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Updated: Mar 8, 2026

Activation and Measurement of NLRP3 Inflammasome Activity Using IL-1β in Human Monocyte-derived Dendritic Cells
Published on: May 22, 2014
MLKL Activation Triggers NLRP3-Mediated Processing and Release of IL-1β Independently of Gasdermin-D
Kimberley D Gutierrez1, Michael A Davis1, Brian P Daniels1
1Department of Immunology, University of Washington, Seattle, WA 98109.
Abstract:
Necroptosis is a form of programmed cell death defined by activation of the kinase receptor interacting protein kinase 3 and its downstream effector, the pseudokinase mixed lineage kinase domain-like (MLKL). Activated MLKL translocates to the cell membrane and disrupts it, leading to loss of cellular ion homeostasis. In this study, we use a system in which this event can be specifically triggered by a small-molecule ligand to show that MLKL activation is sufficient to induce the processing and release of bioactive IL-1β. MLKL activation triggers potassium efflux and assembly of the NLRP3 inflammasome, which is required for the processing and activity of IL-1β released during necroptosis. Notably, MLKL activation also causes cell membrane disruption, which allows efficient release of IL-1β independently of the recently described pyroptotic effector gasdermin-D. Taken together, our findings indicate that MLKL is an endogenous activator of the NLRP3 inflammasome, and that MLKL activation provides a mechanism for concurrent processing and release of IL-1β independently of gasdermin-D.
Insights
Mixed lineage kinase domain-like (MLKL) activation during necroptosis triggers IL-1β release. This process involves potassium efflux and NLRP3 inflammasome assembly, independent of gasdermin-D.
Area of Science:
- Cellular Biology
- Immunology
- Molecular Biology
Background:
- Necroptosis is programmed cell death initiated by RIPK3 and MLKL.
- Activated MLKL disrupts the cell membrane, causing ion imbalance.
- Interleukin-1 beta (IL-1β) is a key inflammatory cytokine.
Purpose of the Study:
- To investigate if MLKL activation alone can induce IL-1β processing and release.
- To elucidate the role of MLKL in IL-1β release during necroptosis.
- To determine the mechanism of IL-1β release independent of gasdermin-D.
Main Methods:
- Utilized a small-molecule ligand to specifically trigger MLKL activation.
- Measured IL-1β processing and release.
- Assessed potassium efflux and NLRP3 inflammasome assembly.
- Investigated MLKL's role in membrane disruption and IL-1β release.
Main Results:
- MLKL activation was sufficient to process and release bioactive IL-1β.
- MLKL activation induced potassium efflux and NLRP3 inflammasome assembly.
- Cell membrane disruption by MLKL facilitated IL-1β release independently of gasdermin-D.
- MLKL acts as an endogenous activator of the NLRP3 inflammasome.
Conclusions:
- MLKL activation is a key driver for IL-1β processing and release during necroptosis.
- MLKL-mediated membrane disruption provides a pathway for IL-1β release independent of gasdermin-D.
- This study identifies MLKL as a crucial component in regulating IL-1β secretion via the NLRP3 inflammasome.
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