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

• Published in: Methods in molecular biology (Clifton, N.J.) Vol. 2459; p. 51
• Main Authors: Bezbradica, Jelena S, Coll, Rebecca C, Boucher, Dave
• Format: Journal Article
• Language: English
• Published: United States 2022
• Subjects: Animals; Humans; Inflammasomes - metabolism; Intracellular Signaling Peptides and Proteins - metabolism; Mice; Neoplasm Proteins - metabolism; Phosphate-Binding Proteins; Pyroptosis; Monocytes; Pyroptosis; Caspase-11; Epithelial cells; Caspase; Lactate dehydrogenase; Gasdermin D; Caspase-4; Macrophages; Non-canonical inflammasome; Lipopolysaccharides
• ISSN: 1940-6029
• DOI: 10.1007/978-1-0716-2144-8_5

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.