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. 2023 Jan 4;120(2):e2216352120. doi: 10.1073/pnas.2216352120

Fig. 1.

Fig. 1.

CARs show reduced sensitivity compared to the TCR when antigen is presented on APCs but not when presented as purified protein. (A) Schematic of antigen receptors. The 1G4 TCR and the D52N scFv both recognize the 9V NY-ESO-1 peptide antigen presented on HLA-A*02:01. CARs using the CD8a hinge contain the CD8a transmembrane domain, whereas CARs using the IgG1 or CD28 hinges contain the CD28 transmembrane domain. (B) Schematic of APC stimulation system. (C and D) Representative dose–response showing (C) cytotoxicity by LDH release and (D) surface expression of CD69 for the TCR and the indicated CARs along with EC50 values from at least three independent experiments determined by fitting a Hill function to each dose–response curve. (E) Representative dose–response when the purified biotinylated 9V pMHC ligand is presented on streptavidin-coated plates (Left two plots) and EC50 values from at least three independent experiments (Right). The EC50 values are compared using (C and D) one-way ANOVA or (E) one-sample t-test for a hypothetical mean of 1.0 on log-transformed values. *P-value ≤ 0.05, **P-value ≤ 0.01, ***P-value ≤ 0.001, ****P-value ≤ 0.0001.