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. 2022 Nov 2;12:18509. doi: 10.1038/s41598-022-23016-z

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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 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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SC79 increases tumor infiltration of CD8-T cells and CD8 T-cells expressing IFNγ. (a) Profiles showing an increase in the per cent of CD8alpha x Tbet expression in responsive mice (left panel). Histogram showing an increase in numbers of CD8alpha x Tbet TILs responder mice (right panel) (n = 3). (b) Profiles show an increase in the per cent of CD8alpha x PD-1 expression (left panel). Histogram showing the increase in numbers of CD8alpha x PD-1 TILs (right panel) (n = 3). (c) FACS profiles show an increase in the per cent of Tbet x PD-1 expression (left panel). Histogram showing the increase in numbers of Tbet x PD-1 TILs (right panel). (d) Profiles showing an increase in the per cent of pSTAT5 x Tbet expression (left panel). Histogram showing the increase in numbers of pSTAT5 x Tbet 1 TILs (right panel) (n = 3). (e) viSNE analysis of TILs with a focus on CD8+ T-cells (circled). Subpopulations of CD8 T-cells were obtained by the expression of Tbet, PD-1, and CD73 (n = 3). (f) Histogram showing the frequency and cell number of CD8 T-cell subpopulations outlined in Fig. 2e. (g) Profiles showing the frequency of CD8 T-cell TILs expressing IFN-γ. (h) Histogram showing the increase in the percentage of CD8 T-cells expressing IFN-γ from CTL (n = 4), SC79 R (n = 3) and SC79 NR (n = 3) mice. An increase in CD8+ IFN-γ+ TILs was seen in responder but not non-responder mice. i, IFN-γ expression on CD8 T-cells is increased by treating ex vivo TILs with SC79 from control mice. TILs from untreated mice were cultivated in a 96-well plate for 3 days with anti-CD3 with recombinant IL-2 in the presence or absence of SC79. While 54.6 per cent of CD8+ control cells showed IFN-γ expression, this increased further to 78.9 per cent due to culturing in SC79 (n = 3). j, IFN-γ expression in CD8 T-cell from a healthy donor after 48 h of culture with or without SC79 (n = 4). Results from the figure (a) to (g) were obtained after CyTOF acquisition from a pool of 3 tumors of each group. For statistical analyses, one-way ANOVA and a non-parametric unpaired t-test were used. *p < 0.05, **p < 0.01. The R package CATALYST v1.12.1 was used to perform unsupervised clustering, and generate the heatmaps and tSNE plots. https://bioconductor.org/packages/release/bioc/html/CATALYST.html. https://f1000research.com/articles/6-748.