Thromb Haemost 2020; 120(01): 141-155
DOI: 10.1055/s-0039-1700871
New Technologies, Diagnostic Tools and Drugs
Georg Thieme Verlag KG Stuttgart · New York

High-Content Immunophenotyping and Hierarchical Clustering Reveal Sources of Heterogeneity and New Surface Markers of Human Blood Monocyte Subsets

Jedrzej Hoffmann*
1  Department of Medicine III, Cardiology, Goethe University Hospital, Frankfurt am Main, Germany
2  German Center for Cardiovascular Research (DZHK), Partner site Rhine-Main, Frankfurt am Main, Germany
,
Karel Fišer*
3  CLIP-Childhood Leukemia Investigation Prague, Charles University, Prague, Czech Republic
4  Department of Pediatric Hematology and Oncology, 2nd Faculty of Medicine, Charles University and Motol University Hospital, Prague, Czech Republic
,
Christoph Liebetrau
2  German Center for Cardiovascular Research (DZHK), Partner site Rhine-Main, Frankfurt am Main, Germany
5  Department of Cardiology, Kerckhoff Heart and Thorax Center, Bad Nauheim, Germany
,
Nora Staubach
5  Department of Cardiology, Kerckhoff Heart and Thorax Center, Bad Nauheim, Germany
,
David Kost
5  Department of Cardiology, Kerckhoff Heart and Thorax Center, Bad Nauheim, Germany
,
Sandra Voss
2  German Center for Cardiovascular Research (DZHK), Partner site Rhine-Main, Frankfurt am Main, Germany
5  Department of Cardiology, Kerckhoff Heart and Thorax Center, Bad Nauheim, Germany
,
Annkathrin zur Heiden
5  Department of Cardiology, Kerckhoff Heart and Thorax Center, Bad Nauheim, Germany
,
Oliver Dörr
2  German Center for Cardiovascular Research (DZHK), Partner site Rhine-Main, Frankfurt am Main, Germany
5  Department of Cardiology, Kerckhoff Heart and Thorax Center, Bad Nauheim, Germany
6  Division of Cardiology, Department of Internal Medicine I, University Hospital Giessen and Marburg, Giessen, Germany
,
Christoph Lipps
2  German Center for Cardiovascular Research (DZHK), Partner site Rhine-Main, Frankfurt am Main, Germany
5  Department of Cardiology, Kerckhoff Heart and Thorax Center, Bad Nauheim, Germany
6  Division of Cardiology, Department of Internal Medicine I, University Hospital Giessen and Marburg, Giessen, Germany
,
Holger M. Nef
2  German Center for Cardiovascular Research (DZHK), Partner site Rhine-Main, Frankfurt am Main, Germany
5  Department of Cardiology, Kerckhoff Heart and Thorax Center, Bad Nauheim, Germany
6  Division of Cardiology, Department of Internal Medicine I, University Hospital Giessen and Marburg, Giessen, Germany
,
Helge Möllmann
5  Department of Cardiology, Kerckhoff Heart and Thorax Center, Bad Nauheim, Germany
7  Department of Cardiology, St.-Johannes-Hospital Dortmund, Dortmund, Germany
,
Christian W. Hamm
2  German Center for Cardiovascular Research (DZHK), Partner site Rhine-Main, Frankfurt am Main, Germany
5  Department of Cardiology, Kerckhoff Heart and Thorax Center, Bad Nauheim, Germany
6  Division of Cardiology, Department of Internal Medicine I, University Hospital Giessen and Marburg, Giessen, Germany
,
Till Keller
2  German Center for Cardiovascular Research (DZHK), Partner site Rhine-Main, Frankfurt am Main, Germany
5  Department of Cardiology, Kerckhoff Heart and Thorax Center, Bad Nauheim, Germany
,
Christian Troidl
2  German Center for Cardiovascular Research (DZHK), Partner site Rhine-Main, Frankfurt am Main, Germany
5  Department of Cardiology, Kerckhoff Heart and Thorax Center, Bad Nauheim, Germany
6  Division of Cardiology, Department of Internal Medicine I, University Hospital Giessen and Marburg, Giessen, Germany
› Author Affiliations
Funding This study was supported by the Kerckhoff Institute for Heart Research (KHFI), Kerckhoff Foundation (Kerckhoff-Stiftung), and the German Centre for Cardiovascular Disease (DZHK). K.F. was supported by Ministry of Health of the Czech Republic (Grant Number: NV18–08–00385) and by Ministry of Education, Youth and Sports NPU I (Grant Number: LO1604).
Further Information

Publication History

05 February 2019

10 September 2019

Publication Date:
30 December 2019 (online)

Abstract

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.

* These authors contributed equally to this article.


Supplementary Material