Figure 1.

Differential cellular composition and lineage of AB680 treated, and programmed cell death protein 1 (PD-1) blocker treated and untreated tumor-infiltrating lymphocytes (TILs). (A) Workflow of the study design. Single-cell RNA sequencing (scRNA-seq) was conducted with CD45⁺ TIL cells extracted from three untreated controls, AB680-treated and PD-1 blocking antibody-treated tumors. Eight tumors per group were used for fluorescence-activated cell sorting (FACS) analysis. (B) Five days after inoculation of CT26 cells (1×10⁶ cells/mouse), AB680 (20 mg/kg) or the PD-1 blocking antibody (20 mg/kg) was administered from day 0 to day 15 after initiation of treatment via the intraperitoneal route, as described in the Materials and methods section (n=9 mice). Tumor sizes were measured at day 3. (C) At day 15, mice were sacrificed and the extracted tumors were weighted. (D) Uniform manifold approximation and projection (UMAP) plot for clusters including all samples. (E) Proportion of clusters in each sample. (F) Heatmap representing cluster-specific gene expressions. The yellow color represents high expression, and purple represents low expression. (G) Proportions of clusters among the controls, AB680-treated, and PD-1 blocker treated TILs. The error bar denotes SEM (t-test, *p<0.05, **p<0.05, ***p=0.001). NS, non-significant.