Sublethal Disulfide Stress Facilitates TLR2-Dependent NLRP3 Inflammasome Activation and Pyroptosis in THP-1 Cells

Sublethal Disulfide Stress Facilitates TLR2-Dependent NLRP3 Inflammasome Activation and Pyroptosis in THP-1 Cells | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Article Sublethal Disulfide Stress Facilitates TLR2-Dependent NLRP3 Inflammasome Activation and Pyroptosis in THP-1 Cells Kohsuke Tsuchiya, Shoko Hosojima, Shenghui Zhi, Yamato Tanabe, and 2 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-8485758/v1 This work is licensed under a CC BY 4.0 License Status: Under Review Version 1 posted 9 You are reading this latest preprint version Abstract Inflammasomes are innate immune signaling platforms that activate caspase-1, which in turn cleaves gasdermin D (GSDMD) into its mature form, leading to pyroptosis, an inflammatory form of regulated necrosis. It has been reported that bacterial lipopeptides (BLPs) trigger pyroptosis through NLRP7 inflammasome activation. However, the underlying recognition mechanism remains entirely unclear, and the concept of BLP sensing by NLRP7 has not been well established. In this study, we sought to elucidate the mechanism by which BLPs induce inflammasome activation and pyroptosis in THP-1 cells, a human acute myeloid leukemia cell line. Through comprehensive analyses, we demonstrate that BLPs induce GSDMD-dependent pyroptosis not via the NLRP7 inflammasome, but through activation of the NLRP3 inflammasome without the need for additional stimuli. Addition of KCl to the culture medium significantly suppressed BLP-induced pyroptosis, suggesting that K⁺ efflux is required for inflammasome activation by BLPs. However, BLPs alone are generally thought to be insufficient to directly induce K+ efflux, prompting us to hypothesize that K+ efflux might result from sublethal membrane damage associated with a distinct cell death mechanism. Among several cell death inhibitors tested, DTT, an inhibitor of disulfidptosis, which is necrotic cell death caused by excessive disulfide bonding in the actin cytoskeleton, markedly attenuated BLP-induced pyroptosis. Consistent with this, BLP stimulation led to NADPH depletion and increased disulfide bond formation in β-actin, hallmarks of disulfidptosis. These findings suggest that disulfide stress contributes to NLRP3 inflammasome activation and pyroptosis triggered by BLPs in THP-1 cells. To our knowledge, this is the first report linking disulfidptosis-associated disulfide stress to innate immune signaling, providing a conceptual framework for exploiting pyroptosis induction in leukemia cells. Biological sciences/Immunology/Cell death and immune response Biological sciences/Immunology/Innate immunity/Pattern recognition receptors/NOD-like receptors inflammasome Pyroptosis NLRP7 NLRP3 TLR2 Disulfidptosis Full Text Additional Declarations (Not answered) Supplementary Files OriginalRawDataWesternblots.docx Original Raw Data_Western blots SupplementaryInformation.docx Supplementary Information Cite Share Download PDF Status: Under Review Version 1 posted Editorial decision: revise 04 Feb, 2026 Review # 1 received at journal 02 Feb, 2026 Review # 2 received at journal 21 Jan, 2026 Reviewer # 2 agreed at journal 21 Jan, 2026 Reviewer # 1 agreed at journal 21 Jan, 2026 Reviewers invited by journal 21 Jan, 2026 Submission checks completed at journal 06 Jan, 2026 Editor assigned by journal 30 Dec, 2025 First submitted to journal 30 Dec, 2025 You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-8485758","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":570191308,"identity":"41b35f94-3a5e-48cd-9cfb-0a7a36b91019","order_by":0,"name":"Kohsuke 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