{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,10]],"date-time":"2026-10-10T04:39:10Z","timestamp":1791607150106,"version":"4.3.4"},"reference-count":33,"publisher":"Oxford University Press (OUP)","issue":"4","license":[{"start":{"date-parts":[[2017,2,1]],"date-time":"2017-02-01T00:00:00Z","timestamp":1485907200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/academic.oup.com\/pages\/standard-publication-reuse-rights"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2017,2,15]]},"abstract":"<jats:title>Abstract<\/jats:title>\n               <jats:p>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.<\/jats:p>","DOI":"10.4049\/jimmunol.1601750","type":"journal-article","created":{"date-parts":[[2017,1,10]],"date-time":"2017-01-10T02:20:22Z","timestamp":1484014822000},"page":"1748-1758","source":"Crossref","is-referenced-by-count":88,"title":["An Integrated Workflow To Assess Technical and Biological Variability of Cell Population Frequencies in Human Peripheral Blood by Flow Cytometry"],"prefix":"10.1093","volume":"198","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-1692-2758","authenticated-orcid":false,"given":"Julie G","family":"Burel","sequence":"first","affiliation":[{"name":"La Jolla Institute for Allergy and Immunology , La Jolla, CA 92037"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yu","family":"Qian","sequence":"additional","affiliation":[{"name":"J. Craig Venter Institute , La Jolla, CA 92037"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Cecilia","family":"Lindestam Arlehamn","sequence":"additional","affiliation":[{"name":"La Jolla Institute for Allergy and Immunology , La Jolla, CA 92037"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Daniela","family":"Weiskopf","sequence":"additional","affiliation":[{"name":"La Jolla Institute for Allergy and Immunology , La Jolla, CA 92037"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-9098-1652","authenticated-orcid":false,"given":"Jose","family":"Zapardiel-Gonzalo","sequence":"additional","affiliation":[{"name":"La Jolla Institute for Allergy and Immunology , La Jolla, CA 92037"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Randy","family":"Taplitz","sequence":"additional","affiliation":[{"name":"Division of Infectious Diseases, University of California , San Diego, La Jolla, CA 92093"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Robert H","family":"Gilman","sequence":"additional","affiliation":[{"name":"Johns Hopkins University Bloomberg School of Public Health , Baltimore, MD 21205"},{"name":"Universidad Peruana Caytano Hereida , Lima 15102,","place":["Peru"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mayuko","family":"Saito","sequence":"additional","affiliation":[{"name":"Johns Hopkins University Bloomberg School of Public Health , Baltimore, MD 21205"},{"name":"Universidad Peruana Caytano Hereida , Lima 15102,","place":["Peru"]},{"name":"Department of Virology, Tohoku University , Sendai 9808575,","place":["Japan"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Aruna D","family":"de Silva","sequence":"additional","affiliation":[{"name":"Genetech Research Center , Colombo 00800,","place":["Sri Lanka"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Pandurangan","family":"Vijayanand","sequence":"additional","affiliation":[{"name":"La Jolla Institute for Allergy and Immunology , La Jolla, CA 92037"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1355-892X","authenticated-orcid":false,"given":"Richard H","family":"Scheuermann","sequence":"additional","affiliation":[{"name":"La Jolla Institute for Allergy and Immunology , La Jolla, CA 92037"},{"name":"J. Craig Venter Institute , La Jolla, CA 92037"},{"name":"Department of Pathology, University of California , San Diego, La Jolla, CA 92093"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alessandro","family":"Sette","sequence":"additional","affiliation":[{"name":"La Jolla Institute for Allergy and Immunology , La Jolla, CA 92037"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8457-6693","authenticated-orcid":false,"given":"Bjoern","family":"Peters","sequence":"additional","affiliation":[{"name":"La Jolla Institute for Allergy and Immunology , La Jolla, CA 92037"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"286","published-online":{"date-parts":[[2017,2,15]]},"reference":[{"key":"2025030615030029500_r1","doi-asserted-by":"crossref","first-page":"77","DOI":"10.1016\/S0022-1759(00)00229-5","article-title":"A practical approach to multicolor flow cytometry for immunophenotyping","volume":"243","author":"Baumgarth","year":"2000","journal-title":"J. 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