Fig. 4: CD8⁺ exhausted T cells express Ccl4.

A Schematic representation of the optic glioma immune circuit. Created in BioRender. Chatterjee, J. (2024) BioRender.com/e28x983. B Ccl4 expression in the dura, optic nerve (ON), cerebellum (CBL), brainstem (BSTM), cortex (CTX), hippocampus (HC), deep cervical lymph nodes (dCLN), and superficial cervical lymph nodes (CLN) of 12-week-old Nf1-OPG mice, following normalization to the optic nerve. Data are represented as mean ± SD. (dura, n = 6 mice; ON, n = 3, 2 pooled optic nerves per sample; CBL, n = 6 mice; BSTM, n = 5 mice; CTX, n = 5 mice; HC, n = 6 mice; dCLN, n = 6 mice; CLN, n = 6 mice). A one-way ANOVA test was performed followed by a Holm-Šídák post-test correction. Exact P values are included. C Dot plot showing Ccl4 expression from 12-week-old Nf1-OPG mouse optic nerves (corresponds to the dataset shown in Fig. 2; n = 10 pooled mice), meningeal dura (corresponds to the dataset shown in Fig. 3D; n = 10 pooled mice), and PBMC (corresponds to the dataset shown in Fig. 3B; n = 10 pooled mice) scRNAseq datasets. D Volcano plot showing fold change and P value comparing CD8⁺ exhausted T cells and the other T cell populations in the optic nerves of 12-week-old Nf1-OPG mice (corresponds to the dataset shown in Fig. 2; n = 10 pooled mice). Upregulated genes in red, downregulated genes in blue. Differential analyses were performed using gene specific analysis (GSA). E Volcano plot showing fold change and P value comparing CD8⁺ exhausted T cells in the optic nerves (corresponds to the dataset shown in Fig. 2; n = 10 pooled mice) and PMBC T cells of 12-week-old Nf1-OPG mice (corresponds to the dataset shown in Fig. 3B; n = 10 pooled mice). Upregulated genes in red, downregulated genes in blue. Differential analyses were performed using gene specific analysis (GSA). F Volcano plot showing fold change and P value comparing CD8⁺ exhausted T cells in the optic nerves (corresponds to the dataset shown in Fig. 2, Supplementary Fig. 2; n = 10 pooled mice in each dataset) and T cells from the dura of 12-week-old Nf1-OPG mice (corresponds to the dataset shown in Fig. 3D; n = 10 pooled mice). Upregulated genes in red, downregulated genes in blue. Differential analyses were performed using gene specific analysis (GSA). G Volcano plot showing fold change and P value comparing CD8⁺ TEX in the optic nerves (corresponds to the dataset shown in Fig. 2; n = 10 pooled mice, and to the dataset shown in Supplementary Fig. 2; n = 10 pooled mice) and CD8⁺ non-TEX T cells of 12-week-old Nf1-OPG mice (corresponds to the dataset shown in Fig. 2; n = 10 pooled mice, and to the dataset shown in Supplementary Fig. 2; n = 10 pooled mice). Upregulated genes in red, downregulated genes in blue. Differential analyses were performed using gene specific analysis (GSA). H Graph showing levels of secreted Ccl4 in the TCM by ELISA of unstimulated and CD8⁺ exhausted T cells (α-CD3+α-CD28 antibody stimulated) with and without FK506 treatment (none, n = 4; α-CD3+α-CD28, n = 4; α-CD3+α-CD28 + FK506, n = 4). A one-way ANOVA test was performed followed by a Holm-Šídák post-test correction. Data are represented as mean ± SD. Exact P values are indicated.