Abstract

The efficacy of chimeric antigen receptor (CAR) T?cell immunotherapy is limited by insufficient infiltration and activation of T?cells due to the immunosuppressive tumor microenvironment. Preclinical studies with optimized mouse CAR T?cells in immunocompetent mouse cancer models will help define the mechanisms underlying immunotherapy resistance. Here, we present a protocol for preparing mouse T?cells and generating CAR T?cells. We then detail procedures for testing their therapeutic efficacy and tracking them in a syngeneic mouse glioma model. For complete details on the use and execution of this protocol, please refer to Zhang et?al.¹.