Protocol for live-cell imaging of immune synapse formation and activation of CAR T cells against cancer cells.

Abstract

Immune synapse (IS) formation determines T cell antitumor activity. Here, we present a protocol for characterizing the IS formation between chimeric antigen receptor (CAR) T cells and tumor cells by measuring the IS size and calcium flux by live-cell imaging. We describe steps for CAR T cell manufacturing, sample preparation, image acquisition, and data analysis. For complete details on the use and execution of this protocol, please refer to Chockley et al.,¹ Ibanez et al.,² and Zoine et al.³.