Extended Data Fig. 7. CD8 T cells acquire a polyfunctional effector profile upon muPD1-IL2v and are critical for its efficacy. Frequency and amount of PD-1 and IL-2Rβ per T cell in the tumor and blood of huPD1-transgenic mice.
a-b. Left, representative FACS contour plot of PD-1+ CD8 TILs secreting granzyme B, IFN-γ and TNF-α across different treatment groups; right, frequency of PD-1+ granzyme B+ and IFN-γ+ TNF-α+ CD8 TILs (n = 4 mice per group per experiment from 3 independent experiments, box plots representing median, minimum/maximum and individual points). Statistical comparisons were performed using one-way ANOVA with Tukey’s multiple comparison test. c. Tumor growth inhibition and d. CD8 T cell count in blood of syngeneic mice bearing subcutaneous Panc02-H7-Fluc tumors with or without CD8 depletion before the start of the indicated treatments (n = 11 mice per treatment group, mean ± SEM). e–h. Frequencies of receptor positive T cells and quantification of PD-1 receptors and IL-2Rβ on T cells isolated from tumors and blood of untreated human PD-1 transgenic mice bearing Panc02-H7-Fluc (n = 4 and n = 9 mice respectively, box plots representing median, minimum/maximum and individual points). i. (Top) Percentage of directly conjugated Alexa Fluor-647 parental anti-PD-1 bound to 3 days activated CD4 T cells previously exposed to increasing concentrations of either PD1-IL2v, pembrolizumab or non-blocking PD1-IL2v; (bottom) percentage of PD-1 receptors occupied by either PD1-IL2v, pembrolizumab or non-blocking PD1-IL2v and therefore unavailable for binding of the directly conjugated Alexa Fluor-647 parental anti-PD1 (n = 2 healthy donors from 2 independent experiments, mean ± SEM).