Epigenetic immune cell counting in human blood samples for immunodiagnostics
| Autor: | Nina Babel, Manfred Gossen, Tina Baldinger, Dirk Schürmann, Ulrich Sack, David K. Stevenson, Ronald J. Wong, Udo Baron, Janika Schulze, Rosa Bacchetta, Jeannette Werner, Uwe-Gerd Liebert, Sven Olek, Andreas Grützkau, Carsten Speckmann, Konstantin Schildknecht, Stephan Borte, Andargaschew Mulu |
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| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
Cell Count HIV Infections Immunologic Tests Biology Epigenesis Genetic Flow cytometry Cohort Studies 03 medical and health sciences Neonatal Screening 0302 clinical medicine Immune system T-Lymphocyte Subsets medicine Humans Sulfites Whole blood Dried Blood Spot Testing Immunodiagnostics medicine.diagnostic_test Infant Newborn FOXP3 General Medicine DNA Methylation Cell counting 030104 developmental biology Genetic Loci 030220 oncology & carcinogenesis Immunology CD8 |
| Zdroj: | Science Translational Medicine. 10 |
| ISSN: | 1946-6242 1946-6234 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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