OrexiCAR T cells overcome M2-mediated immunosuppression. (A) Schematic of experimental set up. (B) T cells, M2s, and Raji-GFP/Luc tumor cells were cocultured for 4 and 24 hours. Tumor lysis was measured using luminescence. (C) Same experiment in panel B, which compared effect of M2s on T-cell–mediatedtumor lysis. (D) M2s alone were cultured with Raji tumor cells at indicated ratios in either WT or OrexiCAR supernatant ± rituximab (1μg/mL). (E-F) Supernatants from the indicated experiments were harvested after 24 hours and analyzed using Luminex for IL-10 (E) and IFN-γ (F). (G) M2s alone were cultured with Raji tumor cells in WT supernatant, WT supernatant supplemented with recombinant CV1 in excess, or OrexiCAR supernatant for 4 hours, and tumor lysis was measured. (H) WT CAR T cells, M2s, and Raji tumor cells were cocultured for 4 hours in the indicated supernatant. Data shown are from 1 representative human donor; all experiments were corroborated with 2 or more donors. Statistics were performed using student t test. –mac, no macrophages present; mac, macrophage; NT, nontransduced T cells; ritux, rituximab; sup, supernatant harvested from transduced T cells.