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. 2024 Jan 15;15:519. doi: 10.1038/s41467-024-44786-2

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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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Fig. 5 — a Schematic of the timeline of cytokine optimization and functional analysis. b Live total T cell counts during in vitro expansion of hCAR-Vβ2 T cells from three different donors, determined by trypan blue cell counting. c Jurkat-TRBV20-1 cell killing % and (d) live CD3- hCAR-Vβ2 T cell counts in acute repeated Jurkat-TRBV20-1 killing assay, determined by trypan blue cell counting and flow cytometry. e CD45RA+ CD45RO+ % of CD8 + CD3- hCAR-Vβ2 T cells 4 days post repeated Jurkat-TRBV20-1 cell killing assay, determined by flow cytometry. b–e N = 3 replicates in each group. Data are presented as mean values +/− SEM. f relative Jurkat-TRBV20-1 killing by chronically stimulated hCAR-Vβ2 T cells from two healthy donors generated in seven cytokine conditions, with each killing round normalized to the first round. g summary of optimized cytokine combinations for allogeneic hCAR-Vβ2 T cell expansion in vitro. Illustrations in Fig. 5a, g were created using Adobe Illustrator. All replicates are independent samples. Source data are provided as a Source Data file.