A standardized flow cytometry procedure for the monitoring of regulatory T cells in clinical trials

Cytometry. Part B, Clinical Cytometry
Fabien PitoisetMichelle Rosenzwajg

Abstract

Quantification of regulatory T cells (Tregs) is crucial in immunomonitoring in clinical trials as this cell population has been shown to be involved in a wide range of diseases, including cancers, autoimmune diseases, infections, and allergies. Human Tregs are defined as CD4+ CD25+ CD127low FoxP3+ cells, and the standardization of their staining by flow cytometry is a challenge, especially in multicenter clinical trials, notably because of the intracellular location of FoxP3. A flow cytometry staining procedure was settled and standardized to measure human Tregs in peripheral whole blood using precoated dried antibodies in ready-to-use tubes. It was compared with reference methods and implemented and validated to be suitable with different cytometer platforms. The standardized protocol developed with dried antibodies and reduced volumes of whole blood allows an optimal identification of Tregs. Compared with classical staining procedure, it reduces the number of steps required, in a very fast and simple technique. The accuracy of the method was confirmed by a multicenter comparison with different cytometer brands. Our results highlight the reliability of this high-standard protocol that could become a reference method for the mo...Continue Reading

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Citations

Sep 1, 2018·Cytometry. Part a : the Journal of the International Society for Analytical Cytology·Fabien PitoisetMichelle Rosenzwajg
Sep 22, 2018·Cytometry. Part B, Clinical Cytometry·Sa A Wang
Oct 22, 2019·Cytometry. Part a : the Journal of the International Society for Analytical Cytology·Dawid SwiebodaFiona J Culley
Nov 1, 2018·PloS One·Yang ShenWolfgang Losert
Feb 18, 2020·Future Microbiology·Pénélope BourgoinThibaut Markarian
Aug 9, 2020·The Journal of Experimental Medicine·Carla Noemi CastroSeiamak Bahram
Mar 20, 2019·Cytometry. Part a : the Journal of the International Society for Analytical Cytology·Guillaume MonneretFabienne Venet
Apr 24, 2020·Journal of Immunology Research·Iole MacchiaFrancesca Urbani
Mar 7, 2021·Cells·Timoteo MarchiniDennis Wolf
May 21, 2021·Frontiers in Immunology·Ulrich BlacheUlrike Koehl

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