title: Age and gender leucocytes variances and references values generated using the standardized ONE-Study protocol.
abstract: Flow cytometry is now accepted as an ideal technology to reveal changes in immune cell composition and function. However, it is also an error-prone and variable technology, which makes it difficult to reproduce findings across laboratories. We have recently developed a strategy to standardize whole blood flow cytometry. The performance of our protocols was challenged here by profiling samples from healthy volunteers to reveal age- and gender-dependent differences and to establish a standardized reference cohort for use in clinical trials. Whole blood samples from two different cohorts were analyzed (first cohort: n = 52, second cohort: n = 46, both 20-84 years with equal gender distribution). The second cohort was run as a validation cohort by a different operator. The "ONE Study" panels were applied to analyze expression of >30 different surface markers to enumerate proportional and absolute numbers of >50 leucocyte subsets. Indeed, analysis of the first cohort revealed significant age-dependent changes in subsets e.g. increased activated and differentiated CD4(+) and CD8(+) T cell subsets, acquisition of a memory phenotype for Tregs as well as decreased MDC2 and Marginal Zone B cells. Males and females showed different dynamics in age-dependent T cell activation and differentiation, indicating faster immunosenescence in males. Importantly, although both cohorts consisted of a small sample size, our standardized approach enabled validation of age-dependent changes with the second cohort. Thus, we have proven the utility of our strategy and generated reproducible reference ranges accounting for age- and gender-dependent differences, which are crucial for a better patient monitoring and individualized therapy. © 2016 International Society for Advancement of Cytometry.
authors.affiliation: Institute of Medical Immunology, Charité Universitätsmedizin Berlin, Germany.|Department of International Health, Immunology and Microbiology, Faculty of Health and Medical Sciences, University of Copenhagen, København, Denmark.
authors.forename: Anders H
authors.initials: AH
authors.lastname: Kverneland
authors.affiliation: Institute of Medical Immunology, Charité Universitätsmedizin Berlin, Germany.
authors.forename: Mathias
authors.initials: M
authors.lastname: Streitz
authors.affiliation: Department of Surgery, University of Regensburg, Germany.
authors.forename: Edward
authors.initials: E
authors.lastname: Geissler
authors.affiliation: Department of Surgery, University of Regensburg, Germany.
authors.forename: James
authors.initials: J
authors.lastname: Hutchinson
authors.affiliation: Institute of Medical Immunology, Charité Universitätsmedizin Berlin, Germany.
authors.forename: Katrin
authors.initials: K
authors.lastname: Vogt
authors.affiliation: Institute of Medical Immunology, Charité Universitätsmedizin Berlin, Germany.
authors.forename: David
authors.initials: D
authors.lastname: Boës
authors.affiliation: Institute of Medical Immunology, Charité Universitätsmedizin Berlin, Germany.
authors.forename: Nadja
authors.initials: N
authors.lastname: Niemann
authors.affiliation: Department of International Health, Immunology and Microbiology, Faculty of Health and Medical Sciences, University of Copenhagen, København, Denmark.
authors.forename: Anders Elm
authors.initials: AE
authors.lastname: Pedersen
authors.affiliation: Institute of Medical Immunology, Charité Universitätsmedizin Berlin, Germany.
authors.forename: Stephan
authors.initials: S
authors.lastname: Schlickeiser
authors.affiliation: Institute of Medical Immunology, Charité Universitätsmedizin Berlin, Germany.
authors.forename: Birgit
authors.initials: B
authors.lastname: Sawitzki
mesh_terms: D000328:Adult; D000367:Age Factors; D000368:Aged; D000369:Aged, 80 and over; D015703:Antigens, CD / Q000235:genetics / Q000276:immunology*; D015331:Cohort Studies; D005260:Female; D005434:Flow Cytometry / Q000592:standards*; D064368:Healthy Volunteers; D006801:Humans; D007156:Immunologic Memory; D016130:Immunophenotyping / Q000592:standards*; D016131:Lymphocyte Subsets / Q000145:classification* / Q000166:cytology / Q000276:immunology; D008297:Male; D008875:Middle Aged; D012016:Reference Values; D012737:Sex Factors
chemical_list: D015703:Antigens, CD
keywords: adaptive immunity; aging; immunosenescence; innate immunity; standardization