Figure 3
From: Flexible Targeting of ErbB Dimers That Drive Tumorigenesis by Using Genetically Engineered T Cells
ErbB specificity of the T1E28z CAR. (A) ErbB null 32D myeloid cells were engineered to express all 10 ErbB receptor/dimer combinations. Expression of the indicated ErbB receptor was detected by FACS (filled histogram represents staining of parental 32D cells with the same antibody combination) and, in the case of ErbB3, by Western blotting (B). (C) To define targeting specificity, 1 × 106 of the indicated engineered T-cell populations was cocultivated with an equal number of 32D cells that express the specified ErbB receptor(s). Supernatants were harvested at 24 h (IL-2) and 72 h (IFN-γ) for ELISA analysis. Data shown are mean ± standard deviation (SD) of three (IL-2) or six (IFN-γ) replicates after correction for transduction efficiency between groups. ***P < 0.001, **P < 0.01, comparing T1E28z (black) or EGF28z (gray) with other control groups. (D) 32D cells that express ErbB1 (32D:1) or ErbB2 + 3 (32D:23) were cocultivated with T1E28z+ T cells in the presence of exogenous EGF (left) or HRG-1β (right). Supernatants harvested at 72 h were analyzed for IFN-γ content.