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Imaging Chimeric Antigen Receptor (CAR) Activation

  • Kendra A. Libby
  • Xiaolei SuEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 2111)

Abstract

The chimeric antigen receptor (CAR) has been extensively exploited in cancer immunotherapy. In spite of the success of CAR T cells in clinical applications, the molecular mechanism underlying CAR-T cell activation remains unclear. Key questions remain: how are CARs activated by tumor antigens? How do activated CARs transduce signaling to downstream pathways? Here we introduce a microscopy-based method for studying CAR signaling. We use an antigen-coated supported lipid bilayer to activate CARs and combine it with TIRF microscopy to visualize the initial activation process of CAR T cells. This enables monitoring CAR signaling at high spatial and temporal resolutions.

Key words

Chimeric antigen receptor T cell signaling Microclusters TIRF CD19 

Notes

Acknowledgments

This work was supported by a Yale College First-Year Summer Research Fellowship in the Sciences and Engineering awarded to Kendra Libby. Xiaolei Su was supported by Grant#IRG 17-172-57 from the American Cancer Society, Hood Foundation Child Health Research Award, Gilead Sciences Research Scholars Award in Hematology/Oncology, The Andrew McDonough B+ Foundation, and The Rally Foundation.

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2020

Authors and Affiliations

  1. 1.Department of Cell BiologyYale School of MedicineNew HavenUSA

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