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

Autor: Vitor Botafogo, Martín Pérez-Andres, María Jara-Acevedo, Paloma Bárcena, Georgiana Grigore, Alejandro Hernández-Delgado, Daniela Damasceno, Suzanne Comans, Elena Blanco, Alfonso Romero, Sonia Arriba-Méndez, Irene Gastaca-Abasolo, Carlos Eduardo Pedreira, Jacqueline A. M. van Gaans-van den Brink, Véronique Corbiere, Françoise Mascart, Cécile A. C. M. van Els, Alex-Mikael Barkoff, Andrea Mayado, Jacques J. M. van Dongen, Julia Almeida, Alberto Orfao
Přispěvatelé: Innovative Medicines Initiative, European Commission, European Federation of Pharmaceutical Industries and Associations, Bill & Melinda Gates Foundation, Instituto de Salud Carlos III, Fundación Mutua Madrileña, Ministerio de Ciencia, Innovación y Universidades (España), Conselho Nacional de Desenvolvimento Científico e Tecnológico (Brasil), Ministério da Ciência, Tecnologia e Inovação (Brasil), Agencia Estatal de Investigación (España)
Jazyk: angličtina
Rok vydání: 2020
Předmět:
0301 basic medicine
CD4-Positive T-Lymphocytes
Male
Lymphocytosis
Blood Donors
TFH
T-Lymphocytes
Regulatory
0302 clinical medicine
EuroFlow
Immunologie
Immunology and Allergy
Systemic mastocytosis
CD4+ T-cell subsets
Child
Original Research
Aged
80 and over
medicine.diagnostic_test
T-Lymphocytes
Helper-Inducer
Middle Aged
Fetal Blood
3. Good health
Phenotype
Cord blood
Child
Preschool
Female
medicine.symptom
Antibody
lcsh:Immunologic diseases. Allergy
Adult
immune monitoring
Adolescent
Immunology
Tregs
Biology
Flow cytometry
Immunophenotyping
03 medical and health sciences
Young Adult
Age Distribution
Monitoring
Immunologic
medicine
Humans
Aged
CD4+T-cell subsets
Common variable immunodeficiency
flow cytometry
Infant
Reproducibility of Results
medicine.disease
Sciences biomédicales
Th-cell subsets
Gene expression profiling
030104 developmental biology
biology.protein
lcsh:RC581-607
age-related values
030215 immunology
Zdroj: Frontiers in immunology, 11
Frontiers in Immunology, 11. FRONTIERS MEDIA SA
Frontiers in Immunology
Frontiers in Immunology, Vol 11 (2020)
Digital.CSIC. Repositorio Institucional del CSIC
instname
Popis: On behalf of the EuroFlow and PERISCOPE consortia.
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.
This work has been mainly supported by the PERISCOPE project, financed by the Innovative Medicines Initiative 2 Joint Undertaking (grant number 115910). This joint undertaking receives support from the European Union's Horizon 2020 Research and Innovation Programme, the European Federation of Pharmaceutical Industries and Associations (EFPIA), and The Bill and Melinda Gates Foundation (BMGF). The coordination and innovation processes of this study were supported by the EuroFlow Consortium. The EuroFlow Consortium received support from the FP6-2004-LIFESCIHEALTH-5 program of the European Commission (grant LSHB-CT-2006-018708) as Specific Targeted Research Project (STREP). Also, the study was partially supported by grants CB16/12/00400-FEDER [Biomedical Research Networking Center Consortium (CIBER-CIBERONC)], PI17/00399-FEDER, and PI19/01166-FEDER, Instituto de Salud Carlos III (ISCIII), Ministerio de Ciencia, Innovación y Universidades (Madrid, Spain), and a grant from Fundación Mutua Madrileña (Madrid, Spain). VB was supported by a grant from the Conselho Nacional de Desenvolvimento Científico e Tecnológico (grant number 207555/2015-0; Ministério da Ciência, Tecnologia, Inovações e Comunicações, Brazil). AH-D was supported by a grant from the Agencia Estatal de Investigación (grant number DI-17-09591, Ministerio de Ciencia, Innovación y Universidades, Spain).
Databáze: OpenAIRE
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