{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"institution":[{"name":"bioRxiv"}],"indexed":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T10:18:22Z","timestamp":1785406702214,"version":"3.56.0"},"posted":{"date-parts":[[2025,5,30]]},"group-title":"Immunology","reference-count":33,"publisher":"openRxiv","license":[{"start":{"date-parts":[[2025,5,30]],"date-time":"2025-05-30T00:00:00Z","timestamp":1748563200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"}],"funder":[{"award":["R01AI55182"],"award-info":[{"award-number":["R01AI55182"]}],"id":[{"id":"https:\/\/ror.org\/01cwqze88","id-type":"ROR","asserted-by":"publisher"}]},{"name":"Medical Research Council","award":["MR\/W020556\/1"],"award-info":[{"award-number":["MR\/W020556\/1"]}]},{"award":["SGL021\/1030"],"award-info":[{"award-number":["SGL021\/1030"]}],"id":[{"id":"https:\/\/ror.org\/00c489v88","id-type":"ROR","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100000833","name":"Rosetrees Trust","doi-asserted-by":"crossref","award":["A2903"],"award-info":[{"award-number":["A2903"]}],"id":[{"id":"10.13039\/501100000833","id-type":"DOI","asserted-by":"crossref"}]},{"name":"Medical Research Foundation","award":["MRF-044-0004-F-GILL-C0823"],"award-info":[{"award-number":["MRF-044-0004-F-GILL-C0823"]}]},{"name":"Medical Research Council","award":["MR\/X020843\/1"],"award-info":[{"award-number":["MR\/X020843\/1"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"accepted":{"date-parts":[[2025,5,30]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                <jats:sec>\n                  <jats:title>Background<\/jats:title>\n                  <jats:p>Hepatitis B virus (HBV) infection remains a significant global health challenge, leading to chronic liver disease and hepatocellular carcinoma (HCC). Natural killer (NK) cells play an important role in the clearance of HBV-infected cells, but their efficacy is often compromised during chronic infection. Adaptive NK cells, characterised by NKG2C expression and enhanced functional responses, represent a promising therapeutic avenue for enhancing anti-HBV immunity and responses to HBV-driven cancers.<\/jats:p>\n                <\/jats:sec>\n                <jats:sec>\n                  <jats:title>Methods<\/jats:title>\n                  <jats:p>We applied an established protocol, involving K562-HLA-E expressing feeder cells and cytokines (IL-2), for the expansion of adaptive NK cells from cryopreserved T- and B cell depleted peripheral blood mononuclear cells (PBMCs) derived from donors with chronic HBV infection alone or with Human Immunodeficiency Virus (HIV) co-infection. We evaluated the adaptive profile of expanded NK cells, their antibody-dependent cellular cytotoxicity (ADCC) capacity and functional responses against hepatoma cell lines in the presence or absence of HBV infection.<\/jats:p>\n                <\/jats:sec>\n                <jats:sec>\n                  <jats:title>Results<\/jats:title>\n                  <jats:p>\n                    Expanded NK cells achieved &gt;97% purity, with the NKG2C positive population exhibiting a mean 100-fold expansion. These cells demonstrated a predominantly adaptive phenotype with high surface expression of NKG2C and cytotoxic potential (Granzyme B). They maintained high levels of CD16 surface expression and upregulated CD2, essential for ADCC. Functionally, expanded adaptive NK cells showed enhanced ADCC capacity and functional responses to K562 targets, naive, HBV integrant-expressing, and\n                    <jats:italic>de novo<\/jats:italic>\n                    infected hepatoma cell lines. TGF-\u03b2 preconditioning induced tissue-resident features (CD103, CD49a) in expanded adaptive NK cells, while preserving their adaptive phenotype and functionality, enhancing their potential for liver targeted immunotherapy. Further, expanded adaptive NK cells demonstrated minimal reactivity against autologous activated T cells, suggesting limited off-target effects.\n                  <\/jats:p>\n                <\/jats:sec>\n                <jats:sec>\n                  <jats:title>Conclusions<\/jats:title>\n                  <jats:p>Our study demonstrates the first successful expansion of adaptive NK cells with robust functional responses from donors with chronic viral infection. This approach creates opportunities for NK cell-based therapies alone or in combination with monoclonal antibodies contributing to HBV functional cure strategies and the treatment of HBV-driven cancers.