We developed an innovative and efficient, feeder-free culture method to genetically modify and expand peripheral blood-derived NK cells with high proliferative capacity, while preserving the responsiveness of their native activating receptors. Activated peripheral blood NK cells were efficiently transduced by a retroviral vector, carrying a second-generation CAR targeting CD19. CAR expression was demonstrated across the different NK-cell subsets. CAR.CD19-NK cells display higher antileukemic activity toward CD19+ cell lines and primary blasts obtained from patients with B-cell precursor ALL compared with unmodified NK cells. In vivo animal model data showed that the antileukemia activity of CAR.CD19-NK cell is superimposable to that of CAR-T cells, with a lower xenograft toxicity profile. These data support the feasibility of generating feeder-free expanded, genetically modified peripheral blood NK cells for effective “off-the-shelf” immuno-gene-therapy, while their innate alloreactivity can be safely harnessed to potentiate allogeneic cell therapy.
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A heartfelt thanks for the helpful suggestions to Alessandro Moretta who passed away before the final acceptance. Supported by grants awarded by Fondazione AIRC per la Ricerca sul Cancro-Special Project 5 × 1000 no. 9962 (FL and LM), AIRC IG 2018 id. 21724 (FL), AIRC IG 2017 Id. 15704 (SS), AIRC IG 2017 Id. 15283 (LM), Ricerca Finalizzata GR-2013-02359212 (CQ), Ministero dell’Università e della Ricerca (grant PRIN 2017WC8499_004 to FL and SS) Ricerca Corrente (FL, CQ, and BDA).
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Quintarelli, C., Sivori, S., Caruso, S. et al. Efficacy of third-party chimeric antigen receptor modified peripheral blood natural killer cells for adoptive cell therapy of B-cell precursor acute lymphoblastic leukemia. Leukemia 34, 1102–1115 (2020). https://doi.org/10.1038/s41375-019-0613-7
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