CD94-driven in vitro expansion of highly functional adaptive NKG2C+ NKG2A- CD57+ NK cells from CMV+ healthy donors
BackgroundAdaptive human natural killer (NK) cells are an NK cell subpopulation arising upon cytomegalovirus (CMV) infection. They are characterized by CD94/NKG2C expression, a mature CD57+KIR+NKG2A– phenotype, a prolonged lifespan, and remarkable antitumor functions. In light of these features, ada...
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Frontiers Media S.A.
2025-01-01
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| Series: | Frontiers in Immunology |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fimmu.2025.1481745/full |
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| author | Chiara Giordano Simona Carlomagno Simona Carlomagno Michela Falco Claudia Cantoni Claudia Cantoni Massimo Vitale Ignazio Caruana Johannes Dirks Alberto Serio Letizia Muccio Giulia Bartalucci Alessandra Bo Franco Locatelli Franco Locatelli Cristina Bottino Cristina Bottino Simona Sivori Simona Sivori Mariella Della Chiesa Mariella Della Chiesa |
| author_facet | Chiara Giordano Simona Carlomagno Simona Carlomagno Michela Falco Claudia Cantoni Claudia Cantoni Massimo Vitale Ignazio Caruana Johannes Dirks Alberto Serio Letizia Muccio Giulia Bartalucci Alessandra Bo Franco Locatelli Franco Locatelli Cristina Bottino Cristina Bottino Simona Sivori Simona Sivori Mariella Della Chiesa Mariella Della Chiesa |
| author_sort | Chiara Giordano |
| collection | DOAJ |
| description | BackgroundAdaptive human natural killer (NK) cells are an NK cell subpopulation arising upon cytomegalovirus (CMV) infection. They are characterized by CD94/NKG2C expression, a mature CD57+KIR+NKG2A– phenotype, a prolonged lifespan, and remarkable antitumor functions. In light of these features, adaptive NK cells represent suitable candidate to design next-generation therapies, based on their enhanced effector function which could be further boosted by Chimeric Antigen Receptors-engineering, or the combination with cell engagers. For therapeutic approaches, however, it is key to generate large numbers of functional cells.PurposeWe developed a method to efficiently expand adaptive NK cells from NK-enriched cell preparations derived from the peripheral blood of selected CMV-seropositive healthy donors. The method is based on the use of an anti-CD94 monoclonal antibody (mAb) combined with IL-2 or IL-15.ResultsBy setting this method we were able to expand high numbers of NK cells showing the typical adaptive phenotype, CD94/NKG2C+ CD94/NKG2A- CD57+, and expressing a single self-inhibitory KIR. Expanded cells maintained the CMV-induced molecular signature, exhibited high ADCC capabilities and degranulation against a HLA-E+ target. Importantly, mAb-expanded adaptive NK cells did not upregulate PD-1 or other regulatory immune checkpoints that could dampen their function.ConclusionsBy this study we provide hints to improve previous expansion methods, by eliminating the use of genetically modified cells as stimulators, and obtaining effectors not expressing unwanted inhibitory receptors. This new protocol for expanding functional adaptive NK cells is safe, cost-effective and easily implementable in a GMP context, suitable for innovative immunotherapeutic purposes. |
| format | Article |
| id | doaj-art-8668255b85144187986433b0794c9b6c |
| institution | Kabale University |
| issn | 1664-3224 |
| language | English |
| publishDate | 2025-01-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Immunology |
| spelling | doaj-art-8668255b85144187986433b0794c9b6c2025-01-31T06:41:04ZengFrontiers Media S.A.Frontiers in Immunology1664-32242025-01-011610.3389/fimmu.2025.14817451481745CD94-driven in vitro expansion of highly functional adaptive NKG2C+ NKG2A- CD57+ NK cells from CMV+ healthy donorsChiara Giordano0Simona Carlomagno1Simona Carlomagno2Michela Falco3Claudia Cantoni4Claudia Cantoni5Massimo Vitale6Ignazio Caruana7Johannes Dirks8Alberto Serio9Letizia Muccio10Giulia Bartalucci11Alessandra Bo12Franco Locatelli13Franco Locatelli14Cristina Bottino15Cristina Bottino16Simona Sivori17Simona Sivori18Mariella Della Chiesa19Mariella Della Chiesa20Department of Experimental Medicine, University of Genoa, Genoa, ItalyDepartment of Experimental Medicine, University of Genoa, Genoa, ItalyDepartment of Medicine (DMED), University of Udine, Udine, ItalyDepartment of Services, IRCCS Istituto Giannina Gaslini, Genova, ItalyDepartment of Experimental Medicine, University of Genoa, Genoa, ItalyDepartment of Services, IRCCS Istituto Giannina Gaslini, Genova, ItalyU.O. Patologia e Immunologia sperimentale, IRCCS Ospedale Policlinico San