Memory and effector T-cell activation, TCR signaling, and IFN-γ/IL-12 signaling pathways were implicated in T-cell immunity against ALL relapse in mice. (A) The schema shows the design of molecular profiling experiments. Five-week dasatinib therapy was administered to B6 mice 2 weeks after leukemia cell inoculation. Mice that remained in remission after 100 days were considered cured (ie, the cured group). By contrast, mice that experienced relapse were labeled the relapsed group. Specimens from each group were subjected to 3 assays: (1) serum was collected after 1 week of dasatinib treatment and used for multiplex assay to quantify 23 cytokines (cured, n = 3; relapsed, n = 7); (2) peripheral blood was collected after 1 week of dasatinib treatment, CD4 or CD8 T cells were isolated using flow cytometry, and RNA was purified and used for microarray gene expression profiling (cured, n = 4; relapsed, n = 4); and (3) at this same time point, peripheral blood leukocytes were collected and subjected to single-cell RNA sequencing (scRNA-seq; cured, n = 6; relapsed, n = 6). (B) Volcano plot shows differentially expressed genes in CD8 T cells (left panel) or CD4 (right panel) T cells from mice in the cured vs relapsed group. Green indicates genes upregulated in the cured group, and brown highlights genes upregulated in relapsed mice. Gray dash lines indicate P = .05 (y-axis) and log₂ (fold change) = ±0.5 (x-axis). (C) Heat map shows the gene sets enriched in T cells collected from the cured or relapsed group of mice. Analysis was performed for CD8 or CD4 T cells separately. Color indicates the degree of enrichment (normalized enrichment score), and blank means nominal P > .05.