Inflammasome activation and inhibition in primary murine bone marrow-derived cells, and assays for IL-1α, IL-1β, and caspase-1

Methods Mol Biol. 2013:1040:117-35. doi: 10.1007/978-1-62703-523-1_10.

Abstract

Through its ability to control the proteolytic maturation and secretion of interleukin-1 family cytokines, the inflammasome occupies a central role in the activation of inflammation and also influences the shaping of adaptive immunity. Since it affects a multitude of different immune responses from autoinflammatory diseases to host defense, vaccine efficacy, and even cancer, it has become of interest to many researchers. Here, we describe a straightforward method for inflammasome assays in primary murine bone marrow--derived myeloid cells. The protocol encompasses cell handling, inflammasome activation and inhibition, as well as the detection of IL-1β, caspase-1, and IL-1α by ELISA and Western blot.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Blotting, Western
  • Bone Marrow Cells / immunology
  • Bone Marrow Cells / metabolism*
  • Caspase 1 / metabolism*
  • Dendritic Cells / immunology
  • Dendritic Cells / metabolism
  • Enzyme-Linked Immunosorbent Assay
  • Inflammasomes / immunology
  • Inflammasomes / metabolism*
  • Interleukin-1 / metabolism*
  • Interleukin-1alpha / metabolism
  • Interleukin-1beta / metabolism
  • Mice

Substances

  • Inflammasomes
  • Interleukin-1
  • Interleukin-1alpha
  • Interleukin-1beta
  • Caspase 1