Figure 4.PD-1 antibodies enhanced αROR1-CAR T killing of A549 lung cancer cells under elevated pressure. (A–B) A549-Red-Fluc cells preconditioned in elevated pressure (+100 mmHg) for 7 days were incubated with non-preconditioned αROR1-CAR T cells under normal conditions. PD-L1 blocking antibodies Pembrolizumab (Pembr) and Cemiplimab (Cemipl) were added at 10 μg/ml at the same time. Cytotoxicity at indicated time points was calculated based on luciferase activity. (C–D) Cytokines (IFN-γ and TNF-α) in the medium of co-culture shown in (A) and (B) were measured by ELISA. (E) Flow cytometry analysis of co-cultures in (A) and (B) at 4 hours. Annexin V indicates apoptosis and Propidium iodide indicates cell death. Shown are representative results of 3 biological repeats. (F–G) Quantification of apoptosis and cell death by 3 biological repeats of experiments in (E).
