Advanced immunophenotyping of lymphocyte and monocyte subsets in healthy Australian adults using a novel spectral flow cytometry panel

IntroductionLymphocytes play pivotal roles in disease pathogenesis and can be used as potential biomarkers for various immunological conditions. Yet, current flow cytometry methods used in clinical settings are often only capable of measuring between four to eight distinct lymphocyte populations. Th...

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Main Authors: Ainsley R. Davies, Kristy Kwong, Zhijia Yu, Koula E. M. Diamand, Fei-Ju Li, Laurensia Kannitha, Sidra A. Ali, Abolfazl Amjadipour, Ann-Maree Padarin, Michael Devoy, Harpreet Vohra, Bahar Miraghazadeh, Simon H. Jiang, Anne Brüstle, Nicolas Cherbuin, Christopher J. Nolan, Matthew C. Cook, Elizabeth E. Gardiner, Stuart Read, Euan McNaughton, Katrina L. Randall
Format: Article
Language:English
Published: Frontiers Media S.A. 2025-07-01
Series:Frontiers in Immunology
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Online Access:https://www.frontiersin.org/articles/10.3389/fimmu.2025.1577206/full
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author Ainsley R. Davies
Ainsley R. Davies
Kristy Kwong
Kristy Kwong
Kristy Kwong
Zhijia Yu
Koula E. M. Diamand
Koula E. M. Diamand
Fei-Ju Li
Fei-Ju Li
Laurensia Kannitha
Sidra A. Ali
Abolfazl Amjadipour
Ann-Maree Padarin
Michael Devoy
Harpreet Vohra
Bahar Miraghazadeh
Simon H. Jiang
Simon H. Jiang
Anne Brüstle
Nicolas Cherbuin
Christopher J. Nolan
Christopher J. Nolan
Matthew C. Cook
Elizabeth E. Gardiner
Stuart Read
Stuart Read
Euan McNaughton
Katrina L. Randall
Katrina L. Randall
Katrina L. Randall
author_facet Ainsley R. Davies
Ainsley R. Davies
Kristy Kwong
Kristy Kwong
Kristy Kwong
Zhijia Yu
Koula E. M. Diamand
Koula E. M. Diamand
Fei-Ju Li
Fei-Ju Li
Laurensia Kannitha
Sidra A. Ali
Abolfazl Amjadipour
Ann-Maree Padarin
Michael Devoy
Harpreet Vohra
Bahar Miraghazadeh
Simon H. Jiang
Simon H. Jiang
Anne Brüstle
Nicolas Cherbuin
Christopher J. Nolan
Christopher J. Nolan
Matthew C. Cook
Elizabeth E. Gardiner
Stuart Read
Stuart Read
Euan McNaughton
Katrina L. Randall
Katrina L. Randall
Katrina L. Randall
author_sort Ainsley R. Davies
collection DOAJ
description IntroductionLymphocytes play pivotal roles in disease pathogenesis and can be used as potential biomarkers for various immunological conditions. Yet, current flow cytometry methods used in clinical settings are often only capable of measuring between four to eight distinct lymphocyte populations. The purpose of our study was to measure many lymphocyte and monocyte populations from a single sample, with the long-term aim of validating our assay for diagnostic use in the Australian regulatory environment.MethodsWe designed and optimised a novel 30-colour lymphocyte immunophenotyping panel tailored for use on a 3-laser (V-B-R) spectral flow cytometer. This panel measures over 50 lymphocyte and monocyte populations.ResultsIn this report we present data derived from 148 healthy individuals.DiscussionThis lays the groundwork for future clinical application of spectral flow cytometry tests and offers a more comprehensive approach to lymphocyte and monocyte analysis with future implications for disease diagnosis and monitoring.
