Novel Combination of Surface Markers for the Reliable and Comprehensive Identification of Human Thymic Epithelial Cells by Flow Cytometry: Quantitation and Transcriptional Characterization of Thymic Stroma in a Pediatric Cohort
| Autor: | Mathias Hauri-Hohl, Hitendu Dave, Maria Domenica Moccia, Veronika Haunerdinger, Stefano Vavassori, Lennart Opitz |
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| Přispěvatelé: | University of Zurich, Hauri-Hohl, Mathias M |
| Jazyk: | angličtina |
| Rok vydání: | 2021 |
| Předmět: |
Male
Chromosomes Human Pair 22 Cell Separation Transcriptome Extracellular matrix Human thymus Thymic epithelial cells Podoplanin CD49f CD200 Flow cytometry Immunology and Allergy Child PDPN Original Research human thymus medicine.diagnostic_test Age Factors Phenotype Cell biology Child Preschool thymic epithelial cells 2723 Immunology and Allergy Female Chromosome Deletion 22q11 Deletion Syndrome Stromal cell Adolescent extracellular matrix Immunology 610 Medicine & health 10071 Functional Genomics Center Zurich Thymus Gland Biology Sex Factors Myasthenia Gravis medicine Humans 2403 Immunology Gene Expression Profiling flow cytometry Mesenchymal stem cell Infant Newborn Infant Epithelial Cells RC581-607 podoplanin 10036 Medical Clinic 570 Life sciences biology Stromal Cells Immunologic diseases. Allergy Biomarkers |
| Zdroj: | Frontiers in Immunology, Vol 12 (2021) Frontiers in Immunology Frontiers in Immunology, 12 |
| ISSN: | 1664-3224 |
| Popis: | Thymic epithelial cells (TECs) are essential in supporting the development of mature T cells from hematopoietic progenitor cells and facilitate their lineage-commitment, proliferation, T-cell receptor repertoire selection and maturation. While animal model systems have greatly aided in elucidating the contribution of stromal cells to these intricate processes, human tissue has been more difficult to study, partly due to a lack of suitable surface markers comprehensively defining human TECs. Here, we conducted a flow cytometry based surface marker screen to reliably identify and quantify human TECs and delineate medullary from cortical subsets. These findings were validated by transcriptomic and histologic means. The combination of EpCAM, podoplanin (pdpn), CD49f and CD200 comprehensively identified human TECs and not only allowed their reliable distinction in medullary and cortical subsets but also their detailed quantitation. Transcriptomic profiling of each subset in comparison to fibroblasts and endothelial cells confirmed the identity of the different stromal cell subsets sorted according to the proposed strategy. Our dataset not only demonstrated transcriptional similarities between TEC and cells of mesenchymal origin but furthermore revealed a subset-specific distribution of a specific set of extracellular matrix-related genes in TECs. This indicates that TECs significantly contribute to the distinct compartmentalization – and thus function – of the human thymus. We applied the strategy to quantify TEC subsets in 31 immunologically healthy children, which revealed sex-specific differences of TEC composition early in life. As the distribution of mature CD4- or CD8-single-positive thymocytes was correspondingly altered, the composition of the thymic epithelial compartment may directly impact on the CD4-CD8-lineage choice of thymocytes. We prove that the plain, reliable strategy proposed here to comprehensively identify human TEC subpopulations by flow cytometry based on surface marker expression is suitable to determine their frequency and phenotype in health and disease and allows sorting of live cells for downstream analysis. Its use reaches from a reliable diagnostic tool for thymic biopsies to improved phenotypic characterization of thymic grafts intended for therapeutic use. Frontiers in Immunology, 12 ISSN:1664-3224 |
| Databáze: | OpenAIRE |
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