Inflammasomes are multiprotein complexes that can be activated by a wide array of infectious and inflammatory agents. Inflammasome activation culminates in the maturation and secretion of pro-inflammatory cytokines, as well as lytic cell death, known as pyroptosis. During pyroptosis, the entire contents of a cell are released into the extracellular space, propagating the local innate immune response. One component of particular interest is the alarmin high mobility group box-1 (HMGB1). Extracellular HMGB1 is a potent inflammatory stimulus, acting upon multiple receptors to drive inflammation. In this series of protocols, we will outline how to trigger and assess pyroptosis in primary macrophages, with a focus on the assessment of HMGB1 release.
Keywords: ELISA; HMGB1; LDH assay; Macrophages; Pyroptosis; Western blot.
© 2023. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.