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

用于活细胞成像以观察免疫突触形成和CAR T细胞抗癌细胞激活的方案

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作者:Jorge Ibanez ,Nikhil Hebbar ,Unmesha Thanekar ,Zhongzhen Yi ,Haley Houke ,Meghan Ward ,Chris Nevitt ,Liqing Tian ,Stephen C Mack ,Heather Sheppard ,Jason Chiang ,M Paulina Velasquez ,Giedre Krenciute

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. Keywords: cancer; immunology; microscopy.

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