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Updated: Feb 6, 2026

Detection of Inflammasome Activation and Pyroptotic Cell Death in Murine Bone Marrow-derived Macrophages
06:52

Detection of Inflammasome Activation and Pyroptotic Cell Death in Murine Bone Marrow-derived Macrophages

Published on: May 21, 2018

11.3K

Caspase-11 auto-proteolysis is crucial for noncanonical inflammasome activation

Bettina L Lee1, Irma B Stowe1, Aaron Gupta1

  • 1Department of Physiological Chemistry, Genentech Inc., South San Francisco, CA.

The Journal of Experimental Medicine
|August 24, 2018
PubMed

Insights

Caspase-11 auto-cleavage is crucial for processing gasdermin D (GSDMD), a key step in pyroptosis and defense against Gram-negative bacteria. This self-cleavage regulates inflammatory responses and cell death pathways.

Area of Science:

  • Immunology
  • Cell Biology
  • Microbiology

Background:

  • Intracellular lipopolysaccharide (LPS) detection by caspase-4/5/11 initiates pyroptosis via gasdermin D (GSDMD) cleavage.
  • Caspase-4/5/11 and GSDMD are critical in sepsis and inflammatory diseases, making them important drug targets.

Purpose of the Study:

  • To investigate the role of caspase-11 auto-cleavage in its catalytic activity and GSDMD processing.
  • To confirm the indispensable role of the GSDMD Asp276 residue in pyroptotic activation.

Main Methods:

  • Utilized knock-in (KI) mouse models, including caspase-11-processing dead KI (Casp11) and enzymatically dead KI (Casp11) mice.
  • Generated Gsdmd KI macrophages to assess GSDMD cleavage and inflammasome response.
  • Analyzed macrophage responses to cytoplasmic LPS and Gram-negative bacterial infection.

Main Results:

  • Caspase-11 auto-cleavage at the inter-subunit linker is essential for its optimal activity and subsequent GSDMD cleavage.
  • Macrophages from Casp11 KI mice showed defective auto-processing and attenuated responses to LPS and bacterial infection.
  • Gsdmd KI macrophages failed to cleave GSDMD, confirming Asp276 as a critical site for its activation and highlighting hypo-responsiveness to inflammasome stimuli.

Conclusions:

  • Caspase-11 self-cleavage is a critical regulatory step for GSDMD processing and the cellular defense response against Gram-negative bacteria.
  • The GSDMD Asp276 residue is indispensable for its proteolytic activation.
  • Findings provide genetic evidence for the regulatory mechanisms of pyroptosis and potential therapeutic targets.

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