A standardized flow cytometry procedure ...
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Article dans une revue scientifique
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Title :
A standardized flow cytometry procedure for the monitoring of regulatory T cells in clinical trials.
Author(s) :
Pitoiset, Fabien [Auteur]
Immunologie - Immunopathologie - Immunothérapie [I3]
Barbie, Michele [Auteur]
Immunologie - Immunopathologie - Immunothérapie [I3]
Monneret, Guillaume [Auteur]
Braudeau, Cecile [Auteur]
Centre de Recherche en Transplantation et Immunologie [U1064 Inserm - CRTI]
Pochard, Pierre [Auteur]
Immunologie et Pathologie [EA2216]
Pellegrin, Isabelle [Auteur]
Trauet, Jacques [Auteur]
Lille Inflammation Research International Center - U 995 [LIRIC]
Labalette, Myriam [Auteur]
Lille Inflammation Research International Center - U 995 [LIRIC]
Klatzmann, David [Auteur]
Immunologie - Immunopathologie - Immunothérapeutique [I3]
Rosenzwajg, Michelle [Auteur]
Immunologie - Immunopathologie - Immunothérapeutique [I3]
Immunologie - Immunopathologie - Immunothérapie [I3]
Barbie, Michele [Auteur]
Immunologie - Immunopathologie - Immunothérapie [I3]
Monneret, Guillaume [Auteur]
Braudeau, Cecile [Auteur]
Centre de Recherche en Transplantation et Immunologie [U1064 Inserm - CRTI]
Pochard, Pierre [Auteur]
Immunologie et Pathologie [EA2216]
Pellegrin, Isabelle [Auteur]
Trauet, Jacques [Auteur]

Lille Inflammation Research International Center - U 995 [LIRIC]
Labalette, Myriam [Auteur]

Lille Inflammation Research International Center - U 995 [LIRIC]
Klatzmann, David [Auteur]
Immunologie - Immunopathologie - Immunothérapeutique [I3]
Rosenzwajg, Michelle [Auteur]
Immunologie - Immunopathologie - Immunothérapeutique [I3]
Journal title :
Cytometry Part B: Clinical Cytometry
Abbreviated title :
Cytometry B Clin Cytom
Volume number :
94
Pages :
777-782
Publication date :
2018-09
Keyword(s) :
flow cytometry
regulatory T cells
standardization
regulatory T cells
standardization
HAL domain(s) :
Sciences du Vivant [q-bio]
English abstract : [en]
Background: 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 ...
Show more >Background: 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. Method: 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. Results: 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. Conclusions: Our results highlight the reliability of this high-standard protocol that could become a reference method for the monitoring of Tregs in clinical trials. © 2018 International Clinical Cytometry SocietyShow less >
Show more >Background: 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. Method: 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. Results: 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. Conclusions: Our results highlight the reliability of this high-standard protocol that could become a reference method for the monitoring of Tregs in clinical trials. © 2018 International Clinical Cytometry SocietyShow less >
Language :
Anglais
Audience :
Internationale
Popular science :
Non
Administrative institution(s) :
Inserm
Université de Lille
CHU Lille
Université de Lille
CHU Lille
Research team(s) :
Immunity, inflammation and fibrsis in auto and allo-reactivity
Submission date :
2019-03-01T14:25:46Z
2024-02-02T09:27:18Z
2024-02-02T09:27:18Z
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