{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,25]],"date-time":"2026-08-25T05:19:02Z","timestamp":1787635142588,"version":"build-2736575974"},"reference-count":34,"publisher":"MDPI AG","issue":"17","license":[{"start":{"date-parts":[[2026,8,23]],"date-time":"2026-08-23T00:00:00Z","timestamp":1787443200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100004587","name":"Instituto de Salud Carlos III","doi-asserted-by":"publisher","award":["PI21\/01628"],"award-info":[{"award-number":["PI21\/01628"]}],"id":[{"id":"10.13039\/501100004587","id-type":"DOI","asserted-by":"publisher"}]},{"award":["PI21\/01628"],"award-info":[{"award-number":["PI21\/01628"]}],"id":[{"id":"https:\/\/ror.org\/00ca2c886","id-type":"ROR","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100000780","name":"European Union","doi-asserted-by":"crossref","id":[{"id":"10.13039\/501100000780","id-type":"DOI","asserted-by":"crossref"}]},{"id":[{"id":"https:\/\/ror.org\/019w4f821","id-type":"ROR","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100002704","name":"Asociaci\u00f3n Espa\u00f1ola Contra el C\u00e1ncer","doi-asserted-by":"publisher","award":["PRDVA222361MESE"],"award-info":[{"award-number":["PRDVA222361MESE"]}],"id":[{"id":"10.13039\/501100002704","id-type":"DOI","asserted-by":"publisher"}]},{"award":["PRDVA222361MESE"],"award-info":[{"award-number":["PRDVA222361MESE"]}],"id":[{"id":"https:\/\/ror.org\/00gxct719","id-type":"ROR","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Cancers"],"abstract":"<jats:p>Background: Relapse and opportunistic viral infections remain major causes of treatment failure after allogeneic hematopoietic stem cell transplantation (allo-HSCT). Adoptive transfer of natural killer (NK) cells is a promising strategy to enhance post-transplant immune reconstitution. Adaptive NKG2C+ NK cells exhibit enhanced cytotoxicity and persistence, but the influence of baseline donor immunophenotype on GMP manufacturing has not been systematically investigated. We evaluated whether donor NKG2A\/NKG2C immunophenotypes are associated with successful manufacture of adaptive NK-cell products. Methods: Eighty-three healthy donors from the ReDoCel registry underwent immunophenotypic characterization of circulating NK-cell subsets by multiparametric flow cytometry. Donors were stratified by unsupervised hierarchical clustering according to NKG2A\/NKG2C expression. Representative donors from NKG2C- and NKG2A-dominant clusters underwent feeder-free GMP-compliant expansion using the automated CliniMACS Prodigy\u00ae platform (Miltenyi Biotec, Bergisch Gladbach, Germany). Expanded products were evaluated for manufacturing efficiency, immunophenotype, cytotoxic function, and post-thaw stability. Results: Baseline NKG2C frequencies showed marked inter-donor variability, allowing identification of four immunophenotypic clusters. Only the NKG2C-dominant donor achieved successful GMP manufacturing, exceeding the predefined expansion threshold while maintaining high viability and purity. Both NKG2A-dominant donors showed limited proliferative capacity under identical manufacturing conditions. Expanded NK cells acquired an activated phenotype characterized by increased expression of DNAM-1, NKG2D, NKp30, NKp46, and TIM-3 while preserving mature differentiation and KIR expression. Functional analyses demonstrated potent degranulation against leukemia targets with minimal autoreactivity, resulting in a predominantly cytotoxic effector profile. The successfully expanded product maintained viability, phenotype, and function after long-term cryopreservation. Conclusions: This proof-of-concept study suggests that baseline donor NKG2A\/NKG2C immunophenotype may influence GMP manufacturing of adaptive NK-cell products. These findings support prospective evaluation of donor immunophenotyping as a biomarker for donor qualification and manufacturing optimization to facilitate standardized off-the-shelf adaptive NK-cell therapies after allo-HSCT.<\/jats:p>","DOI":"10.3390\/cancers18172732","type":"journal-article","created":{"date-parts":[[2026,8,24]],"date-time":"2026-08-24T09:20:02Z","timestamp":1787563202000},"page":"2732","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Donor NKG2A\/NKG2C Immunophenotype Influences the GMP-Compliant Manufacturing Potential of NK Cells for Adoptive Immunotherapy"],"prefix":"10.3390","volume":"18","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-4376-5997","authenticated-orcid":false,"given":"Rut","family":"Meseguer","sequence":"first","affiliation":[{"name":"Hematology Service, Hospital Cl\u00ednico Universitario, Health Research Institute INCLIVA, 46010 Valencia, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1594-3648","authenticated-orcid":false,"given":"Cristobal","family":"Aguilar","sequence":"additional","affiliation":[{"name":"Advanced