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Updated: Jul 17, 2025
Activation and Measurement of NLRP3 Inflammasome Activity Using IL-1β in Human Monocyte-derived Dendritic Cells
Published on: May 22, 2014
Protocol for in vivo and in vitro activation of NLRP3 inflammasome in mice using monosodium urate
Abstract:
Although intraperitoneal injection of monosodium urate (MSU) is an effective model for studying peritonitis, its establishment remains challenging. Here, we present a protocol for using MSU to activate the NLRP3 inflammasome in bone-marrow-derived macrophages (BMDMs) and to induce peritonitis in mice. We describe steps for isolating and culturing BMDMs, preparing MSU crystals, and activating the NLRP3 inflammasome using western blot and ELISA. We then detail procedures for inducing peritonitis and testing for relevant indicators using flow cytometry and ELISA. For complete details on the use and execution of this protocol, please refer to Huang et al. (2023).1.
Insights
This study provides a detailed protocol for activating the NLRP3 inflammasome in bone-marrow-derived macrophages (BMDMs) and inducing peritonitis in mice using monosodium urate (MSU) crystals.
Area of Science:
- Immunology
- Inflammation Research
- Molecular Biology
Background:
- The monosodium urate (MSU) crystal model is crucial for studying peritonitis.
- Establishing this model for peritonitis research presents significant challenges.
Purpose of the Study:
- To present a standardized protocol for activating the NLRP3 inflammasome in bone-marrow-derived macrophages (BMDMs).
- To detail a method for inducing peritonitis in mice using MSU crystals.
Main Methods:
- Isolation and culture of BMDMs.
- Preparation of MSU crystals and NLRP3 inflammasome activation.
- Induction of peritonitis and analysis using western blot, ELISA, and flow cytometry.
Main Results:
- Successful activation of the NLRP3 inflammasome in BMDMs.
- Effective induction of peritonitis in a mouse model.
- Quantification of inflammatory indicators via established assays.
Conclusions:
- The presented protocol offers a reliable method for studying MSU-induced peritonitis.
- This standardized approach facilitates research into NLRP3 inflammasome activation and inflammatory responses.
