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. 2023 Sep 8;14:5546. doi: 10.1038/s41467-023-41338-y

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© The Author(s) 2023

Open Access This 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 — a Cytotoxicity of target cells by CD3⁺ nfP2X7-M-CAR-T cells compared with donor-matched untransduced (UT) cells generated in parallel. Target cell lines include: DU145 (prostate cancer), PC3 (prostate cancer), BT-549 (breast cancer), MDA-MB-231 (breast cancer), PC9 (lung cancer), NC1-H460 (lung cancer), C32 (melanoma), Sk-Mel-28 (melanoma), Sk-Mel-5 (melanoma), Namalwa (lymphoma), Ramos (lymphoma), Raji (lymphoma), SH-SY5Y (neuroblastoma), Be(2)-M17 (neuroblastoma), Kelly (neuroblastoma), SK-ND-Z (neuroblastoma), UM-SCC-1 (head and neck carcinoma), RD (rhabdomyosarcoma), LIM1215 (colorectal cancer), HCT116 (colorectal cancer), AsPC-1 (pancreatic cancer), OVCAR5 (ovarian cancer), OVCAR3 (ovarian cancer), RPMI-8226 (myeloma). CAR-T cells were co-cultured with target cancer cell lines at effector:target E:T ratios of 10:1, 3:1 and 1:1 for 16 h. Specific cytotoxicity was measured using a BrightGlo luciferase-based cytotoxicity assay system; paired T test comparing the nfP2X7-M CAR-T with CD3⁺ UT at the indicated time points. Pooled data from three independent experiments. Statistical significance was evaluated by paired T test. Exact p values on the graph, ns: non-significant. b Table listing cancer types and cancer cell lines tested. Cytotoxicity values at E:T ratio of 10:1, with cytotoxicity values from untransduced control T cells subtracted. c Representative graph showing changes in normalised cell index over time (in hours) for the human prostate cancer cell line PC3 when untreated (black) or treated with either untransduced T cells (UT) (dark blue) or nfP2X7-M CAR T cells (orange). Black vertical line reflects time point at which T cells were added. d The area under the curves were measured and normalised to the cancer cell alone control. Data represents mean ± SEM. Data is pooled from independent experiments for MDA-MB-231 (n = 6), PC3 (n = 6) and OVCAR3 (n = 4) target cell lines, where each dot represents an experiment conducted with T cells from a separate donor. Statistical significance was evaluated by paired T test. Exact p values on the graph, ns: non-significant.