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spelling doaj-art-5659ad155f2f45db8c227b210c0d48ef2025-08-20T02:50:20ZengFrontiers Media S.A.Frontiers in Immunology1664-32242025-07-011610.3389/fimmu.2025.15772061577206Advanced immunophenotyping of lymphocyte and monocyte subsets in healthy Australian adults using a novel spectral flow cytometry panelAinsley R. Davies0Ainsley R. Davies1Kristy Kwong2Kristy Kwong3Kristy Kwong4Zhijia Yu5Koula E. M. Diamand6Koula E. M. Diamand7Fei-Ju Li8Fei-Ju Li9Laurensia Kannitha10Sidra A. Ali11Abolfazl Amjadipour12Ann-Maree Padarin13Michael Devoy14Harpreet Vohra15Bahar Miraghazadeh16Simon H. Jiang17Simon H. Jiang18Anne Brüstle19Nicolas Cherbuin20Christopher J. Nolan21Christopher J. Nolan22Matthew C. Cook23Elizabeth E. Gardiner24Stuart Read25Stuart Read26Euan McNaughton27Katrina L. Randall28Katrina L. Randall29Katrina L. Randall30Canberra Clinical Phenomics Service, John Curtin School of Medical Research, The Australian National University, Canberra, ACT, AustraliaPhenomics Translation Initiative, John Curtin School of Medical Research, Australian Phenomics Facility, The Australian National University, Canberra, ACT, AustraliaCanberra Clinical Phenomics Service, John Curtin School of Medical Research, The Australian National University, Canberra, ACT, AustraliaPhenomics Translation Initiative, John Curtin School of Medical Research, Australian Phenomics Facility, The Australian National University, Canberra, ACT, AustraliaDepartment of Medicine, University of Cambridge, Cambridge, United KingdomCanberra Clinical Phenomics Service, John Curtin School of Medical Research, The Australian National University, Canberra, ACT, AustraliaPhenomics Translation Initiative, John Curtin School of Medical Research, Australian Phenomics Facility, The Australian National University, Canberra, ACT, AustraliaANU Centre for Therapeutic Discovery, John Curtin School of Medical Research, Australian Phenomics Facility, The Australian National University, Canberra, ACT, AustraliaPhenomics Translation Initiative, John Curtin School of Medical Research, Australian Phenomics Facility, The Australian National University, Canberra, ACT, AustraliaCytometry, Histology and Advanced Spatial Multiomics, John Curtin School of Medical Research, The Australian National University, Canberra, ACT, AustraliaCanberra Clinical Phenomics Service, John Curtin School of Medical Research, The Australian National University, Canberra, ACT, AustraliaDivision of Genome Science and Cancer, John Curtin School of Medical Research, The Australian National University, Canberra, ACT, AustraliaDivision of Immunology and Infectious Disease, John Curtin School of Medical Research, The Australian National University, Canberra, ACT, AustraliaCanberra Clinical Phenomics Service, John Curtin School of Medical Research, The Australian National University, Canberra, ACT, AustraliaCytometry, Histology and Advanced Spatial Multiomics, John Curtin School of Medical Research, The Australian National University, Canberra, ACT, AustraliaCytometry, Histology and Advanced Spatial Multiomics, John Curtin School of Medical Research, The Australian National University, Canberra, ACT, AustraliaDivision of Immunology and Infectious Disease, John Curtin School of Medical Research, The Australian National University, Canberra, ACT, AustraliaDivision of Immunology and Infectious Disease, John Curtin School of Medical Research, The Australian National University, Canberra, ACT, AustraliaDepartment of Renal Medicine, The Canberra Hospital, Canberra, ACT, AustraliaDivision of Immunology and Infectious Disease, John Curtin School of Medical Research, The Australian National University, Canberra, ACT, AustraliaNational Centre for Epidemiology and Population Health, Australian National University, Canberra, ACT, AustraliaDivision of Immunology and Infectious Disease, John Curtin School of Medical Research, The Australian National University, Canberra, ACT, Australia0School of Medicine and Psychology, The