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An Integrated Workflow To Assess Technical and Biological Variability of Cell Population Frequencies in Human Peripheral Blood by Flow Cytometry
- Source :
- The Journal of Immunology. 198:1748-1758
- Publication Year :
- 2017
- Publisher :
- The American Association of Immunologists, 2017.
-
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.
- Subjects :
- Adult
CD4-Positive T-Lymphocytes
Male
0301 basic medicine
T cell
Immunology
Population
Cell Count
Context (language use)
Biology
Article
Immunophenotyping
Workflow
Flow cytometry
03 medical and health sciences
Sex Factors
0302 clinical medicine
Immune system
Latent Tuberculosis
T-Lymphocyte Subsets
medicine
Humans
Immunology and Allergy
education
Systems immunology
education.field_of_study
Latent tuberculosis
medicine.diagnostic_test
Age Factors
Replicate
Flow Cytometry
medicine.disease
CD56 Antigen
030104 developmental biology
medicine.anatomical_structure
Leukocytes, Mononuclear
Female
030215 immunology
Subjects
Details
- ISSN :
- 15506606 and 00221767
- Volume :
- 198
- Database :
- OpenAIRE
- Journal :
- The Journal of Immunology
- Accession number :
- edsair.doi.dedup.....57007949ddf1d582902ad5cea8dc0d14