Assays for Measuring the Role of MIF in NLRP3 Inflammasome Activation

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Abstract

Hallmarks of NLRP3 inflammasome activation include the cleavage and secretion of the mature forms of caspase-1, IL-1β, and IL-18 and aggregation of ASC into "specks." We have previously shown that macrophage migratory inhibitory factor (MIF) directly regulates activation of the NLRP3 inflammasome, inhibiting the release of interleukin (IL)-1α, IL-1β, and IL-18. Here we present protocols for studying activation of the NLRP3 inflammasome in human and mouse macrophages and peripheral blood mononuclear cells (PBMCs). These protocols can also be applied to different cell types, such as fibroblasts, neutrophils, endothelial cells, and epithelial cells, although further optimization may be required for each. We also cover the stimulation of macrophages with established NLRP3 inflammasome activators.

Original languageEnglish
Title of host publicationMacrophage Migration Inhibitory Factor
Subtitle of host publicationMethods and Protocols
EditorsJames Harris, Eric F. Morand
Place of PublicationNew York, NY
PublisherHumana Press
Chapter14
Pages159-172
Number of pages14
Volume2080
ISBN (Electronic)9781493999361
ISBN (Print)9781493999354
DOIs
Publication statusPublished - 2020

Publication series

NameMethods in Molecular Biology
PublisherHumana Press
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Keywords

  • ASC oligomerization
  • Caspase-1 activity
  • ELISA
  • Immunoblotting
  • Interleukin-18 activation
  • Interleukin-1α activation
  • Interleukin-1β activation
  • MIF
  • NLRP3
  • NLRP3 reporter assays

Cite this

Pinar, A. A., & Harris, J. (2020). Assays for Measuring the Role of MIF in NLRP3 Inflammasome Activation. In J. Harris, & E. F. Morand (Eds.), Macrophage Migration Inhibitory Factor: Methods and Protocols (Vol. 2080, pp. 159-172). (Methods in Molecular Biology). Humana Press. https://doi.org/10.1007/978-1-4939-9936-1_14