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dc.contributor.authorBotafogo, Vitor
dc.contributor.authorPérez-Andres, Martín
dc.contributor.authorJara-Acevedo, María
dc.contributor.authorBárcena, Paloma
dc.contributor.authorGrigore, Georgiana
dc.contributor.authorHernández-Delgado, Alejandro
dc.contributor.authorDamasceno, Daniela
dc.contributor.authorComans, Suzanne
dc.contributor.authorBlanco, Elena
dc.contributor.authorRomero, Alfonso
dc.contributor.authorArriba-Méndez, Sonia
dc.contributor.authorGastaca-Abasolo, Irene
dc.contributor.authorPedreira, Carlos Eduardo
dc.contributor.authorvan Gaans-van den Brink, Jacqueline A M
dc.contributor.authorCorbiere, Véronique
dc.contributor.authorMascart, Françoise
dc.contributor.authorvan Els, Cécile A C M
dc.contributor.authorBarkoff, Alex-Mikael
dc.contributor.authorMayado, Andrea
dc.contributor.authorvan Dongen, Jacques J M
dc.contributor.authorAlmeida, Julia
dc.contributor.authorOrfao, Alberto
dc.date.accessioned2020-08-19T14:27:17Z
dc.date.available2020-08-19T14:27:17Z
dc.date.issued2020-01-01
dc.identifier.issn1664-3224
dc.identifier.pmid32174910
dc.identifier.doi10.3389/fimmu.2020.00166
dc.identifier.urihttp://hdl.handle.net/10029/624157
dc.description.abstractCD4+ 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.en_US
dc.language.isoenen_US
dc.subjectCD4+ T-cell subsetsen_US
dc.subjectTFHen_US
dc.subjectTh-cell subsetsen_US
dc.subjectTregsen_US
dc.subjectage-related valuesen_US
dc.subjectflow cytometryen_US
dc.subjectimmune monitoringen_US
dc.titleAge 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_US
dc.typeArticleen_US
dc.identifier.journalFront Immunol 2020; 11:166en_US
dc.source.journaltitleFrontiers in immunology


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