Format

Send to

Choose Destination
Sci Transl Med. 2018 Aug 1;10(452). pii: eaan3508. doi: 10.1126/scitranslmed.aan3508.

Epigenetic immune cell counting in human blood samples for immunodiagnostics.

Author information

1
Ivana Türbachova Laboratory for Epigenetics, Epiontis GmbH, Precision for Medicine Group, 12489 Berlin, Germany.
2
Institute of Virology, Faculty of Medicine, University Leipzig, 04009 Leipzig, Germany.
3
Armauer Hansen Research Institute, 1005 Addis Ababa, Ethiopia.
4
Institute of Clinical Immunology, Faculty of Medicine, University Leipzig, 04009 Leipzig, Germany.
5
Center for Chronic Immunodeficiency and Department of Pediatric and Adolescent Medicine, Division of Pediatric Hematology and Oncology, Faculty of Medicine, University of Freiburg, 79110 Freiburg, Germany.
6
Institute of Biomaterial Science, Helmholtz-Zentrum Geesthacht, 14513 Teltow, Germany.
7
Berlin-Brandenburg Center for Regenerative Therapies, 13353 Berlin, Germany.
8
Division of Neonatal and Developmental Medicine, Department of Pediatrics, Stanford University School of Medicine, Stanford, CA 94305, USA.
9
Marienhospital Herne, Medizinische Klinik I, Universität Bochum, 44625 Herne, Germany.
10
Charité Universitätsmedizin Berlin, 13353 Berlin, Germany.
11
Division of Stem Cell Transplantation and Regenerative Medicine, Department of Pediatrics, Stanford School of Medicine, Stanford, CA 94305, USA.
12
Deutsches Rheuma-Forschungszentrum, an Institute of the Leibniz Association, Immune Monitoring Core Facility, 10117 Berlin, Germany.
13
ImmunoDeficiencyCenter Leipzig, Municipal Hospital St. Georg Leipzig, 04129 Leipzig, Germany. sven.olek@epiontis.com stephan.borte@idcl.de.
14
Division of Clinical Immunology, Department of Laboratory Medicine, Karolinska University Hospital Huddinge at Karolinska Institutet, 14186 Stockholm, Sweden.
15
Ivana Türbachova Laboratory for Epigenetics, Epiontis GmbH, Precision for Medicine Group, 12489 Berlin, Germany. sven.olek@epiontis.com stephan.borte@idcl.de.

Abstract

Immune cell profiles provide valuable diagnostic information for hematologic and immunologic diseases. Although it is the most widely applied analytical approach, flow cytometry is limited to liquid blood. Moreover, either analysis must be performed with fresh samples or cell integrity needs to be guaranteed during storage and transport. We developed epigenetic real-time quantitative polymerase chain reaction (qPCR) assays for analysis of human leukocyte subpopulations. After method establishment, whole blood from 25 healthy donors and 97 HIV+ patients as well as dried spots from 250 healthy newborns and 24 newborns with primary immunodeficiencies were analyzed. Concordance between flow cytometric and epigenetic data for neutrophils and B, natural killer, CD3+ T, CD8+ T, CD4+ T, and FOXP3+ regulatory T cells was evaluated, demonstrating substantial equivalence between epigenetic qPCR analysis and flow cytometry. Epigenetic qPCR achieves both relative and absolute quantifications. Applied to dried blood spots, epigenetic immune cell quantification was shown to identify newborns suffering from various primary immunodeficiencies. Using epigenetic qPCR not only provides a precise means for immune cell counting in fresh-frozen blood but also extends applicability to dried blood spots. This method could expand the ability for screening immune defects and facilitates diagnostics of unobservantly collected samples, for example, in underdeveloped areas, where logistics are major barriers to screening.

Supplemental Content

Full text links

Icon for HighWire
Loading ...
Support Center