Fig. 3

Type I interferon and immune cell profiles in the tumor microenvironment. a. Spleen cells from tumor-free mice were stained with CD4- and CD8-specific mAbs and analyzed by flow cytometry. Shown are representative flow cytometry plots. The % CD4⁺ and CD8⁺ T cells were quantified and presented at the right panel. b. Cells were prepared from spleens of the MC38 tumor-bearing WT (n = 5) and IFNAR1-KO (n = 5) mice as shown in Fig. 2B. Top panel shows gating strategy for spleen cells. Single cells were gated out for live and dead cells. The live cells were gated for CD4⁺ and CD8⁺ T cells as in A. c. Gating strategy for tumor-infiltrating immune cells. Tumor were excised from the MC38 tumor-bearing WT (n = 5) and IFNAR1-KO (n = 5) mice, digested with collagenase to make single cells. The digested tumor cell mixtures were stained with Zombie violet and CD4-, CD8-, CD11b-, and Gr1-specific mAbs. The digested cells were gated for single cells, followed by gating for live and dead cells. d & e. The live cells as shown in C were analyzed for T cells (d) and CD11b⁺Gr1⁺ cells (e) by flow cytometry. Shown at the left panels are representative plots of one pair of mice. The % tumor-infiltrating CD4⁺ CD8⁺ T cells (d) and CD11b⁺Gr1⁺ cells (e) were quantified and presented at the right panel