Fig. 4: ATRi/RT and anti-NKG2A/PD-L1 immunotherapy induce the proliferation of PD-1⁺ effector memory cytotoxic CD8 and CD4 T-cells in the tumour microenvironment.

Experiments presented in this figure were performed in the MOC1 model. A, B Higher panels; intensity of the different subpopulation clusters of CD8 (A) and CD4 (B) T cells identified using TriMap and FlowSOM algorithms, lower panels; heatmap showing the expression of the indicated markers in the different populations identified by FlowSOM algorithm across all treatment conditions (concatenated from Control n = 7, ATRi/RT n = 8, ATRi/RT/ImmuT n = 8; from one experiment). C Dot plot and donut chart representing % of CD8 and CD4_conv T cells in the different cell cycle phases across all conditions (concatenated from Control n = 5, ATRi/RT n = 6, ATRi/RT/ImmuT n = 6; from one experiment). Across all conditions the following graphs show % (D) PD-1⁺, (E) ICOS⁺, (F) naïve (N; CD62L⁺ CD44^-), effector (EM; CD62L^- CD44⁺ CD103^-) and tissue-resident (TRM; CD62L^- CD44⁺ CD103⁺) memory, (G) Ki67/PRF⁺ EM, (H) NKG2D⁺ and (I) T-BET⁺ CD8 and CD4_conv T cells (Control n = 15, ATRi/RT n = 17, ATRi/RT n = 17; combined from two independent experiments). Results are shown as means ± SEM and n represents number of mouse/groups. Parametric statistics were only applied to normally distributed data. Numbers on graphs represent P values and were determined by Kruskal-Wallis test with Dunn’s multiple comparison test (E; CD8 T cells; F; CD8 T_N, CD4conv T_N, CD8 T_EM, CD4_conv T_TRM; G; CD8 T cells) or ordinary one-way ANOVA with Tukey’s multiple comparison test (E; CD4_conv T cells, F; CD4_conv T_EM, CD8 T_TRM; G; CD4_conv T cells; H, I).