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BRIEF RESEARCH REPORT article

Front. Immunol.
Sec. Cancer Immunity and Immunotherapy
Volume 16 - 2025 | doi: 10.3389/fimmu.2025.1481745
This article is part of the Research Topic Community Series in New Insights into Innate Immune Cell-Based Immunotherapies in Cancer: Volume II View all articles

CD94-driven in vitro expansion of highly functional adaptive NKG2C + NKG2A -CD57 + NK cells from CMV + healthy donors

Provisionally accepted
Chiara Giordano Chiara Giordano 1Simona Carlomagno Simona Carlomagno 2Michela Falco Michela Falco 3Claudia Cantoni Claudia Cantoni 1,3Massimo Vitale Massimo Vitale 4Ignazio Caruana Ignazio Caruana 5Johannes Dirks Johannes Dirks 6Letizia Muccio Letizia Muccio 1Franco Locatelli Franco Locatelli 7,8Cristina Bottino Cristina Bottino 1,3Simona Sivori Simona Sivori 1,4Mariella Della Chiesa Mariella Della Chiesa 1,4*
  • 1 Department of Experimental Medicine, School of Medical and Pharmaceutical Sciences, University of Genoa, Genova, Italy
  • 2 Department of Medicine (DMED), University of Udine, Udine, Italy, Udine, Italy
  • 3 Services Department, Laboratory of Clinical and Experimental Immunology, Giannina Gaslini Institute (IRCCS), Genoa, Italy
  • 4 Ospedale Policlinico San Martino, Genova, Italy
  • 5 Department of Pediatrics, Hematology, Oncology and Stem Cell Transplantation Unit, University Hospital Würzburg, Würzburg, Germany, Wurzburg, Germany
  • 6 Department of Pediatrics, University Hospital Würzburg, Würzburg, Bavaria, Germany
  • 7 Unit of Oncohematology, Department of Oncohematology, Cell Therapy, Gene Therapies and Hematopoietic Transplantation, Bambino Gesù Children's Hospital (IRCCS), Rome, Lazio, Italy
  • 8 Department of Life Sciences and Public Health, Catholic University of the Sacred Heart, Rome, Lazio, Italy

The final, formatted version of the article will be published soon.

    BBackground Adaptive 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. Purpose We 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. Results By 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. Conclusions By 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.

    Keywords: Adaptive NK cells, CD94/NKG2C, expansion, Monoclonal antibody, Immunotherapy, ADCC, KIR, CMV

    Received: 16 Aug 2024; Accepted: 14 Jan 2025.

    Copyright: © 2025 Giordano, Carlomagno, Falco, Cantoni, Vitale, Caruana, Dirks, Muccio, Locatelli, Bottino, Sivori and Della Chiesa. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

    * Correspondence: Mariella Della Chiesa, Department of Experimental Medicine, School of Medical and Pharmaceutical Sciences, University of Genoa, Genova, 16132, Italy

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