Related Experiment Video
Updated: Dec 6, 2025
06:52
Detection of Inflammasome Activation and Pyroptotic Cell Death in Murine Bone Marrow-derived Macrophages
Published on: May 21, 2018
11.1K
Inflammasome Assays In Vitro and in Mouse Models
Haitao Guo1, Jenny P-Y Ting1,2,3
1Department of Genetics, University of North Carolina at Chapel Hill, Chapel Hill, North Carolina.
Current Protocols in Immunology
|October 5, 2020
Summary
This study details assays for inflammasome activation and measurement in immune cells and animal models. These methods enable the study of key inflammatory responses, including interleukin-1β and IL-18 maturation and pyroptosis.
Area of Science:
- Immunology
- Molecular Biology
- Cell Biology
Background:
- Inflammasomes are critical protein complexes in innate immunity, regulating inflammatory responses.
- Dysregulated inflammasome activation is implicated in various inflammatory diseases.
- Standardized assays are needed to study inflammasome activation and its consequences.
Purpose of the Study:
- To present a comprehensive set of assays for inducing and measuring inflammasome activation.
- To provide protocols for studying different inflammasomes (NLRP1, NLRP3, NLRC4, AIM2) in various experimental systems.
- To enable the assessment of inflammasome-mediated outcomes like cytokine maturation and pyroptosis.
Main Methods:
- Priming and stimulation of inflammasomes in mouse macrophages and human PBMCs.
- Induction of inflammasome activation in mouse models of peritonitis and endotoxic shock.
- Purification, detection, and immunofluorescence staining of ASC pyroptosomes.
- Measurement of IL-1β, IL-18, and other cytokines in cellular and serum samples.
Main Results:
- Established protocols for activating specific inflammasomes using defined stimuli.
- Demonstrated measurement of inflammasome-dependent IL-1β and IL-18 maturation and pyroptosis.
- Validated methods for detecting ASC pyroptosome formation as a marker of inflammasome activation.
- Successfully induced and analyzed inflammasome-driven peritonitis and sepsis models in mice.
Conclusions:
- The presented assays provide a robust framework for studying inflammasome biology.
- These methods facilitate the investigation of inflammasome roles in innate immunity and disease.
- The protocols are applicable across different cell types and in vivo models, offering versatility for researchers.
