A robust pipeline for high-content, high-throughput immunophenotyping reveals age- and genetics-dependent changes in blood leukocytes.
CP: Immunology
CP: Systems biology
flow cytometry
high-dimensional flow cytometry
human immunology
immune aging
immune homeostasis
immune regulation
immunophenotyping studies
large cohorts
Journal
Cell reports methods
ISSN: 2667-2375
Titre abrégé: Cell Rep Methods
Pays: United States
ID NLM: 9918227360606676
Informations de publication
Date de publication:
23 Oct 2023
23 Oct 2023
Historique:
received:
20
09
2022
revised:
29
05
2023
accepted:
26
09
2023
medline:
30
10
2023
pubmed:
27
10
2023
entrez:
26
10
2023
Statut:
ppublish
Résumé
High-dimensional flow cytometry is the gold standard to study the human immune system in large cohorts. However, large sample sizes increase inter-experimental variation because of technical and experimental inaccuracies introduced by batch variability. Our high-throughput sample processing pipeline in combination with 28-color flow cytometry focuses on increased throughput (192 samples/experiment) and high reproducibility. We implemented quality control checkpoints to reduce technical and experimental variation. Finally, we integrated FlowSOM clustering to facilitate automated data analysis and demonstrate the reproducibility of our pipeline in a study with 3,357 samples. We reveal age-associated immune dynamics in 2,300 individuals, signified by decreasing T and B cell subsets with age. In addition, by combining genetic analyses, our approach revealed unique immune signatures associated with a single nucleotide polymorphism (SNP) that abrogates CD45 isoform splicing. In summary, we provide a versatile and reliable high-throughput, flow cytometry-based pipeline for immune discovery and exploration in large cohorts.
Identifiants
pubmed: 37883924
pii: S2667-2375(23)00283-7
doi: 10.1016/j.crmeth.2023.100619
pmc: PMC10626267
pii:
doi:
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
100619Informations de copyright
Published by Elsevier Inc.
Déclaration de conflit d'intérêts
Declaration of interests M.B. and M.R. are named as inventors on an NIH-filed patent for the Thawsome device. T.D.S. is co-founder of Zoe Ltd.
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