Age Distribution of Multiple Functionally Relevant Subsets of CD4+ T Cells in Human Blood Using a Standardized and Validated 14-Color EuroFlow Immune Monitoring Tube
[EN]CD4+ T cells comprise multiple functionally distinct cell populations that play a key role in immunity. Despite blood monitoring of CD4+ T-cell subsets is of potential clinical utility, no standardized and validated approaches have been proposed so far. The aim of this study was to design and va...
| Autores: | , , , , , , , , , , , , , , , , , , , , , |
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| Tipo de recurso: | artículo |
| Estado: | Versión publicada |
| Fecha de publicación: | 2020 |
| País: | España |
| Institución: | Universidad de Salamanca (USAL) |
| Repositorio: | GREDOS. Repositorio Institucional de la Universidad de Salamanca |
| OAI Identifier: | oai:gredos.usal.es:10366/168638 |
| Acceso en línea: | http://hdl.handle.net/10366/168638 |
| Access Level: | acceso abierto |
| Palabra clave: | Immunomonitoring CD4+ T-cell populations Flow cytometry T-Lymphocyte Subsets Flow Cytometry CD4-Positive T-Lymphocytes linfocitos T CD4-positivos subgrupos de linfocitos T citometría de flujo |
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Age Distribution of Multiple Functionally Relevant Subsets of CD4+ T Cells in Human Blood Using a Standardized and Validated 14-Color EuroFlow Immune Monitoring TubeBotafogo Goncalves, VitorPérez Andrés, MartínJara Acevedo, MaríaBárcena Carrasco, PalomaGrigore, GeorgianaHernández Delgado, AlejandroPinto Damasceno, DanielaComans, SuzanneBlanco Álvarez, ElenaRomero, AlfonsoArriba Méndez, Sonia deGastaca Abasolo, IrenePedreira, Carlos Eduardovan Gaans-van den Brink, Jacqueline A. M.Corbiere, VéroniqueMascart, Françoisevan Els, Cécile A. C. M.Barkoff, Alex-MikaelMayado, Andreavan Dongen, Jacques J. M.Almeida Parra, Julia MaríaOrfao de Matos Correia e Vale, José AlbertoImmunomonitoringCD4+ T-cell populationsFlow cytometryT-Lymphocyte SubsetsFlow CytometryCD4-Positive T-Lymphocyteslinfocitos T CD4-positivossubgrupos de linfocitos Tcitometría de flujo[EN]CD4+ T cells comprise multiple functionally distinct cell populations that play a key role in immunity. Despite blood monitoring of CD4+ T-cell subsets is of potential clinical utility, no standardized and validated approaches have been proposed so far. The aim of this study was to design and validate a single 14-color antibody combination for sensitive and reproducible flow cytometry monitoring of CD4+ T-cell populations in human blood to establish normal age-related reference values and evaluate the presence of potentially altered profiles in three distinct disease models-monoclonal B-cell lymphocytosis (MBL), systemic mastocytosis (SM), and common variable immunodeficiency (CVID). Overall, 145 blood samples from healthy donors were used to design and validate a 14-color antibody combination based on extensive reagent testing in multiple cycles of design-testing-evaluation-redesign, combined with in vitro functional studies, gene expression profiling, and multicentric evaluation of manual vs. automated gating. Fifteen cord blood and 98 blood samples from healthy donors (aged 0-89 years) were used to establish reference values, and another 25 blood samples were evaluated for detecting potentially altered CD4 T-cell subset profiles in MBL (n = 8), SM (n = 7), and CVID (n = 10). The 14-color tube can identify ≥89 different CD4+ T-cell populations in blood, as validated with high multicenter reproducibility, particularly when software-guided automated (vs. manual expert-based) gating was used. Furthermore, age-related reference values were established, which reflect different kinetics for distinct subsets: progressive increase of naïve T cells, T-helper (Th)1, Th17, follicular helper T (TFH) cells, and regulatory T cells (Tregs) from birth until 2 years, followed by a decrease of naïve T cells, Th2, and Tregs in older children and a subsequent increase in multiple Th-cell subsets toward late adulthood. Altered and unique CD4+ T-cell subset profiles were detected in two of the three disease models evaluated (SM and CVID). In summary, the EuroFlow immune monitoring TCD4 tube allows fast, automated, and reproducible identification of ≥89 subsets of CD4+ blood T cells, with different kinetics throughout life. These results set the basis for in-depth T-cell monitoring in different disease and therapeutic conditions.Frontiers Media202620262020info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionapplication/pdfhttp://hdl.handle.net/10366/168638reponame:GREDOS. Repositorio Institucional de la Universidad de Salamancainstname:Universidad de Salamanca (USAL)InglésPI17/00399PI19/01166CB16/12/00400Attribution 4.0 Internacionalhttp://creativecommons.org/licenses/by/4.0/info:eu-repo/semantics/openAccessoai:gredos.usal.es:10366/1686382026-06-07T06:28:51Z |
