1. Fig. 1. Selection of a 4-1BB-endodomain–based anti-EGFRvIII CAR construct.
Selection of a 4-1BB-endodomain–based anti-EGFRvIII CAR construct.(A) Vector maps of tested anti-EGFRvIII CAR designs. (B) Flow-based cytotoxicity of CART-EGFRvIII cells against EGFRvIII+ glioma cell line. The glioblastoma cell line U87MG was stably transduced with EGFRvIII, fluorescently labeled with CFSE, and cocultured for 18 hours. Untransduced (UTD) or CD19.BBz CAR T cells were used as negative controls and compared to 3C10.BBz, 3C10.28BBz, or 139.BBz EGFRvIII CAR T cells. Lysis of parental cells (U87MG) and EGFRvIII-transduced U87 cells was analyzed at different effector/target (E:T) ratios by flow cytometry. One representative experiment is shown. Samples were performed in duplicate in two replicative experiments. (C) CART-EGFRvIII cells eradicate glioblastoma in an orthotopic xenogeneic mouse model. Seven days after 5 × 104 U87-EGFRvIII cells were orthotopically implanted into mouse brains, mice were injected intravenously with either phosphate-buffered saline (PBS) alone, temozolomide (TMZ) alone, or temozolomide with 1 × 106 transduced T cells expressing the indicated CAR constructs. Tumor burden was quantified as total flux in units of photons/second. Bars indicate means ± SD (n = 10 mice per group). Note that all mice in the PBS group died by day 15.
Laura A. Johnson et al., Sci Transl Med 2015;7:275ra22
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