HBsAg-induced ROS drives NK cell exhaustion via LAG-3 upregulation and hinders HBeAg seroconversion in CHB

HBsAg-induced ROS drives NK cell exhaustion via LAG-3 upregulation and hinders HBeAg seroconversion in CHB | 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 HBsAg-induced ROS drives NK cell exhaustion via LAG-3 upregulation and hinders HBeAg seroconversion in CHB Junchi Xu, Ping Xu, Longji Cheng, Xing Lv, Hui Chen, Rui Qiang, and 2 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-8677306/v1 This work is licensed under a CC BY 4.0 License Status: Under Review Version 1 posted You are reading this latest preprint version Abstract Dysfunction of NK cells contributes to immune escape in chronic hepatitis B (CHB), yet the regulatory mechanisms, particularly the interplay between immune checkpoints and oxidative stress, remain incompletely understood. This study analyzed 42 CHB patients and 22 healthy controls, revealing a specific elevation in reactive oxygen species (ROS) levels within peripheral blood NK cells from CHB patients. These elevated ROS levels correlated positively with serum HBsAg, HBeAg, HBV-DNA loads, and ALT levels. Further transcriptomic and flow cytometric analyses demonstrated significant upregulation of the immune checkpoint molecule LAG-3 on NK cells from CHB patients, with the LAG-3⁺ NK subset exhibiting higher ROS levels. Mechanistically, in vitro experiments confirmed that HBsAg directly induced ROS burst and upregulated LAG-3 expression in NK cells, a process reversible by the antioxidant N-acetylcysteine. Clinical correlation analysis indicated that NK cell ROS levels predicted HBeAg seroconversion, with a significantly higher success rate in the low-ROS group. Our findings unveil a critical "HBsAg–ROS–LAG-3" axis in CHB immune evasion, providing a novel perspective for combined therapeutic strategies targeting both metabolism and immune checkpoints. Biological sciences/Immunology/Infectious diseases/Hepatitis/Viral hepatitis/Hepatitis B Biological sciences/Microbiology/Virology/Hepatitis B virus Full Text Additional Declarations There is NO Competing Interest. Cite Share Download PDF Status: Under Review Version 1 posted 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. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. 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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-8677306","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":584827923,"identity":"a9dc5156-21f5-4d82-9a0b-4359ac619427","order_by":0,"name":"Junchi 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