Therapies Unit, Health Research Institute La Fe, 46026 Valencia, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Paula","family":"Amat","sequence":"additional","affiliation":[{"name":"Hematology Service, Hospital Cl\u00ednico Universitario, Health Research Institute INCLIVA, 46010 Valencia, Spain"},{"name":"Medicine Department, University of Valencia, 46010 Valencia, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6364-8838","authenticated-orcid":false,"given":"Luis","family":"Larrea","sequence":"additional","affiliation":[{"name":"Transfusion Centre of the Valencian Community, 46014 Valencia, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ana","family":"Bonora","sequence":"additional","affiliation":[{"name":"Advanced Therapies Unit, Health Research Institute La Fe, 46026 Valencia, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Pedro","family":"Chor\u00e3o","sequence":"additional","affiliation":[{"name":"Hematology Service, Hospital Universitario y Polit\u00e9cnico La Fe, 46026 Valencia, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Francisco","family":"Boix","sequence":"additional","affiliation":[{"name":"Transfusion Centre of the Valencian Community, 46014 Valencia, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jose Luis","family":"Pi\u00f1ana","sequence":"additional","affiliation":[{"name":"Hematology Service, Hospital Cl\u00ednico Universitario, Health Research Institute INCLIVA, 46010 Valencia, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rosa","family":"Guerrero","sequence":"additional","affiliation":[{"name":"Transfusion Centre of the Valencian Community, 46014 Valencia, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3275-5593","authenticated-orcid":false,"given":"Pau","family":"Montesinos","sequence":"additional","affiliation":[{"name":"Hematology Service, Hospital Universitario y Polit\u00e9cnico La Fe, 46026 Valencia, Spain"},{"name":"Centro de Investigaci\u00f3n Biom\u00e9dica en Red de C\u00e1ncer (CIBERONC), 28029 Madrid, Spain"},{"name":"Hematology Research Group, Health Research Institute La Fe, 46026 Valencia, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bel\u00e9n","family":"Vera","sequence":"additional","affiliation":[{"name":"Transfusion Centre of the Valencian Community, 46014 Valencia, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6433-4825","authenticated-orcid":false,"given":"Dolores","family":"Planelles","sequence":"additional","affiliation":[{"name":"Transfusion Centre of the Valencian Community, 46014 Valencia, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9846-2777","authenticated-orcid":false,"given":"Mar","family":"Luis","sequence":"additional","affiliation":[{"name":"Transfusion Centre of the Valencian Community, 46014 Valencia, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1800-6198","authenticated-orcid":false,"given":"Jose Luis","family":"Poveda","sequence":"additional","affiliation":[{"name":"Pharmacy Department, Hospital Universitario y Polit\u00e9cnico La Fe, 46026 Valencia, Spain"},{"name":"Pharmacy Research Group, Health Research Institute La Fe, 46026 Valencia, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8354-768X","authenticated-orcid":false,"given":"Javier","family":"De la Rubia","sequence":"additional","affiliation":[{"name":"Hematology Service, Hospital Universitario y Polit\u00e9cnico La Fe, 46026 Valencia, Spain"},{"name":"Centro de Investigaci\u00f3n Biom\u00e9dica en Red de C\u00e1ncer (CIBERONC), 28029 Madrid, Spain"},{"name":"Hematology Research Group, Health Research Institute La Fe, 46026 Valencia, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sergi","family":"Querol","sequence":"additional","affiliation":[{"name":"Fundaci\u00f3n Internacional Josep Carreras, 08021 Barcelona, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3650-1547","authenticated-orcid":false,"given":"Cristina","family":"Arbona","sequence":"additional","affiliation":[{"name":"Transfusion Centre of the Valencian Community, 46014 Valencia, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5978-2578","authenticated-orcid":false,"given":"Manuel","family":"Guerreiro","sequence":"additional","affiliation":[{"name":"Hematology Research Group, Health Research Institute La Fe, 46026 Valencia, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-3702-0817","authenticated-orcid":false,"given":"Carlos","family":"Solano","sequence":"additional","affiliation":[{"name":"Hematology Service, Hospital Cl\u00ednico Universitario, Health Research Institute INCLIVA, 46010 Valencia, Spain"},{"name":"Medicine Department, University of Valencia, 46010 Valencia, Spain"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,8,23]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1345","DOI":"10.1182\/blood.2022016867","article-title":"Diagnosis and 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