High-Content Immunophenotyping and Hierarchical Clustering Reveal Sources of Heterogeneity and New Surface Markers of Human Blood Monocyte Subsets
Language English Country Germany Media print-electronic
Document type Journal Article
Grant support
Ministry of Health of the Czech Republic
NV18-08-00385
Ministry of Education, Youth and Sports NPU I
LO1604
- MeSH
- Atherosclerosis diagnosis immunology MeSH
- Biodiversity MeSH
- Biomarkers metabolism MeSH
- Phenotype MeSH
- Immunophenotyping methods MeSH
- Blood Circulation MeSH
- Humans MeSH
- Lipopolysaccharide Receptors metabolism MeSH
- Monocytes physiology MeSH
- Flow Cytometry MeSH
- Receptors, IgG metabolism MeSH
- High-Throughput Screening Assays MeSH
- Cell Separation MeSH
- Cluster Analysis MeSH
- Inflammation diagnosis immunology MeSH
- Check Tag
- Humans MeSH
- Publication type
- Journal Article MeSH
- Names of Substances
- Biomarkers MeSH
- Lipopolysaccharide Receptors MeSH
- Receptors, IgG MeSH
OBJECTIVE: Blood monocyte subsets are emerging as biomarkers of cardiovascular inflammation. However, our understanding of human monocyte heterogeneity and their immunophenotypic features under healthy and inflammatory conditions is still evolving. RATIONALE: In this study, we sought to investigate the immunophenome of circulating human monocyte subsets. METHODS: Multiplexed, high-throughput flow cytometry screening arrays and computational data analysis were used to analyze the expression and hierarchical relationships of 242 specific surface markers on circulating classical (CD14++CD16-), intermediate (CD14++CD16+), and nonclassical (CD14+CD16++) monocytes in healthy adults. RESULTS: Using generalized linear models and hierarchical cluster analysis, we selected and clustered epitopes that most reliably differentiate between monocyte subsets. We validated existing transcriptional profiling data and revealed potential new surface markers that uniquely define the classical (e.g., BLTR1, CD35, CD38, CD49e, CD89, CD96), intermediate (e.g., CD39, CD275, CD305, CDw328), and nonclassical (e.g., CD29, CD132) subsets. In addition, our analysis revealed phenotypic cell clusters, identified by dendritic markers CMRF-44 and CMRF-56, independent of the traditional monocyte classification. CONCLUSION: These results reveal an advancement of the clinically applicable multiplexed screening arrays that may facilitate monocyte subset characterization and cytometry-based biomarker selection in various inflammatory disorders.
CLIP Childhood Leukemia Investigation Prague Charles University Prague Czech Republic
Department of Cardiology Kerckhoff Heart and Thorax Center Bad Nauheim Germany
Department of Cardiology St Johannes Hospital Dortmund Dortmund Germany
Department of Medicine 3 Cardiology Goethe University Hospital Frankfurt am Main Germany
German Center for Cardiovascular Research Partner site Rhine Main Frankfurt am Main Germany
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