Figure 3

CAR T cell-mediated killing of Nalm-6 cells in zebrafish. Zebrafish embryos were injected with Nalm-6 cells (green) at approximately 48 hpf. Around 2 hours later, either mock T cells (without a CAR) (red cells in (A)) or CD19 CAR T cells (red cells in (B)) were injected and images were recorded within 2 hours and again at 24 hours post injection of T cells. (C) The numbers of Nalm-6 cells and T cells were quantified at 2 hpi and 24 hpi based on the fluorescent area covered by cells in the tail (red box in B) using Fiji. The two time points are connected by a line for each embryo. From left to right: Nalm-6 cells in zebrafish without any T cells; Nalm-6 cells (in zebrafish with CD19 CAR T cells), CD19 CAR T cells (in zebrafish with Nalm-6); Nalm-6 cells (in zebrafish with mock T cells), mock T cells (in zebrafish with Nalm-6) at 2 hpi and 24 hpi. (D) Violin plots normalized to the area covered by fluorescent cells at 2 hpi revealing the change in distribution at 24 hpi. Nalm-6 cells together with mock T cells or alone without T cells show similar distributions at 24 hpi, whereas Nalm-6 injected with CD19 CAR T cells show a reduction compared to Nalm-6 alone or Nalm-6 with mock T cells. Images were recorded on a Zeiss Axio Zoom.V16 fluorescence stereo zoom microscope and rendered with Adobe Photoshop CS6. Scale bar in (A) represents 1 mm.