<\/jats:p>\n                <\/jats:sec>","DOI":"10.1101\/2025.05.29.655595","type":"posted-content","created":{"date-parts":[[2025,5,30]],"date-time":"2025-05-30T21:10:19Z","timestamp":1748639419000},"source":"Crossref","is-referenced-by-count":2,"title":["Expanded adaptive NKG2C+ NK cells exhibit potent ADCC and functional responses against HBV-infected hepatoma cell lines"],"prefix":"10.64898","author":[{"ORCID":"https:\/\/orcid.org\/0009-0000-3773-492X","authenticated-orcid":false,"given":"Jonida","family":"Koki\u00e7i","sequence":"first","affiliation":[{"name":"Division of Infection and Immunity, UCL, London"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0001-3931-8636","authenticated-orcid":false,"given":"Helena","family":"Arellano-Ballestero","sequence":"additional","affiliation":[{"name":"Cancer Institute, UCL, London"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Benjamin","family":"Hammond","sequence":"additional","affiliation":[{"name":"INmune Bio International, Royal Free Hospital, London, UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7428-0058","authenticated-orcid":false,"given":"Anucha","family":"Preechanukul","sequence":"additional","affiliation":[{"name":"Division of Infection and Immunity, UCL, London"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Noshin","family":"Hussain","sequence":"additional","affiliation":[{"name":"Division of Infection and Immunity, UCL, London"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kelly","family":"da Costa","sequence":"additional","affiliation":[{"name":"Division of Infection and Immunity, UCL, London"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jessica","family":"Davies","sequence":"additional","affiliation":[{"name":"Division of Infection and Immunity, UCL, London"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sadiyah","family":"Mukhtar","sequence":"additional","affiliation":[{"name":"Mortimer Market Centre, Department of HIV, CNWL NHS Trust, London, United Kingdom"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Thomas","family":"Fernandez","sequence":"additional","affiliation":[{"name":"Royal Free London NHS Foundation Trust, London"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sabine","family":"Kinloch","sequence":"additional","affiliation":[{"name":"Royal Free London NHS Foundation Trust, London"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fiona M","family":"Burns","sequence":"additional","affiliation":[{"name":"Royal Free London NHS Foundation Trust, London"},{"name":"Institute for Global Health, UCL, London"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Patrick","family":"Kennedy","sequence":"additional","affiliation":[{"name":"Barts Liver Centre, Centre for Immunobiology, Barts and the London School of Medicine & Dentistry, QMUL, London"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Douglas","family":"MacDonald","sequence":"additional","affiliation":[{"name":"Royal Free London NHS Foundation Trust, London"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6384-1462","authenticated-orcid":false,"given":"Mala K","family":"Maini","sequence":"additional","affiliation":[{"name":"Division of Infection and Immunity, UCL, London"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Upkar S","family":"Gill","sequence":"additional","affiliation":[{"name":"Barts Liver Centre, Centre for Immunobiology, Barts and the London School of Medicine & Dentistry, QMUL, London"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alan Xiaodong","family":"Zhuang","sequence":"additional","affiliation":[{"name":"Division of Infection and Immunity, UCL, London"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2600-5024","authenticated-orcid":false,"given":"Mark W","family":"Lowdell","sequence":"additional","affiliation":[{"name":"Division of Infection and Immunity, UCL, London"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8718-9373","authenticated-orcid":false,"given":"Karl-Johan","family":"Malmberg","sequence":"additional","affiliation":[{"name":"Center for Infectious Medicine, Department of Medicine 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