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Imaging CAR-T Synapse as a Quality Control for CAR Engineering

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The Immune Synapse

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2654))

Abstract

The chimeric antigen receptor (CAR) evolves as a powerful tool to reprogram T cells for targeted killing. CAR-T therapy succeeded in treating certain types of blood cancers, and its application is now expanding towards solid tumors, autoimmune diseases, viral infection, and fibrosis. These require the design of a large number of new CARs that target a variety of antigens. Here we described two methods as a quality control for validating newly developed CARs: (1) the cell-cell conjugation assay as a reflection of efficient binding of CAR to antigen in the cellular context and (2) CD45 exclusion in the synapse as an indication of CAR signaling potential. These assays examine prerequisites for a functional CAR-T and reveal causes for ineffective CAR-T activation.

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Author information

Authors and Affiliations

  1. Department of Cell Biology, Yale School of Medicine, New Haven, CT, USA

    Qian Xiao & Xiaolei Su

  2. Duncan and Nancy MacMillan Cancer Immunology and Metabolism Center of Excellence, Rutgers Cancer Institute of New Jersey, New Brunswick, NJ, USA

    Qian Xiao

  3. Department of Medicine, Robert Wood Johnson Medical School, Rutgers University, New Brunswick, NJ, USA

    Qian Xiao

  4. Yale Cancer Center, Yale University, New Haven, CT, USA

    Xiaolei Su

Authors
  1. Qian Xiao
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  2. Xiaolei Su
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Corresponding author

Correspondence to Xiaolei Su .

Editor information

Editors and Affiliations

  1. Department of Life Sciences, University of Siena, Siena, Italy

    Cosima T. Baldari

  2. Kennedy Institute of Rheumatology, University of Oxford, Oxford, UK

    Michael L. Dustin

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© 2023 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature

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Xiao, Q., Su, X. (2023). Imaging CAR-T Synapse as a Quality Control for CAR Engineering. In: Baldari, C.T., Dustin, M.L. (eds) The Immune Synapse. Methods in Molecular Biology, vol 2654. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3135-5_33

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  • DOI: https://doi.org/10.1007/978-1-0716-3135-5_33

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-3134-8

  • Online ISBN: 978-1-0716-3135-5

  • eBook Packages: Springer Protocols

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