Related Experiment Video
Updated: Jul 15, 2025

Detection of Inflammasome Activation and Pyroptotic Cell Death in Murine Bone Marrow-derived Macrophages
Published on: May 21, 2018
Sequentially activated death complexes regulate pyroptosis and IL-1β release in response to Yersinia blockade of
Ronit Schwartz Wertman1, Christina K Go1, Benedikt S Saller2,3
1Department of Pathobiology, University of Pennsylvania School of Veterinary Medicine, Philadelphia, Pennsylvania, USA 19104.
Abstract:
The Yersinia virulence factor YopJ potently inhibits immune signaling in macrophages by blocking activation of the signaling kinases TAK1 and IKK. In response, macrophages trigger a backup pathway of host defense that mediates cell death via the apoptotic enzyme caspase-8 and pyroptotic enzyme caspase-1. While caspase-1 is normally activated within multiprotein inflammasome complexes that contain the adaptor ASC and NLRs, which act as sensors of pathogen virulence, caspase-1 activation following Yersinia blockade of TAK1/IKK surprisingly requires caspase-8 and is independent of all known inflammasome components. Here, we report that caspase-1 activation by caspase-8 requires both caspase-8 catalytic and auto-processing activity. Intriguingly, while caspase-8 serves as an essential initiator of caspase-1 activation, caspase-1 amplifies its own activation through a feed-forward loop involving auto-processing, caspase-1-dependent cleavage of the pore-forming protein GSDMD, and subsequent activation of the canonical NLRP3 inflammasome. Notably, while caspase-1 activation and cell death are independent of inflammasomes during Yersinia infection, IL-1β release requires the canonical NLPR3 inflammasome. Critically, activation of caspase-8 and activation of the canonical inflammasome are kinetically and spatially separable events, as rapid capase-8 activation occurs within multiple foci throughout the cell, followed by delayed subsequent assembly of a single canonical inflammasome. Importantly, caspase-8 auto-processing normally serves to prevent RIPK3/MLKL-mediated necroptosis, and in caspase-8's absence, MLKL triggers NLPR3 inflammasome activation and IL-1β release. Altogether, our findings reveal that functionally interconnected but temporally and spatially distinct death complexes differentially mediate pyroptosis and IL-1β release to ensure robust host defense against pathogen blockade of TAK1 and IKK.
Insights
Yersinia
Area of Science:
- Immunology
- Cell Biology
- Microbial Pathogenesis
Background:
- Yersinia virulence factor YopJ inhibits macrophage immune signaling by blocking TAK1 and IKK kinases.
- Macrophages activate backup pathways involving caspase-8 and caspase-1 for host defense.
- Caspase-1 activation typically requires inflammasomes, but Yersinia infection bypasses this.
Approach:
- Investigated the mechanism of caspase-1 activation independent of inflammasomes during Yersinia infection.
- Examined the role of caspase-8 catalytic and auto-processing activity in caspase-1 activation.
- Analyzed the interplay between caspase-8, caspase-1, GSDMD, and the NLRP3 inflammasome.
Key Points:
- Caspase-1 activation by caspase-8 requires caspase-8's catalytic and auto-processing functions.
- Caspase-1 amplifies its own activation via a feed-forward loop, leading to pyroptosis.
- IL-1β release, unlike pyroptosis, ultimately requires the NLRP3 inflammasome, with distinct kinetics and spatial organization from initial caspase-8 activation.
Conclusions:
- Yersinia infection triggers caspase-8-dependent pyroptosis independently of inflammasomes.
- Caspase-1 amplifies pyroptosis and IL-1β release, with the latter dependent on NLRP3 inflammasome assembly.
- Distinct death complexes mediate pyroptosis and IL-1β release, ensuring host defense against Yersinia's immune evasion.
Related Concept Videos
The Extrinsic Apoptotic Pathway
Overview of Cell Death
Cell death was observed in the early 19th century, but there was no experimental evidence to prove it. In 1842, Carl Vogt first discovered cell death in a metamorphic toad; however, it was not termed ‘cell death.’ Scientists discovered different cell death pathways only in the...
The JAK-STAT Signaling Pathway
The Intrinsic Apoptotic Pathway
Caspases
Receptor Downregulation in MVBs
The EGFR can initiate signaling pathways that lead to cell proliferation, migration, and differentiation. Overexpression of EGFR stimulates cells to proliferate. Excessive EGFR...

