Fig. 5. PD-1 decoy is dispensable for orthogonal ACT.

a, Tumor growth control over time of B16-OVA tumor-bearing mice treated with PD1d/IL-2v/IL-33⁺ OT1 cells in the presence or absence of 250 μg per mouse of antibodies specific for the indicated immune checkpoints administered intraperitoneally (i.p.) beginning 1 d before the first cell transfer and maintained every 3 d for a maximum of six doses. b, Tumor growth control over time of B16-OVA tumor-bearing mice treated with PD1d/IL-2v/IL-33⁺ OT1 cells or with OT1 T cells genetically engineered to secrete only IL-2v and IL-33 (no PD-1 ectodomain). c, Percentage of Gmzc⁺TCF1^neg effector-like cells in CD44⁺PD-1⁺ OT1 TILs recovered from mice treated with either PD1d/IL-2v/IL-33⁺ OT1 cells or IL-2v/IL-33 OT1 cells 12 d after orthogonal ACT (two independent experiments, n = 4 mice per group per experiment). Data are presented as mean values ± s.d. d, Expression of effector molecules in CD44⁺PD-1⁺Gzmc⁺TCF1^neg effector-like cells in OT1 TILs from b. A two-tailed Student’s t-test with Welch’s correction was used for comparing both groups in c and d. NS, P > 0.05. A representative experiment of two independent experiments (n = 6 animals per group) is shown for experiments in a and b. e, GSEA: CD8⁺ T-cell gene signatures of PD-1-mediated inhibition³⁷. Ranked list: genes differentially expressed between C5 T_SE and canonical T_EX from the TIL reference map²¹.

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