Extended Data Fig. 2. Orthogonal T-cell engineering enables ACT efficacy without lymphodepletion.

a Top: a representative first independent experiment comprising non-treated (n = 10) or treated mice with UT OT1 (n = 10), PD1d (n = 5 mice), IL-2^V(n = 4 mice/group), PD1d/2^V (n = 5 mice/group), PD1d/33 (n = 4 mice/group) and PD1d/2^V/33 (n = 6 mice/group). Bottom: a representative second independent experiment comprising non-treated (n = 6) or treated mice with UT OT1 (n = 8), UT OT1 plus αPD-L1 (n = 5 mice), PD1d/2^V (n = 9 mice/group), PD1d/33 and IL-33 (n = 5 mice/group), PD1d/2^V/33 (n = 8 mice/group). Data are presented as mean values ± s.d. Survival Data were determined using a log-rank Mantel−Cox test. Tumor growth comparison was done using a two-sided Mann–Whitney Test ^***P < 0.001, ^****P < 0.0001. b A representative experiment showing tumor growth curves of B16.OVA-bearing mice treated with either UT OT1 (n = 5 mice), UT OT1 plus αPD-L1 (n = 5) or PD1d/2v/33 (n = 10) on days 6 and 9 after tumor cell inoculation. c Survival curves of B16.OVA-bearing mice treated with either early or late orthogonal ACT from a–b. Data are presented as mean values ± s.d. Survival Data were determined using a log-rank Mantel−Cox test. Tumor growth comparison was done using a two-sided Mann–Whitney Test ^****P < 0.0001. d A representative experiment showing tumor growth and survival curves of non-treated MC38.OVA-bearing mice (n = 10) or treated with either UT OT1 (n = 10) or PD1d/2^V/33⁺OT1 cells (n = 10) on days 12 and 15 after tumor cell inoculation. e A representative experiment showing tumor growth and survival curves of B16.F10-bearing mice treated with either UT Pmel (n = 10) or PD1d/2^V/33⁺Pmel cells (n = 15) on days 12 and 15 after tumor cell inoculation Survival Data were determined using a log-rank Mantel−Cox test. Tumor growth comparison was done using a two-sided Mann–Whitney Test ^*P < 0.05, ^**P < 0.01, ^***P < 0.001, ^****P < 0.0001.

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