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. 2023 Apr 13;72(8):2573–2583. doi: 10.1007/s00262-023-03443-1

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© The Author(s) 2023

Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 3 — Cytotoxic activity of knockout CD19-CAR-NK92 cells versus CD19⁺ target cells. A Specific lysis (%) of Daudi (top) and Karpas422 cells (bottom) by wildtype (white), reference (green), FasL-ko (blue), Prf1-ko (red) and simultaneous FasL- & Prf1-ko CD19-CAR-NK92 cells (yellow) at E:T ratios of 1:4, 1:1 and 4:1. Cells were co-incubated for 4 h. Two cell clones of each knockout type were tested in triplicates and results were combined into one data group. Values are expressed as means of multiple determination ± SEM. P values were calculated by one-way ANOVA followed by Tukey’s test. B Proportion (%) of perforin/granzyme pathway, FasL/FasR pathway and others of the total cytotoxic efficacy of CD19-CAR-NK92 cells against Daudi (top) and Karpas422 cells (bottom) at E:T ratios of 1:4, 1:1 and 4:1. The proportions were calculated as following: % = average reduction in cytotoxicity due to respective knockout/cytotoxicity of reference effector cells. The residual lysis of target cells by double-ko effector cells was used to calculate the influence of other pathways, non-perforin and non-FasL mediated