| dc.title.none.fl_str_mv |
Age Distribution of Multiple Functionally Relevant Subsets of CD4+ T Cells in Human Blood Using a Standardized and Validated 14-Color EuroFlow Immune Monitoring Tube |
| title |
Age Distribution of Multiple Functionally Relevant Subsets of CD4+ T Cells in Human Blood Using a Standardized and Validated 14-Color EuroFlow Immune Monitoring Tube |
| spellingShingle |
Age Distribution of Multiple Functionally Relevant Subsets of CD4+ T Cells in Human Blood Using a Standardized and Validated 14-Color EuroFlow Immune Monitoring Tube Botafogo Goncalves, Vitor Immunomonitoring CD4+ T-cell populations Flow cytometry T-Lymphocyte Subsets Flow Cytometry CD4-Positive T-Lymphocytes linfocitos T CD4-positivos subgrupos de linfocitos T citometría de flujo |
| title_short |
Age Distribution of Multiple Functionally Relevant Subsets of CD4+ T Cells in Human Blood Using a Standardized and Validated 14-Color EuroFlow Immune Monitoring Tube |
| title_full |
Age Distribution of Multiple Functionally Relevant Subsets of CD4+ T Cells in Human Blood Using a Standardized and Validated 14-Color EuroFlow Immune Monitoring Tube |
| title_fullStr |
Age Distribution of Multiple Functionally Relevant Subsets of CD4+ T Cells in Human Blood Using a Standardized and Validated 14-Color EuroFlow Immune Monitoring Tube |
| title_full_unstemmed |
Age Distribution of Multiple Functionally Relevant Subsets of CD4+ T Cells in Human Blood Using a Standardized and Validated 14-Color EuroFlow Immune Monitoring Tube |
| title_sort |
Age Distribution of Multiple Functionally Relevant Subsets of CD4+ T Cells in Human Blood Using a Standardized and Validated 14-Color EuroFlow Immune Monitoring Tube |
| dc.creator.none.fl_str_mv |
Botafogo Goncalves, Vitor Pérez Andrés, Martín Jara Acevedo, María Bárcena Carrasco, Paloma Grigore, Georgiana Hernández Delgado, Alejandro Pinto Damasceno, Daniela Comans, Suzanne Blanco Álvarez, Elena Romero, Alfonso Arriba Méndez, Sonia de Gastaca Abasolo, Irene Pedreira, Carlos Eduardo van Gaans-van den Brink, Jacqueline A. M. Corbiere, Véronique Mascart, Françoise van Els, Cécile A. C. M. Barkoff, Alex-Mikael Mayado, Andrea van Dongen, Jacques J. M. Almeida Parra, Julia María Orfao de Matos Correia e Vale, José Alberto |
| author |
Botafogo Goncalves, Vitor |
| author_facet |
Botafogo Goncalves, Vitor Pérez Andrés, Martín Jara Acevedo, María Bárcena Carrasco, Paloma Grigore, Georgiana Hernández Delgado, Alejandro Pinto Damasceno, Daniela Comans, Suzanne Blanco Álvarez, Elena Romero, Alfonso Arriba Méndez, Sonia de Gastaca Abasolo, Irene Pedreira, Carlos Eduardo van Gaans-van den Brink, Jacqueline A. M. Corbiere, Véronique Mascart, Françoise van Els, Cécile A. C. M. Barkoff, Alex-Mikael Mayado, Andrea van Dongen, Jacques J. M. Almeida Parra, Julia María Orfao de Matos Correia e Vale, José Alberto |
| author_role |
author |
| author2 |
Pérez Andrés, Martín Jara Acevedo, María Bárcena Carrasco, Paloma Grigore, Georgiana Hernández Delgado, Alejandro Pinto Damasceno, Daniela Comans, Suzanne Blanco Álvarez, Elena Romero, Alfonso Arriba Méndez, Sonia de Gastaca Abasolo, Irene Pedreira, Carlos Eduardo van Gaans-van den Brink, Jacqueline A. M. Corbiere, Véronique Mascart, Françoise van Els, Cécile A. C. M. Barkoff, Alex-Mikael Mayado, Andrea van Dongen, Jacques J. M. Almeida Parra, Julia María Orfao de Matos Correia e Vale, José Alberto |
| author2_role |
author author author author author author author author author author author author author author author author author author author author author |
| dc.subject.none.fl_str_mv |
