Supplementary Figure 4: Quantification of tumor-infiltrating and circulating CAR T cells in mice with brain tumors.
From: CAR-T cells secreting BiTEs circumvent antigen escape without detectable toxicity
(a) H&E and (b) Human CD3 IHC of brain tumors (U87vIII) from mice treated with CART-EGFRvIII.BiTE-EGFR cells. (c) U87KO (EGFR-negative), U87 or U87vIII gliomas were implanted intracranially in NSG mice. Mice were infused intraventricularly on day 14 post-implantation with CART-EGFRvIII.BiTE-EGFR cells. Brain tumors were isolated on day 7 post-treatment and assessd for CAR-transduced cells (mCherry), (d) ddPCR for the 4-1BB-CD3ζ transgene and (e) human CD3+ events. (f) Circulating blood was also assessed by flow cytometry and (g) ddPCR. (h) Gating strategy for infiltrating and (i) circulating CAR T cells. The experiment was repeated with similar results. Representative data are shown, n = 3 biologically independent animals (mean + SEM is depicted; unpaired, one-tailed t-test, * = p < 0.05, ** = p < 0.01, *** = p < 0.001).
