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. 2024 Aug 13;15:6949. doi: 10.1038/s41467-024-51025-1

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© The Author(s) 2024

Open Access This article is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License, which permits any non-commercial use, sharing, 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 you modified the licensed material. You do not have permission under this licence to share adapted material derived from this article or parts of it. 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-nc-nd/4.0/.

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Fig. 4 — Flow cytometry analysis of intracellular cytokine staining of Vδ1 + and Vδ2 + T cells in HC or CRC. a Overlay of representative histograms and corresponding bar plots of tested cytokines by flow cytometric analysis in Vδ1 + and Vδ2 + T cells in HC (light cyan) or CRC (light pink) and stimulated (PMA/iono) HC (cyan) or CRC (pink, n = 16 per group). In CRC, Vδ1 + display reduced expression of TRAIL, TNF-α and IFN-γ which is reversible upon stimulation. b Percentage of Ki-67 + Vδ1 + , Vδ2 + and TCRγδ negative T cells in HC (white) compared to CRC (gray; n = 8 per group). Increased expression of Ki-67 in Vδ1 + and TCRγδ negative T cells in CRC indicate in tissue proliferation in contrast to the Ki-67 negative Vδ2 + T cells which potentially influx from the periphery. Overlay of representative histograms of Ki-67 expression in HC (cyan) and CRC (pink). c Increased Ki-67+ expression of Vδ1 + T cells isolated of CRC was mirrored in elevated proliferation (gray) during IL-2 alone or CD3/CD28 stimulation as compared to HC (white; n = 3 per group). d Pairwise comparison of killing assay with Vδ1 + and Vδ2 + T cells with two different CRC lines (HT29 in black, SW480 in blue; n = 14 per group). e Percentage of dead HT29 after exclusion of baseline HT29 cell death in a killing assay with Vδ1 + and Vδ2 + T cells isolated from HC (cyan) and CRC (pink). Two conditions were tested: Vδ1 + and Vδ2 + T cells with (cyan, pink) and without PMA/iono stimulation (light cyan, light pink; n = 5 per group). f CD107a expression of Vδ1 + and Vδ2 + T cells from HC (cyan) and CRC (pink) after killing of HT29 as in (e). g Representative FACS plot of dead HT29 in a killing assay with Vδ1 + T cells with no stimulation and stimulated with PMA/ionomycin. Data points and error bars represent the mean ± SD (a−f); one-way ANOVA with Fisher’s LSD (a−f); Ctrl; Control; p values are shown on the graphs.