Immunomonitoring CD4+ T-cell populations Flow cytometry T-Lymphocyte Subsets Flow Cytometry CD4-Positive T-Lymphocytes linfocitos T CD4-positivos subgrupos de linfocitos T citometría de flujo |
| topic |
Immunomonitoring CD4+ T-cell populations Flow cytometry T-Lymphocyte Subsets Flow Cytometry CD4-Positive T-Lymphocytes linfocitos T CD4-positivos subgrupos de linfocitos T citometría de flujo |
| description |
[EN]CD4+ T cells comprise multiple functionally distinct cell populations that play a key role in immunity. Despite blood monitoring of CD4+ T-cell subsets is of potential clinical utility, no standardized and validated approaches have been proposed so far. The aim of this study was to design and validate a single 14-color antibody combination for sensitive and reproducible flow cytometry monitoring of CD4+ T-cell populations in human blood to establish normal age-related reference values and evaluate the presence of potentially altered profiles in three distinct disease models-monoclonal B-cell lymphocytosis (MBL), systemic mastocytosis (SM), and common variable immunodeficiency (CVID). Overall, 145 blood samples from healthy donors were used to design and validate a 14-color antibody combination based on extensive reagent testing in multiple cycles of design-testing-evaluation-redesign, combined with in vitro functional studies, gene expression profiling, and multicentric evaluation of manual vs. automated gating. Fifteen cord blood and 98 blood samples from healthy donors (aged 0-89 years) were used to establish reference values, and another 25 blood samples were evaluated for detecting potentially altered CD4 T-cell subset profiles in MBL (n = 8), SM (n = 7), and CVID (n = 10). The 14-color tube can identify ≥89 different CD4+ T-cell populations in blood, as validated with high multicenter reproducibility, particularly when software-guided automated (vs. manual expert-based) gating was used. Furthermore, age-related reference values were established, which reflect different kinetics for distinct subsets: progressive increase of naïve T cells, T-helper (Th)1, Th17, follicular helper T (TFH) cells, and regulatory T cells (Tregs) from birth until 2 years, followed by a decrease of naïve T cells, Th2, and Tregs in older children and a subsequent increase in multiple Th-cell subsets toward late adulthood. Altered and unique CD4+ T-cell subset profiles were detected in two of the three disease models evaluated (SM and CVID). In summary, the EuroFlow immune monitoring TCD4 tube allows fast, automated, and reproducible identification of ≥89 subsets of CD4+ blood T cells, with different kinetics throughout life. These results set the basis for in-depth T-cell monitoring in different disease and therapeutic conditions. |
| publishDate |
2020 |
| dc.date.none.fl_str_mv |
2020 2026 2026 |
| dc.type.none.fl_str_mv |
info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion |
| format |
article |
| status_str |
publishedVersion |
| dc.identifier.none.fl_str_mv |
http://hdl.handle.net/10366/168638 |
| url |
http://hdl.handle.net/10366/168638 |
| dc.language.none.fl_str_mv |
Inglés |
| language_invalid_str_mv |
Inglés |
| dc.relation.none.fl_str_mv |
PI17/00399 PI19/01166 CB16/12/00400 |
| dc.rights.none.fl_str_mv |
Attribution 4.0 Internacional http://creativecommons.org/licenses/by/4.0/ info:eu-repo/semantics/openAccess |
| rights_invalid_str_mv |
Attribution 4.0 Internacional http://creativecommons.org/licenses/by/4.0/ |
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openAccess |
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application/pdf |
| dc.publisher.none.fl_str_mv |
Frontiers Media |
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Frontiers Media |
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reponame:GREDOS. Repositorio Institucional de la Universidad de Salamanca instname:Universidad de Salamanca (USAL) |
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Universidad de Salamanca (USAL) |
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GREDOS. Repositorio Institucional de la Universidad de Salamanca |
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GREDOS. Repositorio Institucional de la Universidad de Salamanca |
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15,811543 |