Topping, J., Lara-Reyna, S., Ibbotson, A. et al. (6 more authors) (2024) FACS-based detection of extracellular ASC specks from NLRP3 inflammasomes in inflammatory diseases. Clinical & Experimental Immunology, 219 (1). uxae117. ISSN 0009-9104
Abstract
Introduction The apoptosis-associated speck-like protein containing a caspase recruitment domain (ASC) is crucial for inflammasome assembly and activation of several inflammasomes, including NLRP3. ASC aggregates are detected in human sera post pyroptotic cell death, but their inflammasome origin remains unclear.
Method This study aimed to develop a method to detect ASC aggregates originating from NLRP3 inflammasomes. Initially, human monocytes, macrophages, and THP-1 ASC reporter cells were employed to validate the detection of ASC/NLRP3-positive events through flow cytometry.
Results The presence of ASC/NLRP3 specks was confirmed in cell supernatants from monocytes and macrophages treated with LPS and nigericin or ATP. Flow cytometry analysis identified double-positive specks in patient sera from inflammatory conditions when compared with healthy controls. Elevated ASC/NLRP3 specks were observed in conditions such as cryopyrin-associated periodic syndrome and Schnitzler’s syndrome.
Conclusion We validated fluorescence-activated cell sorting as a reliable method for detecting ASC/NLRP3 specks in human sera, with potential diagnostic and monitoring applications in certain systemic autoinflammatory diseases.
Metadata
Item Type: | Article |
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Authors/Creators: |
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Copyright, Publisher and Additional Information: | © The Author(s) 2024. This is an open access article under the terms of the Creative Commons Attribution-NonCommercial-NoDerivatives licence (CC-BY-NC-ND 4.0). |
Keywords: | inflammasome, NLRP3, ASC, inflammation, SAID |
Dates: |
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Institution: | The University of Leeds |
Academic Units: | The University of Leeds > Faculty of Medicine and Health (Leeds) > School of Medicine (Leeds) |
Funding Information: | Funder Grant number Leeds Hospitals Charity 5T02 & 4M04/A2001064 |
Depositing User: | Symplectic Publications |
Date Deposited: | 11 Mar 2025 10:48 |
Last Modified: | 11 Mar 2025 10:48 |
Status: | Published online |
Publisher: | Oxford University Press |
Identification Number: | 10.1093/cei/uxae117 |
Related URLs: | |
Open Archives Initiative ID (OAI ID): | oai:eprints.whiterose.ac.uk:224236 |