Fig. 5: Induction of CAF senescence reduces rates of activated CD8+ T cells. | Nature Communications

Fig. 5: Induction of CAF senescence reduces rates of activated CD8+ T cells.

From: Senescent cancer-associated fibroblasts in pancreatic adenocarcinoma restrict CD8+ T cell activation and limit responsiveness to immunotherapy in mice

Fig. 5

a Diagram of experimental procedure: mScarlet-expressing mouse CAFs were co-injected into mouse pancreata with 6555c3 KPC tumor cells. Senescence was induced in CAFs in half of the mice through tamoxifen (TAM) treatment one and three days later, and tissues were collected one week after cell transplantation. b Lesion sections from untreated (-TAM) and tamoxifen-treated ( + TAM) mice, further analyzed in panels c-g, 1 week after co-injection, stained for mScarlet marking the injected CAFs, and for CK8 marking the co-injected KPC cells. c Lesion sections from control and tamoxifen-treated mice stained for CD8. d Quantification of CD8+ T cells in lesions from untreated and tamoxifen treated mice. Values were measured by image analysis and are presented relative to mean of control mice. n = 4 tumors. e Lesion sections from untreated and tamoxifen-treated mice co-stained for CD8 (green) and Gzmb (red). f Percentages of Gzmb+ cells out of CD8+ T cells in lesions from untreated and tamoxifen-treated mice, scored by image analysis. n = 5 tumors. g Quantification of total T cells (CD3+), regulatory T cells (FoxP3+), and monocytes (CD11b+) in same lesions. Values were measured by image analysis and are presented relative to mean of control mice. n = 5 tumors. h Percentage of IFNγ+ and GZMB+ human CD8+ T cells following treatment with conditioned media from non-senescent (Non-sen) or senescent (Sen) human PDAC CAFs, measured by FACS. n = 8 replicates from two experiments. i IFNγ levels secreted by human T cells following treatment with conditioned media from non-senescent or senescent human CAFs, measured by ELISA. n = 4 replicates. j Percentage IFNγ+ Gzmb+ mouse CD8+ T cells following treatment with conditioned media from non-senescent or senescent mouse CAFs, measured by FACS. Where indicated, CAFs were pretreated with IFNγ for 48 h prior to conditioned media collection. n = 3 replicates. All values represent mean ± SEM. Pairwise two-tailed t tests compare senescence to non-senescence in all graphs. Scale bars = 20 μm.

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