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. 2022 Sep 28;610(7930):161–172. doi: 10.1038/s41586-022-05192-0

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© The Author(s) 2022

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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Fig. 4 — Immuno-pharmacodynamic study on the effect of the different therapies, given twice, on the abundance, phenotype, effector function and molecular signature of intratumoural CD8⁺ T cells obtained from syngeneic mice bearing subcutaneous Panc02-H7-Fluc tumours. a, Number of intratumoural CD8⁺ T cells. b, CD8⁺ T cell to T_reg ratio within the tumour. c, Number of stem-like (PD-1⁺TCF-1⁺) CD8⁺ T cells. In a–c, n = 4 (box plots represent the median, minimum/maximum and individual points; treatment groups appear in the same order in each panel). d, Representative contour plots depicting granzyme B and TIM-3 expression on PD-1⁺TCF-1⁻CD8⁺ TILs from tumour single-cell suspensions acquired by flow cytometry 3 d after administration of the second dose of the treatment as indicated. e,f, Frequencies of granzyme B⁺TIM-3⁻ (e) and granzyme B⁻TIM-3⁺ (f) intratumoural PD-1⁺TCF-1⁻CD8⁺ T cells (n = 4; box plots represent the median, minimum/maximum and individual points). g,h, Two-dimensional (2D) UMAP visualization of CD8⁺ TILs coloured according to subset (g) and specific treatment effect (h). i, Average relative expression of selected genes (RNA and/or protein level) across the distinct T cell subsets within the CD8⁺ TILs depicted in g and h. j, Expression of selected markers, signature scores and TCR clonal expansion among CD8⁺ TILs using a 2D UMAP visualization as in g and h. log(cp10k), natural logarithm of counts per 10,000; log(clone size), natural logarithm of clone size. k, Percentage of better effectors and exhausted CD8⁺ T cells relative to all CD8⁺ TILs across the different treatments and average signature enrichment scores among effector CD8⁺ T cells per treatment group and individual animal (3–4 mice per group; box plots represent the median, minimum/maximum and individual points). In a and k, n = 3–4 mice per group per experiment, >3 independent experiments; statistical comparisons were performed using one-way or two-way ANOVA with Dunnett’s multiple-comparisons test.

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