Fig 2: Tumor-specific CD8 T cell persist throughout tumorigenesis.A. Experimental scheme: naive TCRTAG (Thy1.1) were adoptively transferred into LMTAG-infected C57BL/6 (B6; Thy1.2) or tamoxifen-treated AST;Cre-ERT2 (Thy1.2) mice. TCRTAG were re-isolated from spleens and livers of infected mice (green) and tumor mice (blue) for flow cytometric analysis between 2.5–70+ days post-transfer. B. TCRTAG cell numbers in the spleens (left) and livers (right) of LMTAG-infected mice and tumor-bearing mice. Dots represent mean and error bars represent SEM. For B-D and F n = 2–3 mice per group (infection); n = 3 mice per group (tumor). Data is representative of two independent experiments. C. Histograms (left) showing TCRTAG PD1 expression from infection shown in comparison to naive (N; gray). For C-D and F, each symbol represents an individual mouse, and one-sample Student’s t-test with Bonferroni correction was used to determine significance: *, P<0.008 (infection); *, P<0.007 (tumor); ns, not statistically significant. D. Histograms and summary plots of CD38 expression and CD101 expression in spleens and livers from tumor mice shown in comparison to N. E. TCRTAG IFNγ and TNFα production after 4-hour ex vivo TAG peptide stimulation, with inset numbers indicating the percentage of cells in each gate. Gates were set based on no peptide stimulation controls. Right, summary plots of % IFNγ+ TCRTAG after peptide stimulation; each symbol represents an individual mouse; n = 3–4 mice per group. *, P<0.05; **, P<0.01; ****, P<0.0001 (one-way ANOVA with post hoc Tukey test). F. Histograms and summary plots of CD122 expression and BCL2 expression from tumor mice shown in comparison to N. All flow plots are gated on live CD8+ Thy1.1+ cells, and flow data for each timepoint is concatenated from all biological replicates. Summary plots (right) show MFI (geometric mean fluorescence intensity).