A flow cytometry approach reveals heterogeneity in conventional subsets of murine renal mononuclear phagocytes
Abstract Mononuclear phagocytes (MNPs) participate in inflammation and repair after kidney injury, reflecting their complex nature. Dissection into refined functional subunits has been challenging and would benefit understanding of renal pathologies. Flow cytometric approaches are limited to classif...
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| Format: | Article |
| Language: | English |
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Nature Portfolio
2021-06-01
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| Series: | Scientific Reports |
| Online Access: | https://doi.org/10.1038/s41598-021-92784-x |
| _version_ | 1818403152342286336 |
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| author | Johannes Nordlohne Ilona Hulsmann Svenja Schwafertz Jasmin Zgrajek Manuel Grundmann Sibylle von Vietinghoff Frank Eitner Michael S. Becker |
| author_facet | Johannes Nordlohne Ilona Hulsmann Svenja Schwafertz Jasmin Zgrajek Manuel Grundmann Sibylle von Vietinghoff Frank Eitner Michael S. Becker |
| author_sort | Johannes Nordlohne |
| collection | DOAJ |
| description | Abstract Mononuclear phagocytes (MNPs) participate in inflammation and repair after kidney injury, reflecting their complex nature. Dissection into refined functional subunits has been challenging and would benefit understanding of renal pathologies. Flow cytometric approaches are limited to classifications of either different MNP subsets or functional state. We sought to combine these two dimensions in one protocol that considers functional heterogeneity in each MNP subset. We identified five distinct renal MNP subsets based on a previously described strategy. In vitro polarization of bone marrow-derived macrophages (BMDM) into M1- and M2-like cells suggested functional distinction of CD86 + MHCII + CD206- and CD206 + cells. Combination of both distinction methods identified CD86 + MHCII + CD206- and CD206 + cells in all five MNP subsets, revealing their heterologous nature. Our approach revealed that MNP composition and their functional segmentation varied between different mouse models of kidney injury and, moreover, was dynamically regulated in a time-dependent manner. CD206 + cells from three analyzed MNP subsets had a higher ex vivo phagocytic capacity than CD86 + MHCII + CD206- counterparts, indicating functional uniqueness of each subset. In conclusion, our novel flow cytometric approach refines insights into renal MNP heterogeneity and therefore could benefit mechanistic understanding of renal pathology. |
| first_indexed | 2024-12-14T08:19:43Z |
| format | Article |
| id | doaj.art-9f7f1171fe0e426d9a83fd8ce3509300 |
| institution | Directory Open Access Journal |
| issn | 2045-2322 |
| language | English |
| last_indexed | 2024-12-14T08:19:43Z |
| publishDate | 2021-06-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Reports |
| spelling | doaj.art-9f7f1171fe0e426d9a83fd8ce35093002022-12-21T23:09:50ZengNature PortfolioScientific Reports2045-23222021-06-0111111510.1038/s41598-021-92784-xA flow cytometry approach reveals heterogeneity in conventional subsets of murine renal mononuclear phagocytesJohannes Nordlohne0Ilona Hulsmann1Svenja Schwafertz2Jasmin Zgrajek3Manuel Grundmann4Sibylle von Vietinghoff5Frank Eitner6Michael S. Becker7Cardiovascular Research, Research and Development, Pharmaceuticals, Kidney Diseases, Bayer AGCardiovascular Research, Research and Development, Pharmaceuticals, Kidney Diseases, Bayer AGCardiovascular Research, Research and Development, Pharmaceuticals, Kidney Diseases, Bayer AGCardiovascular Research, Research and Development, Pharmaceuticals, Kidney Diseases, Bayer AGCardiovascular Research, Research and Development, Pharmaceuticals, Kidney Diseases, Bayer AGNephrology Section, Medical Clinic 1, University Hospital Bonn, Rheinische Friedrich-Wilhelms UniversityCardiovascular Research, Research and Development, Pharmaceuticals, Kidney Diseases, Bayer AGCardiovascular Research, Research and Development, Pharmaceuticals, Kidney Diseases, Bayer AGAbstract Mononuclear phagocytes (MNPs) participate in inflammation and repair after kidney injury, reflecting their complex nature. Dissection into refined functional subunits has been challenging and would benefit understanding of renal pathologies. Flow cytometric approaches are limited to classifications of either different MNP subsets or functional state. We sought to combine these two dimensions in one protocol that considers functional heterogeneity in each MNP subset. We identified five distinct renal MNP subsets based on a previously described strategy. In vitro polarization of bone marrow-derived macrophages (BMDM) into M1- and M2-like cells suggested functional distinction of CD86 + MHCII + CD206- and CD206 + cells. Combination of both distinction methods identified CD86 + MHCII + CD206- and CD206 + cells in all five MNP subsets, revealing their heterologous nature. Our approach revealed that MNP composition and their functional segmentation varied between different mouse models of kidney injury and, moreover, was dynamically regulated in a time-dependent manner. CD206 + cells from three analyzed MNP subsets had a higher ex vivo phagocytic capacity than CD86 + MHCII + CD206- counterparts, indicating functional uniqueness of each subset. In conclusion, our novel flow cytometric approach refines insights into renal MNP heterogeneity and therefore could benefit mechanistic understanding of renal pathology.https://doi.org/10.1038/s41598-021-92784-x |
| spellingShingle | Johannes Nordlohne Ilona Hulsmann Svenja Schwafertz Jasmin Zgrajek Manuel Grundmann Sibylle von Vietinghoff Frank Eitner Michael S. Becker A flow cytometry approach reveals heterogeneity in conventional subsets of murine renal mononuclear phagocytes Scientific Reports |
| title | A flow cytometry approach reveals heterogeneity in conventional subsets of murine renal mononuclear phagocytes |
| title_full | A flow cytometry approach reveals heterogeneity in conventional subsets of murine renal mononuclear phagocytes |
| title_fullStr | A flow cytometry approach reveals heterogeneity in conventional subsets of murine renal mononuclear phagocytes |
| title_full_unstemmed | A flow cytometry approach reveals heterogeneity in conventional subsets of murine renal mononuclear phagocytes |
| title_short | A flow cytometry approach reveals heterogeneity in conventional subsets of murine renal mononuclear phagocytes |
| title_sort | flow cytometry approach reveals heterogeneity in conventional subsets of murine renal mononuclear phagocytes |
| url | https://doi.org/10.1038/s41598-021-92784-x |
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