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

Antibodies Humans Leukocytes, Mononuclear T-Lymphocytes
Cell Biology Cell culture Cell isolation Flow Cytometry/Mass Cytometry Health Sciences Immunology Single Cell

Journal

STAR protocols
ISSN: 2666-1667
Titre abrégé: STAR Protoc
Pays: United States
ID NLM: 101769501

Informations de publication

Date de publication:
18 03 2022
Historique:
entrez: 24 2 2022
pubmed: 25 2 2022
medline: 9 4 2022
Statut: epublish

Résumé

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.

Identifiants

DOI: 10.1016/j.xpro.2022.101174 PMID: 35199038 PMC: PMC8844283
pubmed: 35199038
doi: 10.1016/j.xpro.2022.101174
pii: S2666-1667(22)00054-5
pmc: PMC8844283
doi:

Substances chimiques

Antibodies 0

Types de publication

Journal Article Research Support, Non-U.S. Gov't

Langues

eng

Sous-ensembles de citation

IM

Pagination

101174

Subventions

Organisme : Department of Health
Pays : United Kingdom

Informations de copyright

© 2022 The Author(s).

Déclaration de conflit d'intérêts

T.A.A. is currently an employee of Fluidigm.

Références

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Auteurs

Ilaria M Michelozzi (IM)

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.

Jahangir Sufi (J)

Cell Communication Lab, Department of Oncology, University College London Cancer Institute, WC1E 6DD London, UK.

Thomas A Adejumo (TA)

Fluidigm Corporation, South San Francisco, CA, USA.

Persis J Amrolia (PJ)

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 (CJ)

Cell Communication Lab, Department of Oncology, University College London Cancer Institute, WC1E 6DD London, UK.

Alice Giustacchini (A)

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.

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Classifications MeSH

questionsmedicales.fr › Acides aminés, peptides et protéines › Protéines › Globulines › Sérum-globulines › Immunoglobulines › Anticorps › High-dimensional functional phenotyping of...
questionsmedicales.fr › Eucaryotes › Animaux › Chordés › Vertébrés › Mammifères › Eutheria › Primates › Haplorhini › Catarrhini › Hominidae › Humains › High-dimensional functional phenotyping of...
questionsmedicales.fr › Systèmes sanguin et immunitaire › Système immunitaire › Leucocytes › Agranulocytes › High-dimensional functional phenotyping of...
questionsmedicales.fr › Systèmes sanguin et immunitaire › Système immunitaire › Leucocytes › Agranulocytes › Lymphocytes › Lymphocytes T › High-dimensional functional phenotyping of...