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. 2019 Apr 30;10:957. doi: 10.3389/fimmu.2019.00957

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Copyright © 2019 Ingegnere, Mariotti, Pelosi, Quintarelli, De Angelis, Tumino, Besi, Cantoni, Locatelli, Vacca and Moretta.

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) and the copyright owner(s) 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.

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Electroporation of a functional CAR-antiCD19 transgene in NK cells increases their cytolytic activity. (A) Schematic representation of the different domains of the two CAR constructs. (B) Representative gating strategy used to test the electroporation efficiency of I and II generation CAR constructs in the different NK cell populations. (C) Electroporation efficiency of CD19⁺CAR plasmid in untransfected, mock-transfected and CAR-transfected activated NK cells. CAR expression was evaluated on DAPI-negative live cells using the delta (Δ) CD34 marker. Cytometric profiles of unstained (empty profile), isotype stained (gray profile) and anti-ΔCD34 mAb stained (black profile) NK cells are shown. (D) Percentage of cytotoxicity was evaluated by flow cytometry on propidium iodide (PI) positive target cells. CD19⁻ cell lines (upper panel) and CD19⁺ (lower panel) were used as target cells. Effector: Target (E:T) ratios are indicated. A representative experiment out of 3 performed is shown.