CD1a CARTs derived from patients with coT-ALL at presentation (specifically lyse autologous CD1a⁺ T-ALL blasts). (A) Scheme depicting the experimental design for the autologous cytotoxic assay. Mature (normal) CD3⁺CD1a^– T cells were FACS-purified from the PB of a patient with coT-ALL, infected with CD1a CAR, expanded, and exposed to autologous total PBMCs. (B) FACS analysis of autologous cytotoxic 48-hour assay at 1:1 and 4:1 E:T ratios. eFluor 670–labeled total PBMC target population contains CD1a⁺ T-ALL blasts (red) and mature CD3⁺CD1a^– T cells (blue). (C) Quantification of CD1a CART-mediated specific lysis for coT-ALL blasts (upper panel) and CD3⁺CD1a^– mature T cells (bottom panel). (D) ELISpot showing the number of IFN-γ SFC from mock vs CD1a CARTs stimulated with a pool of peptides from CMV, EBV, and flu (CEF). Staphylococcal enterotoxin B was used as positive control. SFC, spot-forming cell.