Figure 3.

Proliferation, activation and cytotoxicity of EGFR CAR‐T cells. (a) Primary T lymphocytes infected with EGFR CAR lentivirus (EGFR CAR‐T) labelled with CFSE were incubated with or without MDA‐MB‐231 or MDA‐MB‐468 cells in culture medium without adding proliferative cytokines for 3 days and diluted to examine their proliferation. Experiments were repeated three times, and representative histograms are shown. (b, c) CTL T or EGFR CAR‐T cells were incubated with MDA‐MB‐231 or MDA‐MB‐468 cells and stained with CD8‐APC and CD69‐PE (b) or CD25‐FITC (c) followed by flow cytometry analysis. Experiments were repeated three times, and representative histograms are shown. (d) EGFR CAR‐T cells were incubated with MDA‐MB‐231 or MDA‐MB‐468 cells at the indicated ratios for 3 days before measuring the secretion of cytokines, including IL‐2, TNFα and IFNγ. CTL T cells were used as a negative control. Data were obtained from three replicates and are presented as mean ± s.e.m.. (e, f) CTL T or EGFR CAR‐T cells were incubated with MDA‐MB‐231 (e) or MDA‐MB‐468 (f) cells at different ratios for the indicated durations followed by the cytotoxicity assay. Data were obtained from three replicates and are presented as mean ± s.e.m.. (g) EGFR CAR‐T cells were incubated with MCF10A, MCF7, BT474, MDA‐MB‐468 or MDA‐MB‐231 cells at a ratio of 1:1 for the indicated durations followed by the cytotoxicity assay. Data were obtained from three replicates and are presented as mean ± s.e.m.. CAR‐T, chimeric antigen receptor‐modified T cells; CFSE, carboxyfluorescein succinimidyl amino ester; CTL T, control T cells; EGFR, epidermal growth factor receptor; IFNγ, interferon γ; IL‐2, interleukin 2; TNFα, tumor necrosis factor α.