Extended Data Fig. 4: Characterization of intratumoral CD8+ cell populations in AT3 tumors using HobitTomCreId3GFP double reporter mice.
From: Lymph-node-derived stem-like but not tumor-tissue-resident CD8+ T cells fuel anticancer immunity
(a-d) HobitTomCreId3GFP reporter mice were inoculated with AT3-OVA tumors for analysis of CD8+ T cells in tumors. Histograms and quantification of CD69 (a); CD39 and TOX (b), Ly108 and CX3CR1 (c) in ID3+ Hobit− (ID3 SP), Hobit+ ID3+ (DP), Hobit+ ID3− (Hobit SP), Hobit− ID3− (DN) from AT3-OVA tumors, pre-gated on CD44+ (n = 7) or tetramer+ (n = 9) cells. (d) HobitTomCreId3GFP reporter mice were inoculated with AT3-OVA tumors for analysis of CD8+ T cells in tumors. Histograms and quantifications showing the expression of IFNγ (n = 5), TNF (n = 6) and GzmB (n = 6) within ID3 SP, DP, Hobit SP and DN CD44+ subsets after PMA and ionomycin restimulation. (e-h) AT3-OVA tumor-bearing HobitTomCreId3GFP mice were treated with anti-PDL-1 and anti-CTLA-4 (ICB) every 2–3 days and CD8+ T cells in tumors analyzed 2–3 days after last treatment. (e) Tumor volume in untreated (n = 8) or ICB-treated (n = 8) mice over time. Flow cytometry plots (left), frequencies (middle), and numbers (right) of tumor of PD-1+ (untreated n = 13, ICB-treated n = 12) (f) and GzmB+ PD-1+ (untreated n = 6, ICB-treated n = 8) (g) TCF1+ PD-1+ (untreated n = 9, ICB-treated n = 7) (h) CD8+ T cells in tumors in untreated (n = 9) or ICB-treated (n = 7) mice. Flow cytometry plots are representative. Dots in graphs represent individual mice; horizontal lines and error bars of bar graphs indicate means and ±SEM. Data are pooled from at least two independent experiments. P values are from one-way ANOVA with Tukey’s comparisons test (a-d) or from two-tailed unpaired t-tests (e-h). P > 0.05, not significant (n.s.).
