Figure 3.

Control of CAR T-cell function by CRASH-IT. A, Schematic diagram of second-generation anti-CD19-CD28-CD3ζ CAR interacting with the Zap70-PD1-SMASh construct. B, CD19 expression on K562, Daudi, and Raji tumors. Cells were stained with anti-CD19-PE (red line) or isotype control-PE (black line). C–F, Primary human T cells modified with the anti-CD19-CD28-CD3ζ CAR and either Zap70-PD1-SMASh (C and E), or vector control (D and F) were pretreated with 10 μmol/L asunaprevir (ASUNA) or DMSO control. Data depict intracellular IFNγ, IL2, TNFα and cell-surface LAMP1 expression of CAR⁺, EGFP-high, CD8⁺ (C and D) and CD4⁺ (E and F) T cells upon coculture with CD19⁻ K562 tumor cells, or CD19⁺ Daudi or Raji tumor cells in the continued presence or absence of asunaprevir. Error bars represent SD (n = 3). Data are representative of two independent experiments.