Extended Data Fig. 9: Dual inhibition of BTLA and PD-1 in CAR T cells.

a,b, Data from Fig. 2j,m now including PD1 KO condition (n = 8 mice). c. Data from Fig. 3i, now including PD1 KO condition (n = 7 mice). d-k, Dual checkpoint inhibition of BTLA and PD-1 in CAR30 T cells promotes potent tumor control. d. DepMap Expression Score of HVEM and PDL1 in select CD30⁺ cell lines. e, 72 h killing of UTD or CAR30 T cells with Pembrolizumab (Pembro) against HDLM-2 (effector: target = 0.25, n = 3 technical replicates per condition). f, Proposed schema for dual BTLA/PD-1 inhibition in CAR30 T cells using CRISPR-Cas9-mediated deletion of BTLA and blockade of PD-1 using Pembro g, Design of in vivo study using HDLM-2 and NSG mice (Methods). Experimental arms were UTD (n = 4), UTD + Pembro (n = 4), WT CART30 + Saline (n = 4), WT CART30 + Pembro (n = 6), BTLA KO CART30 + Saline (n = 5), or BTLA KO CART30 + Pembro (n = 6). h, Median bioluminescent intensity (tumor burden) of each condition, with individual mouse curves shown in the background. i, Overall survival. j, Peripheral blood human CD45⁺ CD3⁺ counts on day 16 post-CART30 infusion. k, Tumor killing at 48 h of coculture, using WT or BTLA KO CAR30 T cells against KM-H2 (top) or SU-DH-L1 (bottom), in the prescence of either Vehicle (saline) or Pembro (100 μg / mL (effector: target ratio = 0.125, n = 3 technical replicates). l, Expression of BTLA and PD-1 in CD4⁺ and CD8⁺ pan-cancer TILs from the online portal of Zheng and colleagues (http://cancer-pku.cn:3838/PanC_T/)⁴¹. For all bar plots, data are presented as mean ± s.d. Statistical significance was determined by one-way ANOVA with post hoc Tukey tests (a (week 9 only),e,j,k), or long-rank Mantel Cox test (c,i).

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