Protocol for assessing pharmacokinetics and pharmacodynamics of human CAR-NKT cells in humanized mouse models using bioluminescence imaging.

Human invariant natural killer T (NKT) cells exhibit strong tumor-killing ability and bridge innate and adaptive immunity. Peripheral blood mononuclear cell (PBMC)-derived chimeric antigen receptor (CAR)-engineered NKT (CAR-NKT) cells show potent anti-tumor activity. Here, we present a protocol for assessing the pharmacokinetics and pharmacodynamics (PK/PD) of PBMC-derived CAR-NKT cells in humanized mouse models using in vivo bioluminescence imaging (BLI). We describe steps for evaluating CAR-NKT cell distribution, persistence, and tumor infiltration across tumor-free and tumor-bearing models to support CAR-NKT cell therapy development. For complete details on the use and execution of this protocol, please refer to Li et al.(1)(,)(2).