Fig. 2
B7-H3-specific CAR T cells recognize and eradicate GBM cells. A Retroviral vectors construct encoding the B7-H3.CAR and bicistronic mCherry reporter. B Schematics of transduction and expansion of CAR T cells used in this study. C Expression of the B7-H3.CAR in transduced human T cells. The transduction efficiency of CAR molecules was measured either by mCherry expression or by direct staining of B7-H3 specific scFv molecules. Data shown are a representative FACS plot and quantifications of 3 independent donors. Error bars denote SEM. D Phenotypic analysis of CAR+ T cells at 12 days post transduction showing the frequency of CD4+ and CD8+ population in CAR+ T cells (left) as well as percentage of stem cell memory T cells (TSCM, CD45RA+CCR7+), central memory T cells (TCM, CD45RA−CCR7+), effector memory T cells (TEM, CD45RA−CCR7− ), and effector T cells (TE, CD45RA+CCR7− ) in CD4+ T cells and CD8+ T cells (n = 3). Error bars denote SEM. E Surface staining for CD69, CD25, and CD137 of CAR T cells after co-culture with the indicated cell lines for 24 h (n = 3). Error bars denote SEM. F Expression of granzyme B and cytokine (TNF-α, IFN-γ and IL-2) secretion of CAR T cells after co-culture with the indicated cell lines for 24 h (n = 3). Error bars denote SEM. G Counts of residual tumor cell lines after 5-day co-culture with CAR T cells or Ctrl T cells at 1:1 E: T ratio (n = 3). Error bars denote SEM. H&I Counts of U87 MG tumor cells and T cells (CAR T or Ctrl T cells) on indicated days post co-culture at 1:1 E: T ratio (n = 3). Error bars denote SEM. J Counts of U87 MG tumor cells on day two post co-culture with CAR T cells or Ctrl T cells at 1:1, 1:2, and 1:4 E: T ratio (n = 3). Error bars denote SEM