Figure 3: Exercise-induced tumor protection requires CD8 T cells.

(a-f) WT mice were injected orthotopically and exercise was started at Day 1. a. Day 21 FFPE tumor sections were stained by IF for CD8, CD3, and DAPI. The % of CD8+ CD3+ cells was quantified. Each dot = one mouse; n=4. (b-d) scRNAseq of Day 21 tumors: b. KEGG analysis of Cluster 11 CD8 T cells. c. Canonical pathway perturbations in IPA (Ingenuity Pathway Analysis) in exercise CD8 T cells. d. Relative mRNA expression of select markers on CD8 T cells, all FDR p-adjusted values < 0.05. e. FC for GZMB, T-bet, CD44, ICOS, and PD-1 as a fraction of CD8. Each dot = one mouse (n = 5–9). Experiment was repeated ten times. f. Control and exercise mice were treated with IgG or anti-CD8. Day 21 tumor weights are shown, each dot = one mouse (n=7–8). Experiment was repeated three times. g. IHC for GZMB+ and CD8+ cells on FFPE tumor sections from exercised pancreatic cancer patients or historical controls; positive cells/FOV quantified. Each dot = one patient (n = 9). h-i. Kaplan-Meier survival curve of exercise patients stratified by CD8 high [top 50%] (n =5), and CD8 low [bottom 50%] (n = 4) (h), or GZMB high (n =5), GZMB low (n = 4) (i), based on levels in (g). j. ELISA for epi on sera isolated 20 minutes after completion of exercise from WT mice. Each dot = one mouse (n = 5), experiment was repeated twice. (k-l) WT mice were injected orthotopically and exercise was started at Day 1. k. At Day 21, CD3+ T cells in tumor and blood were assessed by FC 20 minutes post-exercise. Each dot = one mouse (n= 7–9). Representative contour plots are shown. Experiment was repeated three times. (l) Mice were treated with PBS or FTY720. Day 21 tumor weights are shown, each dot = one tumor (n =7–8). Experiment was repeated three times. (p > 0.05 = ns, p < 0.05 = *, p < 0.01 = **, p < 0.001 = ***). See Supp. 3–6