  3. Petr Broz
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Correspondence to Petr Broz .

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  1. Department of Immunology, Institute of Nutritional Medicine, University of Hohenheim, Stuttgart, Baden-Württemberg, Germany

    Thomas A. Kufer

  2. Department of Physiology Anatomy & Microbiology, La Trobe University, Bundoora, VIC, Australia

    Maria Kaparakis-Liaskos

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Demarco, B., Ramos, S., Broz, P. (2022). Detection of Gasdermin Activation and Lytic Cell Death During Pyroptosis and Apoptosis. In: Kufer, T.A., Kaparakis-Liaskos, M. (eds) Effector-Triggered Immunity. Methods in Molecular Biology, vol 2523. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2449-4_14

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