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. 2023 Mar 16;24(4):612–624. doi: 10.1038/s41590-023-01448-7

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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 license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license 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 license, visit http://creativecommons.org/licenses/by/4.0/.

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Extended Data Fig. 7 — a. Diagram showing the workflow to obtain pure tumor cell, αβ or γδ T cell populations from RCC biopsies and conditions for the autologous killing assay co-cultures. b. Flow cytometry gating strategy used to sort TIL populations from digested RCC tumor biopsies. c. Flow cytometry dot plots showing the sorted αβ CD8+ or γδ TIL populations after downstream expansion before adding them to cytotoxicity assays in co-culture with target autologous tumor cells. d. Flow cytometry contour plots showing expression of PD-1, TIM3 and 4-1BB in expanded Vδ2- T cells used in autologous killing assays. e. Diagram showing the workflow for generating fresh autologous TIL:tumor cocultures for immune stimulation and readout by ELISpot. Pseudocolor flow cytometry plots show gating strategy for cell sorting of target populations. f. Human IFNγ ELISpot representative individual well images from TIL:tumor cocultures with QC-passed counts shown on the upper left edge of each well. Row titles stand for stimulation conditions. Column titles show sorted cell populations present in the wells. Created with BioRender.com.