Journal: Cell Reports Medicine
Article Title: Pan-cancer mapping of single CD8 + T cell profiles reveals a TCF1:CXCR6 axis regulating CD28 co-stimulation and anti-tumor immunity
doi: 10.1016/j.xcrm.2024.101640
Figure Lengend Snippet: CXCR6 expression increases upon ICB and is repressed by TCF1 (A and B) Left: tumor area (y axis, mean ± SEM) over time (x axis) of Mc38Ova hi (A) or B16Ova (B) implanted in wild-type (WT) mice and treated with anti-PD1 (A), anti-PD-L1 + anti-TIM3 (B), or isotype control. Right: frequency of CXCR6 + cells (y axis, mean ± SEM) in the indicated subsets (x axis) of CD8 + TILs harvested from tumors treated as above (A: n = 4–6 per group, 1 experiment. B: n = 3–4 per group, 1 experiment). (C) Expression (row-normalized TPM, color bar) of top differentially expressed genes (rows) between WT OTI (E8i-Cre - , Tcf7 FL/FL ) and Tcf7 cKO OTI (E8i-Cre + , Tcf7 FL/FL ) cells at different time points after T cell activation (columns) ( n = 3, 1 experiment). (D) Mean expression (color bar) and fraction of expressing cells (dot size) of key genes (rows) in different CD8 + T cell clusters (columns) as determined by scRNA-seq of cells from B16Ova tumors implanted in WT or Tcf7 cKO mice ( n = 3 per group combined, 1 experiment). (E) Frequency of CXCR6 + cells (y axis, mean ± SEM) in the indicated subsets (x axis) of CD8 + TILs from B16Ova tumors from WT (black) or Tcf7 cKO (green) mice ( n = 3–7 per group, representative of 2 experiments). (F) Percent input (y axis, mean ± SEM) following chromatin immunoprecipitation (ChIP) PCR of the Cxcr6 locus with anti-TCF1 or rabbit IgG control antibodies (x axis) in WT or Tcf7 cKO CD8 + T cells ( n = 8, 5 experiments combined). (G–I) Luciferase activity (RLU, relative light unit, y axis, mean ± SEM) in HEK293T cells transfected with Cxcr6 locus-containing pGL4.10 luciferase reporters together with either empty vector (control) or vectors encoding the indicated transcription factors (x axis). Firefly luciferase activity is presented relative to constitutive Renilla luciferase activity ( n = 3, representative of 2 experiments). Statistical significance was determined by linear mixed model (A left, B left), Student’s unpaired t test (A right, B right, E, F comparing WT to Tcf7 cKO anti-TCF1 samples), Student’s paired t test (F comparing WT samples), or one-way ANOVA with Tukey’s multiple comparisons test (G, H, I). NS = not significant, ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001.
Article Snippet: Anti-Mouse Tim3 APC Clone 5D12 , BD , Cat:# 567164 RRID: AB_2916481.
Techniques: Expressing, Control, Activation Assay, Chromatin Immunoprecipitation, Luciferase, Activity Assay, Transfection, Plasmid Preparation
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