Enhanced tumor protection in a colorectal model through adoptive transfer of CD3⁺ T cells from mice treated with RT plus CART-19. (A) Working model: time line and schematic representation of in vivo A20 tumor–bearing mice treated with 2 × 4-Gy RT followed by CAR T cells and adoptive T-cell transfer to CT26 tumor–bearing mice. All mice of all groups were treated with the same CP lymphodepletion regimen. (B) AH1-specific T cells from donor mice after ex vivo expansion, before ACT. (C) CT26 tumor growth of recipient mice (n > 4). (D) CD45⁺/CD3⁺/CD8⁺ T-cell tumor infiltration. (E) CD45⁺/CD3⁺/CD8⁺/CD107⁺ T-cell tumor infiltration. (F) CD45⁺/CD3⁺/CD8⁺/AH1-specific T-cell tumor infiltration. (G) IFN-γ spots of an ELISPOT assay after AH1 peptide stimulation of splenocytes from mice that received ACT. Graphs show the mean ± SEM. ∗P < .05; ∗∗P < .01; ∗∗∗P < .001; ∗∗∗∗P < .0001. Data are representative of 2 independent experiments. ACT, Adoptive Cell Therapy.