Martino, Genoa, ItalyHematology, Oncology and Stem Cell Transplantation Unit, Department of Pediatrics, University Hospital Würzburg, Würzburg, GermanyDepartment of Pediatrics, University Hospital Würzburg, Würzburg, GermanyHematology and Cell Therapy, IRCCS Ospedale Policlinico San Martino, Genoa, ItalyDepartment of Experimental Medicine, University of Genoa, Genoa, ItalyHematology and Cell Therapy, IRCCS Ospedale Policlinico San Martino, Genoa, ItalyHematology and Cell Therapy, IRCCS Ospedale Policlinico San Martino, Genoa, ItalyUnit of Hematology/Oncology, Cell and Gene Therapy, Bambino Gesù Children’s Hospital, IRCCS, Rome, ItalyDepartment of Life Sciences and Public Health, Catholic University of the Sacred Heart, Rome, ItalyDepartment of Experimental Medicine, University of Genoa, Genoa, ItalyDepartment of Services, IRCCS Istituto Giannina Gaslini, Genova, ItalyDepartment of Experimental Medicine, University of Genoa, Genoa, Italy0Direzione Scientifica, IRCCS Ospedale Policlinico San Martino, Genoa, ItalyDepartment of Experimental Medicine, University of Genoa, Genoa, Italy0Direzione Scientifica, IRCCS Ospedale Policlinico San Martino, Genoa, ItalyBackgroundAdaptive human natural killer (NK) cells are an NK cell subpopulation arising upon cytomegalovirus (CMV) infection. They are characterized by CD94/NKG2C expression, a mature CD57+KIR+NKG2A– phenotype, a prolonged lifespan, and remarkable antitumor functions. In light of these features, adaptive NK cells represent suitable candidate to design next-generation therapies, based on their enhanced effector function which could be further boosted by Chimeric Antigen Receptors-engineering, or the combination with cell engagers. For therapeutic approaches, however, it is key to generate large numbers of functional cells.PurposeWe developed a method to efficiently expand adaptive NK cells from NK-enriched cell preparations derived from the peripheral blood of selected CMV-seropositive healthy donors. The method is based on the use of an anti-CD94 monoclonal antibody (mAb) combined with IL-2 or IL-15.ResultsBy setting this method we were able to expand high numbers of NK cells showing the typical adaptive phenotype, CD94/NKG2C+ CD94/NKG2A- CD57+, and expressing a single self-inhibitory KIR. Expanded cells maintained the CMV-induced molecular signature, exhibited high ADCC capabilities and degranulation against a HLA-E+ target. Importantly, mAb-expanded adaptive NK cells did not upregulate PD-1 or other regulatory immune checkpoints that could dampen their function.ConclusionsBy this study we provide hints to improve previous expansion methods, by eliminating the use of genetically modified cells as stimulators, and obtaining effectors not expressing unwanted inhibitory receptors. This new protocol for expanding functional adaptive NK cells is safe, cost-effective and easily implementable in a GMP context, suitable for innovative immunotherapeutic purposes.https://www.frontiersin.org/articles/10.3389/fimmu.2025.1481745/fulladaptive NK cellsCD94/NKG2Cexpansionmonoclonal antibodyimmunotherapyADCC |
| spellingShingle | Chiara Giordano Simona Carlomagno Simona Carlomagno Michela Falco Claudia Cantoni Claudia Cantoni Massimo Vitale Ignazio Caruana Johannes Dirks Alberto Serio Letizia Muccio Giulia Bartalucci Alessandra Bo Franco Locatelli Franco Locatelli Cristina Bottino Cristina Bottino Simona Sivori Simona Sivori Mariella Della Chiesa Mariella Della Chiesa CD94-driven in vitro expansion of highly functional adaptive NKG2C+ NKG2A- CD57+ NK cells from CMV+ healthy donors Frontiers in Immunology adaptive NK cells CD94/NKG2C expansion monoclonal antibody immunotherapy ADCC |
| title | CD94-driven in vitro expansion of highly functional adaptive NKG2C+ NKG2A- CD57+ NK cells from CMV+ healthy donors |
| title_full | CD94-driven in vitro expansion of highly functional adaptive NKG2C+ NKG2A- CD57+ NK cells from CMV+ healthy donors |
| title_fullStr | CD94-driven in vitro expansion of highly functional adaptive NKG2C+ NKG2A- CD57+ NK cells from CMV+ healthy donors |
| title_full_unstemmed | CD94-driven in vitro expansion of highly functional adaptive NKG2C+ NKG2A- CD57+ NK cells from CMV+ healthy donors |
| title_short | CD94-driven in vitro expansion of highly functional adaptive NKG2C+ NKG2A- CD57+ NK cells from CMV+ healthy donors |
| title_sort | cd94 driven in vitro expansion of highly functional adaptive nkg2c nkg2a cd57 nk cells from cmv healthy donors |
| topic | adaptive NK cells CD94/NKG2C expansion monoclonal antibody immunotherapy ADCC |
| url | https://www.frontiersin.org/articles/10.3389/fimmu.2025.1481745/full |
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