Mass Cytometry: a robust platform for the comprehensive immunomonitoring of CAR‐T‐cell therapies

Corneau, Aurélien; Parizot, Christophe; Cheraï, Mustapha; Todesco, Eve; Blanc, Catherine; Литвинова, Е. Г.; Nguyen, Stéphanie; Roos‐Weil, Damien; Guihot, Amélie; Norol, Françoise

doi:10.1111/bjh.17551

Cited by 6 publications

(2 citation statements)

References 15 publications

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“…The mass cytometry-related protocol can be applied to clinical CAR T cell products for extensive characterization. For this purpose, we recommend the inclusion in the antibody panel of one metal-labeled antibody that specifically recognizes the CAR T cells (e.g., anti-biotin CD19Fc), as proposed by ( Corneau et al., 2021 ).…”

Section: Before You Beginmentioning

confidence: 99%

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

et al. 2022

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Section: Before You Beginmentioning

confidence: 99%

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

et al. 2022

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“…However, implementation of multi‐color flow cytometric analysis for PK in a multicenter clinical trial presents several logistical challenges and low sensitivity 11,12 . Advanced mass cytometric technologies (cytometry by time of flight) are very useful in immunophenotyping, immuno‐monitoring, and persistence of CAR‐T‐cells, 13–15 but are not widely used to measure CAR‐T transgene levels due to challenges similar to that of flow cytometry. The PCR‐based amplification of CAR genes as a measure of PK has become prevalent due to its ability to amplify the signal even from a few cells and the ease of sample processing from frozen cells derived from whole blood or enriched T‐cells 16,17 .…”

mentioning

confidence: 99%

Bioanalytical Methods for Characterization of CAR‐T Cellular Kinetics: Comparison of PCR Assays and Matrices

Masilamani

Jawa

Dai

et al. 2023

Clin Pharma and Therapeutics

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Recently, multiple chimeric antigen receptor T‐cell (CAR‐T)‐based therapies have been approved for treating hematological malignancies, targeting CD19 and B‐cell maturation antigen. Unlike protein or antibody therapies, CAR‐T therapies are “living cell” therapies whose pharmacokinetics are characterized by expansion, distribution, contraction, and persistence. Therefore, this unique modality requires a different approach for quantitation compared with conventional ligand binding assays implemented for most biologics. Cellular (flow cytometry) or molecular assays (polymerase chain reaction (PCR)) can be deployed with each having unique advantages and disadvantages. In this article, we describe the molecular assays utilized: quantitative PCR (qPCR), which was the initial platform used to estimate transgene copy numbers and more recently droplet digital PCR (ddPCR) which quantitates the absolute copy numbers of CAR transgene. The comparability of the two methods in patient samples and of each method across different matrices (isolated CD3+ T‐cells or whole blood) was also performed. The results show a good correlation between qPCR and ddPCR for the amplification of same gene in clinical samples from a CAR‐T therapy trial. In addition, our studies show that the qPCR‐based amplification of transgene levels was well‐correlated, independent of DNA sources (either CD3+ T‐cells or whole blood). Our results also highlight that ddPCR can be a better platform for monitoring samples at the early phase of CAR‐T dosing prior to expansion and during long‐term monitoring as they can detect samples with very low copy numbers with high sensitivity, in addition to easier implementation and sample logistics.

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Successful treatment by CAR T-cells in multi-refractory mantle cell lymphoma with central nervous system involvement

Caillet,

Houillier,

Sourdeau

et al. 2023

Ann Hematol

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Mass Cytometry: a robust platform for the comprehensive immunomonitoring of CAR‐T‐cell therapies

Cited by 6 publications

References 15 publications

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

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

Bioanalytical Methods for Characterization of CAR‐T Cellular Kinetics: Comparison of PCR Assays and Matrices

Successful treatment by CAR T-cells in multi-refractory mantle cell lymphoma with central nervous system involvement

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