Australian National University, Canberra, ACT, AustraliaDepartment of Medicine, University of Cambridge, Cambridge, United KingdomDivision of Genome Science and Cancer, John Curtin School of Medical Research, The Australian National University, Canberra, ACT, AustraliaPhenomics Translation Initiative, John Curtin School of Medical Research, Australian Phenomics Facility, The Australian National University, Canberra, ACT, Australia1South Australian Health and Medical Research Institute, Adelaide, SA, AustraliaCanberra Clinical Phenomics Service, John Curtin School of Medical Research, The Australian National University, Canberra, ACT, AustraliaCanberra Clinical Phenomics Service, John Curtin School of Medical Research, The Australian National University, Canberra, ACT, Australia0School of Medicine and Psychology, The Australian National University, Canberra, ACT, Australia2Department of Immunology, and Department of Immunopathology, ACT Pathology, Canberra Health Services, Canberra, ACT, AustraliaIntroductionLymphocytes play pivotal roles in disease pathogenesis and can be used as potential biomarkers for various immunological conditions. Yet, current flow cytometry methods used in clinical settings are often only capable of measuring between four to eight distinct lymphocyte populations. The purpose of our study was to measure many lymphocyte and monocyte populations from a single sample, with the long-term aim of validating our assay for diagnostic use in the Australian regulatory environment.MethodsWe designed and optimised a novel 30-colour lymphocyte immunophenotyping panel tailored for use on a 3-laser (V-B-R) spectral flow cytometer. This panel measures over 50 lymphocyte and monocyte populations.ResultsIn this report we present data derived from 148 healthy individuals.DiscussionThis lays the groundwork for future clinical application of spectral flow cytometry tests and offers a more comprehensive approach to lymphocyte and monocyte analysis with future implications for disease diagnosis and monitoring.https://www.frontiersin.org/articles/10.3389/fimmu.2025.1577206/fullspectral flow cytometrylymphocytePBMCimmunophenotypingautoimmunityT cell
spellingShingle Ainsley R. Davies
Ainsley R. Davies
Kristy Kwong
Kristy Kwong
Kristy Kwong
Zhijia Yu
Koula E. M. Diamand
Koula E. M. Diamand
Fei-Ju Li
Fei-Ju Li
Laurensia Kannitha
Sidra A. Ali
Abolfazl Amjadipour
Ann-Maree Padarin
Michael Devoy
Harpreet Vohra
Bahar Miraghazadeh
Simon H. Jiang
Simon H. Jiang
Anne Brüstle
Nicolas Cherbuin
Christopher J. Nolan
Christopher J. Nolan
Matthew C. Cook
Elizabeth E. Gardiner
Stuart Read
Stuart Read
Euan McNaughton
Katrina L. Randall
Katrina L. Randall
Katrina L. Randall
Advanced immunophenotyping of lymphocyte and monocyte subsets in healthy Australian adults using a novel spectral flow cytometry panel
Frontiers in Immunology
spectral flow cytometry
lymphocyte
PBMC
immunophenotyping
autoimmunity
T cell
title Advanced immunophenotyping of lymphocyte and monocyte subsets in healthy Australian adults using a novel spectral flow cytometry panel
title_full Advanced immunophenotyping of lymphocyte and monocyte subsets in healthy Australian adults using a novel spectral flow cytometry panel
title_fullStr Advanced immunophenotyping of lymphocyte and monocyte subsets in healthy Australian adults using a novel spectral flow cytometry panel
title_full_unstemmed Advanced immunophenotyping of lymphocyte and monocyte subsets in healthy Australian adults using a novel spectral flow cytometry panel
title_short Advanced immunophenotyping of lymphocyte and monocyte subsets in healthy Australian adults using a novel spectral flow cytometry panel
title_sort advanced immunophenotyping of lymphocyte and monocyte subsets in healthy australian adults using a novel spectral flow cytometry panel
topic spectral flow cytometry
lymphocyte
PBMC
immunophenotyping
autoimmunity
T cell
url https://www.frontiersin.org/articles/10.3389/fimmu.2025.1577206/full
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