Fig. 2

Intratumoral administration of HER2 CAR T cells effectively treats medulloblastoma in an orthotopic model. a Outline of experimental setup. DAOY human medulloblastoma cells were transduced with lentivirus containing a GFP-Luciferase construct to facilitate in vivo study. Example DAOY-GL GFP expression is shown by flow cytometry. For the experiments, NOD.Cg-Prkdcscid Il2rgtm1Wjl/SzJ (NSG) mice were given tumor and treated with human CAR T cells intratumorally (IT) as shown. Bioluminescent imaging was performed using a Xenogen IVIS apparatus twice per week. b Tumor growth curves for NSG mice treated with either HER2 and CD19 CAR T cells intratumorally on Day 0. Results are shown for two experiments using T cells from two separate donors. Indicated doses correspond to the number of human CAR+ T cells infused into each animal. Error bars represent standard error of the mean. For shown statistics, data were normalized, linear regressions fitted to the normalized data, and slopes were compared using an ANOVA test. c Bioluminescent images from selected timepoints during the experiments. Indicated doses represent number of CAR positive T cells injected per mouse. d Brain and peripheral blood were harvested from mice on day 22 post- treatment and analyzed by flow cytometry. The presence of human CAR+ T cells was assessed by CD45 and Protein-L staining. Total amounts of human CAR T cells in the brain and relative concentrations in the peripheral blood for each group were compared using Mann-Whitney non-parametric statistical tests