Figure 1.

T cells with IL12-CAR executed their cytolytic functions in an antigen-dependent fashion

(A–C) (A) Schematic representation of the IL12-CAR. scFv, binding domain; hi, human IgG1 hinge; IL12, (p40-p35) IL12; IgG1-Fc, IgG1 CH2-CH3; CD28, transmembrane and intracellular domains without the lck binding motif; CD3ζ, intracellular domain of CD3ζ. The IL12 domain is of murine origin and binds to the human IL12 receptor; all other CAR domains are of human origin. IL12-CAR T cells with specificity for CEA (B) and Muc1 (C) or non-modified T cells (w/o) for control (0.31–5 × 10⁴ cells/well) were co-cultivated for 48 h in 96 micro-test plates with CEA⁺ Muc1^- LS174T, CEA⁻ Colo320 and Muc1⁺ MCF7 tumor cells (each 2.5 × 10⁴ cells/well), respectively. Viability of tumor cells was determined by the XTT assay and specific cytotoxicity was calculated. Numbers represent the mean values of technical replicates ±SD. Data from a typical experiment out of three are shown.