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Analysis of CAR-Mediated Tonic Signaling

  • Hugo Calderon
  • Maksim Mamonkin
  • Sonia GuedanEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 2086)

Abstract

CARs are synthetic receptors designed to drive antigen-specific activation upon binding of the scFv to its cognate antigen. However, CARs can also elicit different levels of ligand-independent constitutive signaling, also known as tonic signaling. Chronic T cell activation is observed in certain combinations of scFv, hinge, and costimulatory domains and may be increased due to high levels of CAR expression. Tonic signaling can be identified during primary T cell expansion due to differences in the phenotype and growth of CAR-T cells compared to control T cells. CARs displaying tonic signaling are associated with accelerated T cell differentiation and exhaustion and impaired antitumor effects. Selecting CARs which configuration does not induce tonic signaling is important to enhance antigen-specific T cell responses. In this chapter, we describe in detail different protocols to identify tonic signaling driven by CARs during primary T cell ex vivo expansion.

Key words

Chimeric antigen receptors (CAR) Adoptive T cell transfer (ACT) Tonic signaling T cell exhaustion Differentiation Antigen-independent signaling Chronic activation 

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Copyright information

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

Authors and Affiliations

  1. 1.Department of Hematology and Oncology, Hospital ClinicInstitut d’Investigacions Biomèdiques August Pi i Sunyer (IDIBAPS)BarcelonaSpain
  2. 2.Center for Cell and Gene TherapyBaylor College of MedicineHoustonUSA

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