Fig. 5

CPR treatment stimulates T-cell proliferation, activation, and improves lymphoid effector-to-suppressor ratios. Mice bearing established mEER tumors were harvested after 1 week of treatment (day 23) for assessment of lymphoid cellular changes using flow cytometry both in tdLN (a and b) and tumor (c-f; see Additional file 12: Figure S12 for lymphoid gating strategies). a Percentage of lymphoid subsets within the tdLN (among CD45⁺ cells; Dunn’s multiple comparison test; N = 2; n = 7–12 per group). b Aggregate flow cytometry scatterplots showing Ki67 expression among CD8⁺ T cells within the tdLN (percentages show mean ⁺/− SD; N = 1 representative of 2; n = 6 aggregate samples per group). c Pie-chart showing average tumor-infiltrating lymphoid and myeloid subsets as a fraction of total CD45⁺ cells for CPR treated tumors at days 23, 33, and 37 (N = 2; n = 10–16/group). d Aggregate flow cytometry scatter plots of CPR treated tumors showing CD8⁺ T cells (top panels), CD4⁺ T cells and regulatory T cells (bottom panels) at each day of treatment progression (percentages show mean ⁺/− SD; N = 1, representative of 2; n = 6 aggregate samples per day). e Summary of CPR intratumoral CD8⁺ and regulatory T cell percentages (among CD45⁺ cells; left y-axis) and the ratio of CD8⁺ T cell / regulatory T cells (right y-axis) at days 23, 33, and 37 of treatment (N = 2; n = 10–16/group). f Intratumoral CD8⁺ T cell phenotypic marker expression at days 23, 33, and 37 of CPR treatment progression. Data is represented as a z-score of the phenotypic marker’s median fluorescence intensity (MFI) compared to size-matched control tumors (N = 2; n = 11–13 per group). *p < 0.05; **p < 0.01; ***p < 0.001