Comparative analysis of assays to measure CAR T-cell-mediated cytotoxicity Review

   


Authors: Kiesgen, S.; Messinger, J. C.; Chintala, N. K.; Tano, Z.; Adusumilli, P. S.
Review Title: Comparative analysis of assays to measure CAR T-cell-mediated cytotoxicity
Abstract: The antitumor efficacy of genetically engineered ‘living drugs’, including chimeric antigen receptor and T-cell receptor T cells, is influenced by their activation, proliferation, inhibition, and exhaustion. A sensitive and reproducible cytotoxicity assay that collectively reflects these functions is an essential requirement for translation of these cellular therapeutic agents. Here, we compare various in vitro cytotoxicity assays (including chromium release, bioluminescence, impedance, and flow cytometry) with respect to their experimental setup, appropriate uses, advantages, and disadvantages, and measures to overcome their limitations. We also highlight the US Food and Drug Administration (FDA) directives for a potency assay for release of clinical cell therapy products. In addition, we discuss advanced assays of repeated antigen exposure and simultaneous testing of combinations of immune effector cells, immunomodulatory antibodies, and targets with variable antigen expression. This review article should help to equip investigators with the necessary knowledge to select appropriate cytotoxicity assays to test the efficacy of immunotherapeutic agents alone or in combination. © 2020, Springer Nature Limited.
Keywords: flow cytometry; t lymphocyte; t-lymphocytes; animal; metabolism; animals; drug screening; xenograft model antitumor assays; immunology; receptors, antigen, t-cell; adoptive immunotherapy; immunotherapy, adoptive; lymphocyte antigen receptor; cytotoxicity tests, immunologic; cytotoxicity test; procedures; humans; human; receptors, chimeric antigen
Journal Title: Nature Protocols
Volume: 16
Issue: 3
ISSN: 1754-2189
Publisher: Nature Publishing Group  
Date Published: 2021-03-01
Start Page: 1331
End Page: 1342
Language: English
DOI: 10.1038/s41596-020-00467-0
PUBMED: 33589826
PROVIDER: scopus
PMCID: PMC8064272
DOI/URL:

https://www.scopus.com/inward/record.uri?eid=2-s2.0-85101446722&doi=10.1038%2fs41596-020-00467-0&partnerID=40&md5=72c1edd74936c26e378da519e9cbf3a6
Notes: Review -- Export Date: 1 June 2021 -- Source: Scopus
Altmetric
What is Altmetric?
Citation Impact
What is Dimensions Citation Badge?
BMJ Impact Analytics
MSK Authors
  1. Prasad S Adusumilli
    494 Adusumilli
  2. Navin Kumar Chintala
    27 Chintala
  3. Zachary Tano
    11 Tano
  4. Stefan Thomas Kiesgen
    5 Kiesgen
  5. John Conrad Messinger
    7 Messinger
Related MSK Work