Extended Data Figure 9. Co-delivery of ORFs with CD19-targeting CARs.

a) Transduction efficiency of CAR+ORF lentiviral vectors or ORF alone (n = 4 biological replicates). b,c) CAR expression level as determined by staining with anti-mouse Fab F(ab’)₂-Representative histograms (b) and quantification of CAR expression relative to tNGFR (c) is shown for two healthy donors and two diffuse large B-cell lymphoma (DLBCL) patients, d) Expansion curves of CAR+ORF transduced T-cells (n = 4 biological replicates). e) LTBR expression in autologous CD14+ monocytes and T-cells transduced with LTBR alone or CAR+LTBR. f-i) Expression of ICAM-1 (f), CD70 (g), CD74 (h) and MHC-II (i) by T-cells transduced with LTBR ORF only, CAR+LTBR or CAR+tNGFR. All data are normalised to tNGFR only (no CAR). Unpaired two-sided t test p values are shown. j-m) Expression of exhaustion markers PD-1 (j), TIM-3 (k), LAG-3 (l) and CD39 (m) in CAR+ORF T-cells. n) Differentiation phenotype of CAR+ORF T-cells. CM: Central memory. EM: Effector memory. Differentiation was defined based on CD45RO and CCR7 expression (naïve: CD45RO^neg CCR7⁺, CM: CD45RO⁺ CCR7⁺, EM: CD45RO⁺ CCR7^neg, effector CD45RO^neg CCR7^neg). o,p) Expression of activation markers CD25 (o) and CD69 (p) in CAR+ORF T-cells incubated alone or with Nalm6 cells for 24 hours.

Error bars indicate SEM. N = 3 biological replicates, unless indicated otherwise.