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An integrated workflow to assess technical and biological variability of cell population frequencies in human peripheral blood by flow cytometry

An integrated workflow to assess technical and biological variability of cell population frequencies in human peripheral blood by flow cytometry
An integrated workflow to assess technical and biological variability of cell population frequencies in human peripheral blood by flow cytometry

In the context of large-scale human system immunology studies, controlling for technical and biological variability is crucial to ensure that experimental data support research conclusions. In this study, we report on a universal workflow to evaluate both technical and biological variation in multiparameter flow cytometry, applied to the development of a 10-color panel to identify all major cell populations and T cell subsets in cryopreserved PBMC. Replicate runs from a control donation and comparison of different gating strategies assessed the technical variability associated with each cell population and permitted the calculation of a quality control score. Applying our panel to a large collection of PBMC samples, we found that most cell populations showed low intraindividual variability over time. In contrast, certain subpopulations such as CD56 T cells and Temra CD4 T cells were associated with high interindividual variability. Age but not gender had a significant effect on the frequency of several populations, with a drastic decrease in naive T cells observed in older donors. Ethnicity also influenced a significant proportion of immune cell population frequencies, emphasizing the need to account for these covariates in immune profiling studies. We also exemplify the usefulness of our workflow by identifying a novel cell-subset signature of latent tuberculosis infection. Thus, our study provides a universal workflow to establish and evaluate any flow cytometry panel in systems immunology studies.

Adult, Age Factors, CD4-Positive T-Lymphocytes, CD56 Antigen, Cell Count, Female, Flow Cytometry, Humans, Immunophenotyping, Latent Tuberculosis, Leukocytes, Mononuclear, Male, Sex Factors, T-Lymphocyte Subsets, Workflow, Journal Article, Research Support, N.I.H., Extramural
0022-1767
1748-1758
Burel, Julie G.
d043b967-6296-4f02-b628-e397a82de132
Qian, Yu
edb8082b-c264-46e4-beae-8e35ceb3e035
Lindestam Arlehamn, Cecilia
972358b3-c46c-4de5-b7e1-afa2cf65ed35
Weiskopf, Daniela
0baaee36-7c36-49e1-a97e-9251d8d34904
Zapardiel-Gonzalo, Jose
700ca752-a4e5-430c-9f8d-e903a197cd79
Taplitz, Randy
2d810857-e357-49ea-949e-ea6b7c42cb9d
Gilman, Robert H
d948b282-8e0c-43de-aeda-4b8c8a789870
Saito, Mayuko
de3e8fc4-9321-4fab-8f4e-1d8660cf7ae3
de Silva, Aruna D
408fe904-2994-49c8-bbcd-5091551924c4
Vijayanand, Pandurangan
79514f33-66cf-47cc-a8fa-46bbfc21b7d1
Scheuermann, Richard H
50df1335-f518-4a86-b948-2e8eee8ad3b7
Sette, Alessandro
240988b3-3c05-4041-a39b-8be8e145803c
Peters, Bjoern
c49863c8-d2c6-4db4-9d2c-72a7c0cbe117
Burel, Julie G.
d043b967-6296-4f02-b628-e397a82de132
Qian, Yu
edb8082b-c264-46e4-beae-8e35ceb3e035
Lindestam Arlehamn, Cecilia
972358b3-c46c-4de5-b7e1-afa2cf65ed35
Weiskopf, Daniela
0baaee36-7c36-49e1-a97e-9251d8d34904
Zapardiel-Gonzalo, Jose
700ca752-a4e5-430c-9f8d-e903a197cd79
Taplitz, Randy
2d810857-e357-49ea-949e-ea6b7c42cb9d
Gilman, Robert H
d948b282-8e0c-43de-aeda-4b8c8a789870
Saito, Mayuko
de3e8fc4-9321-4fab-8f4e-1d8660cf7ae3
de Silva, Aruna D
408fe904-2994-49c8-bbcd-5091551924c4
Vijayanand, Pandurangan
79514f33-66cf-47cc-a8fa-46bbfc21b7d1
Scheuermann, Richard H
50df1335-f518-4a86-b948-2e8eee8ad3b7
Sette, Alessandro
240988b3-3c05-4041-a39b-8be8e145803c
Peters, Bjoern
c49863c8-d2c6-4db4-9d2c-72a7c0cbe117

Burel, Julie G., Qian, Yu, Lindestam Arlehamn, Cecilia, Weiskopf, Daniela, Zapardiel-Gonzalo, Jose, Taplitz, Randy, Gilman, Robert H, Saito, Mayuko, de Silva, Aruna D, Vijayanand, Pandurangan, Scheuermann, Richard H, Sette, Alessandro and Peters, Bjoern (2017) An integrated workflow to assess technical and biological variability of cell population frequencies in human peripheral blood by flow cytometry. The Journal of Immunology, 198 (4), 1748-1758. (doi:10.4049/jimmunol.1601750).

Record type: Article

Abstract

In the context of large-scale human system immunology studies, controlling for technical and biological variability is crucial to ensure that experimental data support research conclusions. In this study, we report on a universal workflow to evaluate both technical and biological variation in multiparameter flow cytometry, applied to the development of a 10-color panel to identify all major cell populations and T cell subsets in cryopreserved PBMC. Replicate runs from a control donation and comparison of different gating strategies assessed the technical variability associated with each cell population and permitted the calculation of a quality control score. Applying our panel to a large collection of PBMC samples, we found that most cell populations showed low intraindividual variability over time. In contrast, certain subpopulations such as CD56 T cells and Temra CD4 T cells were associated with high interindividual variability. Age but not gender had a significant effect on the frequency of several populations, with a drastic decrease in naive T cells observed in older donors. Ethnicity also influenced a significant proportion of immune cell population frequencies, emphasizing the need to account for these covariates in immune profiling studies. We also exemplify the usefulness of our workflow by identifying a novel cell-subset signature of latent tuberculosis infection. Thus, our study provides a universal workflow to establish and evaluate any flow cytometry panel in systems immunology studies.

Full text not available from this repository.

More information

Accepted/In Press date: 6 December 2016
e-pub ahead of print date: 31 January 2017
Published date: 15 February 2017
Additional Information: Copyright © 2017 by The American Association of Immunologists, Inc.
Keywords: Adult, Age Factors, CD4-Positive T-Lymphocytes, CD56 Antigen, Cell Count, Female, Flow Cytometry, Humans, Immunophenotyping, Latent Tuberculosis, Leukocytes, Mononuclear, Male, Sex Factors, T-Lymphocyte Subsets, Workflow, Journal Article, Research Support, N.I.H., Extramural

Identifiers

Local EPrints ID: 422753
URI: https://eprints.soton.ac.uk/id/eprint/422753
ISSN: 0022-1767
PURE UUID: d017466a-64dd-486f-bd60-b2aa038bb1ce

Catalogue record

Date deposited: 03 Aug 2018 16:30
Last modified: 13 Mar 2019 18:14

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Contributors

Author: Julie G. Burel
Author: Yu Qian
Author: Cecilia Lindestam Arlehamn
Author: Daniela Weiskopf
Author: Jose Zapardiel-Gonzalo
Author: Randy Taplitz
Author: Robert H Gilman
Author: Mayuko Saito
Author: Aruna D de Silva
Author: Pandurangan Vijayanand
Author: Richard H Scheuermann
Author: Alessandro Sette
Author: Bjoern Peters

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