Activation of the Non-canonical Inflammasome in Mouse and Human Cells

Jelena S Bezbradica, Rebecca C Coll, Dave Boucher

Research output: Chapter in Book/Report/Conference proceedingChapter (peer-reviewed)peer-review

Abstract

The non-canonical inflammasome is a signaling platform that allows for the detection of cytoplasmic lipopolysaccharides (LPS) in immune and non-immune cells. Upon detection of LPS, this inflammasome activates the signaling proteases caspase-4 and -5 (in humans) and caspase-11 (in mice). Inflammatory caspases activation leads to caspase self-processing and the cleavage of the pore-forming protein Gasdermin D (GSDMD). GSDMD N-terminal fragments oligomerize and form pores at the plasma membranes, leading to an inflammatory form of cell death called pyroptosis. Here, we describe a simple method to activate the non-canonical inflammasome in myeloid and epithelial cells and to measure its activity using cell death assay and immunoblotting.
Original languageEnglish
Title of host publicationThe Inflammasome: Methods and Protocols
EditorsAli A. Abdul-Sater
PublisherSpringer
Pages51-63
ISBN (Electronic)978-1-0716-2144-8
ISBN (Print)978-1-0716-2143-1
DOIs
Publication statusPublished - 01 Jan 2022

Publication series

NameMethods in molecular biology (Clifton, N.J.)
PublisherHumana Press
Volume 2459
ISSN (Print)1064-3745

Keywords

  • Humans
  • Caspase
  • Caspase-4
  • Gasdermin D
  • Epithelial cells
  • Macrophages
  • Phosphate-Binding Proteins
  • Lactate dehydrogenase
  • Pyroptosis
  • Intracellular Signaling Peptides and Proteins - metabolism
  • Lipopolysaccharides
  • Inflammasomes - metabolism
  • Monocytes
  • Mice
  • Neoplasm Proteins - metabolism
  • Caspase-11
  • Non-canonical inflammasome
  • Animals

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