High-dimensional functional phenotyping of preclinical human CAR T cells using mass cytometry

Ilaria M. Michelozzi; Jahangir Sufi; Thomas A. Adejumo; Persis J. Amrolia; Christopher J. Tape; Alice Giustacchini

STAR Protocols (Mar 2022)

High-dimensional functional phenotyping of preclinical human CAR T cells using mass cytometry

Ilaria M. Michelozzi,
Jahangir Sufi,
Thomas A. Adejumo,
Persis J. Amrolia,
Christopher J. Tape,
Alice Giustacchini

Affiliations

Ilaria M. Michelozzi: Molecular and Cellular Immunology Section, UCL Great Ormond Street Institute of Child Health, Zayed Centre For Research into Rare Disease in Children, WC1N 1DZ London, UK; Corresponding author
Jahangir Sufi: Cell Communication Lab, Department of Oncology, University College London Cancer Institute, WC1E 6DD London, UK
Thomas A. Adejumo: Fluidigm Corporation, South San Francisco, CA, USA
Persis J. Amrolia: Molecular and Cellular Immunology Section, UCL Great Ormond Street Institute of Child Health, Zayed Centre For Research into Rare Disease in Children, WC1N 1DZ London, UK; Department of Bone Marrow Transplant, Great Ormond Street Hospital for Children, WC1N 3JH London, UK
Christopher J. Tape: Cell Communication Lab, Department of Oncology, University College London Cancer Institute, WC1E 6DD London, UK
Alice Giustacchini: Molecular and Cellular Immunology Section, UCL Great Ormond Street Institute of Child Health, Zayed Centre For Research into Rare Disease in Children, WC1N 1DZ London, UK; Corresponding author

Journal volume & issue: Vol. 3, no. 1
p. 101174

Abstract

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Summary: Here, we present a comprehensive protocol for the generation and functional characterization of chimeric antigen receptor (CAR) T cells and their products by mass cytometry in a reproducible and scalable manner. We describe the production of CAR T cells from human peripheral blood mononuclear cells. We then detail a three-step staining protocol with metal-labeled antibodies and the subsequent mass cytometry analysis. This protocol allows simultaneous characterization of CAR T cell intracellular signaling, activation, proliferation, cytokine production, and phenotype in a single assay.

Published in STAR Protocols

ISSN: 2666-1667 (Online)
Publisher: Elsevier
Country of publisher: United States
LCC subjects: Science: Science (General)
Website: https://star-protocols.cell.com/

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Abstract

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