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. 2024 May 15;630(8016):457–465. doi: 10.1038/s41586-024-07443-8

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© The Author(s) 2024

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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Extended Data Fig. 2 — (a) CD19.BBζ-CAR ± CV-1 (Fc-dead) treated Nalm6 tumour survival. n = 5 mice/arm. (b) Images of Nalm6 BLI on day 11 of high-dose CD19.BBζ-CAR ± CV-1 treated mice, treated as in (a). (c) Low-dose CD19.28ζ-CAR ± CV-1 (Fc-dead) treated Nalm6 tumour growth. Mean ± SEM of n = 5 mice/arm. (d) Images of Nalm6 BLI of low-dose CD19.28ζ-CAR ± CV-1 treated mice, treated as in (c). (e - f) Images (e) and quantification (f) of T cell BLI post CV-1 treatment of Nalm6 tumour-bearing mice. Top: high-dose CD19.BBζ-CAR ± CV-1, treated as in (a), on day 9 (four days post CV-1 treatment). Bottom: low-dose CD19.28ζ-CAR ± CV-1, treated as in (c), day 11 (six days post CV-1 treatment). For (f): mean ± SD of n = 5 mice for each dose condition. (g) CRISPR/Cas9 mediated CD47 knock-out (CD47_KO) efficiency in primary human T cells by flow cytometry. CD47_WT cells are CD47_KO with exogenous expression of wild-type CD47. Representative of n > 3 donors. (h) Images of Nalm6 tumour progression (top five rows) and T cells on day 11 (bottom row) by BLI in the Nalm6 – 47_KO-CD19.28ζ-CAR T model. (i) Survival of Nalm6 bearing mice shown in (h). n = 5 mice per treatment arm. (j) Representative flow cytometry plots of hCD45⁺ T cells identified in the blood of 47_WT- or 47_KO-CD19.28ζ-nLuc-CAR ± B6H12 treated, non-tumour bearing mice. (k) Images of T cell BLI following treatment with 47_WT- or 47_KO-CD19.28ζ-nLuc-CAR T cells before and after B6H12 treatment. (l) T cell BLI of mice shown in (k) treated with 47_WT- or 47_KO-CD19.28ζ-nLuc-CAR T, before and after B6H12 treatment. Dashed line indicates limit of detection. Mean ± SD of n = 5 mice. (m) hCD8⁺ and hCD4⁺ T cells in the blood on day 6 or 7 by flow cytometry following treatment 47_WT- or 47_KO-CD19.28ζ-nLuc-CAR T ± B6H12 in separate experiments, treated as in (k). Mean ± SD of n = 5 mice. [(c), (l), (m)] Two-way analysis of variance (ANOVA) test with Tukey’s multiple comparison test. ns = not significant. [(f - top)] Two-tailed Mann-Whitney test. [(f - bottom)] Unpaired two-tailed Student’s t test.

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