Gene expression profile of canine Chimeric Antigen Receptor (CAR) Engineered invariant Natural Killer T (iNKT) cells

We used Nanostring gene expression profiling to characterize CAR-engineered canine iNKT cells The anti-IL13Rα2-28-ζ CAR construct was expressed along with a truncated canine CD19 marker. End-of-expansion, resting CAR-iNKTs were stimulated for 4-8 hours with anti-CD3/CD28-coated beads at 1:1 ratio. RNA was extracted using the RNeasy Plus Mini Kit (Qiagen) according to the manufacturer’s instructions. Gene expression profiling was performed using NanoString Canine Immuno-Oncology panel and nCounter instrument (Wistar Institute) according to the manufacturer’s instructions.