Figure 4. Variations in ROR1-CAR targeting domain affect antitumor function in 3D lung cancer models.

(A) Quantification of apoptosis induced by ROR1-CAR T cells with either 2A2 or R12 targeting domains during 72-hour treatment with increasing CD8⁺ T cell dose (5 × 10⁴, 2.5 × 10⁵, 1 × 10⁶). Apoptosis was measured with M30 ELISA from supernatants collected at the indicated time points and is presented as fold change compared with the same dose of control T cells (red line). Data are presented as arithmetic mean of 3 cell crowns ± SD. n = 1 experiment. (B) ELISA-based quantification of IFN-γ and IL-2 from supernatants collected at the indicated time points from static tumor models treated with increasing CD8⁺ T cell numbers for 72 hours. Data are presented as arithmetic mean of 3 cell crowns ± SD. n = 1 experiment. (C) Expression of CD25 and CD69 on CD8⁺ ROR1-CAR T cells and unmodified control T cells at the end of the 72-hour analysis period in the static tumor model. One representative plot of 3 cell crowns from n = 1 experiment is shown.