Characterization of CD8⁺ BM T cells in AML at the protein level and importance of Early Tm/Term imbalance. (A) Heatmap with the fluorescence intensity values of 20 markers across 5 CD8⁺ BM subsets: naïve (CCR7, CD27, CD28, CD127, and TCF1), Early Tm (CD27, CD28, TCF1, GZMK, CD127, PD-1, and TIGIT), Act (CD38, CD69, GZMK, PD1, and TIGIT), Int (GZMB and Tbet), and Term/SenL (CD57, KLRG1, CX3CR1, and GZMB). Median marker expression identifies the markers that characterize each CD8⁺ T-cell subset. (B) UMAP plots color-coded by selected single markers’ expression. (C) UMAP plots color-coded by subsets defined in panel A, split by groups and time points. (D) PCA depicting 2 predicted developmental trajectories via Slingshot, colored by CD8⁺ subsets for the first 3 principal components. In each pairwise combination (PCA_1-PCA_2, PCA_1-PCA_3, and PCA_2-PCA_3), the CD8⁺ T-cell differentiation path originates from the naïve subset and diverges after the Early Tm subset. The trajectories lead to 2 terminal end states: Act and Term/SenL. (E) Box plot showing the distribution of the CD8⁺ T-cell subsets along the inferred trajectories. (F) Contour plots illustrating Early Tm subset gating and percentage among memory CD8⁺ T cells. (G) Box plot showing proportion of the Early Tm subset in Res and NonRes at bas. Statistical significance was examined using a 2-sample Student t test. (H) Kaplan-Meier estimates of OS by high vs low Early Tm/Term ratio calculated using the median split. Statistical significance was calculated using the log-rank test. PCA, principal component analysis.