Fig. 6.

Activity of 237-derived CAR T cells against additional human Tn-peptides. (A) To assess activation, mock, 237-CAR, TNGK-CAR, or 5E5-CAR transduced T cells were added to plates containing various concentrations of the indicated Tn-peptides and incubated at 37 °C, 5% CO₂. After 24 h, IFN-γ released in the culture supernatants was measured by ELISA. Activation of transduced cells under each condition was measured in duplicate in two independent experiments. Error bars indicate SEM. (B) Sensitization doses that yielded half-maximal stimulation (SD₅₀) of IFN-γ release were calculated for the active Tn-peptides using nonlinear regression analysis (GraphPad Prism 5). 5E5-CAR was not activated with TFRC and ZIP6; hence the SD₅₀ values were assigned values of >10⁻⁴ M (indicated by an *). SD₅₀ values from two independent experiments are plotted as replicates. (C) Maximum IFN-γ release for the active peptides was determined at the 1-μM concentration. Mean ± SD is plotted for B and C.