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

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

Research output: Contribution to journalArticle

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 workflowby 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.

Original languageEnglish (US)
Pages (from-to)1748-1758
Number of pages11
JournalJournal of Immunology
Volume198
Issue number4
DOIs
StatePublished - Feb 15 2017

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology

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    Burel, J. G., Qian, Y., Arlehamn, C. L., Weiskopf, D., Zapardiel-Gonzalo, J., Taplitz, R., Gilman, R. H., Saito, M., De Silva, A. D., Vijayanand, P., Scheuermann, R. H., Sette, A., & Peters, B. (2017). An integrated workflow to assess technical and biological variability of cell population frequencies in human peripheral blood by flow cytometry. Journal of Immunology, 198(4), 1748-1758. https://doi.org/10.4049/jimmunol.1601750