Mass cytometry deep phenotyping of human mononuclear phagocytes and myeloid-derived suppressor cells from human blood and bone marrow
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The monocyte phagocyte system (MPS) includes numerous monocyte, macrophage, and dendritic cell (DC) populations that are heterogeneous, both phenotypically and functionally. In this study, we sought to characterize those diverse MPS phenotypes with mass cytometry (CyTOF). To identify a deep phenotype of monocytes, macrophages, and DCs, a panel was designed to measure 38 identity, activation, and polarization markers, including CD14, CD16, HLA-DR, CD163, CD206, CD33, CD36, CD32, CD64, CD13, CD11b, CD11c, CD86, and CD274. MPS diversity was characterized for 1) circulating monocytes from healthy donors, 2) monocyte-derived macrophages further polarized in vitro (i.e., M-CSF, GM-CSF, IL-4, IL-10, IFN-γ, or LPS long-term stimulations), 3) monocyte-derived DCs, and 4) myeloid-derived suppressor cells (MDSCs), generated in vitro from bone marrow and/or peripheral blood. Known monocyte subsets were detected in peripheral blood to validate the panel and analysis pipeline. Then, using various culture conditions and stimuli before CyTOF analysis, we constructed a multidimensional framework for the MPS compartment, which was registered against historical M1 or M2 macrophages, monocyte subsets, and DCs. Notably, MDSCs generated in vitro from bone marrow expressed more S100A9 than when generated from peripheral blood. Finally, to test the approach in vivo, peripheral blood from patients with melanoma (n = 5) was characterized and observed to be enriched for MDSCs with a phenotype of CD14+HLA-DRlowS100A9high (3% of PBMCs in healthy donors, 15.5% in patients with melanoma, P < 0.02). In summary, mass cytometry comprehensively characterized phenotypes of human monocyte, MDSC, macrophage, and DC subpopulations in both in vitro models and patients.
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Exposure to supernatant from VIS cells induced macrophage activation marked by an M1-like polarization profile, CD86 upregulation, and signs of secondary macrophage senescence detected by SA-β-gal staining and senescence-typical gene expression.
“it was demonstrated that exposure to the supernatant of VIS cells induced macrophage activation that was marked by an M1-like polarization profile79,89, CD86 upregulation and signs of secondary macrophage senescence as detected by SA-β-gal staining and senescence-typical gene expression.”
From COVID-19 and cellular senescence (2023) · cites this paper as [79]
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- Authors
- Roussel, M., Ferrell, P. B., Greenplate, A. R., Lhomme, F., Le Gallou, S., Diggins, K. E., Johnson, D. B., & Irish, J. M.
- Affiliations
- Department of Pathology, Microbiology, and Immunology, Vanderbilt University School of Medicine , Nashville, Tennessee, USA
- Vanderbilt University (US)
- Centre Hospitalier Universitaire de Rennes (FR)
- CHU de Rennes, Pole de Biologie , Rennes, France
- Corresponding author
- Irish, Jonathan M
- ORCID
- Venue
- Journal of Leukocyte Biology
- Publisher
- Oxford University Press (OUP)
- Published
- 2017-04-11
- Volume
- 102
- Issue
- 2
- Pages
- 437-447
- ISSN
- 0741-5400 (ISSN-L)
- 1938-3673
- PMC
- PMC6608074
- OpenAlex
- W2607206500
- Cited by
- 93
- References
- 67
- Citations by year
- 2026: 4 · 2025: 7 · 2024: 7 · 2023: 12 · 2022: 12 · 2021: 16 · 2020: 14 · 2019: 11 · 2018: 8 · 2017: 2
- FWCI
- 4.33
- Type
- journal article
- Language
- en
- Open access
- green
- www.ncbi.nlm.nih.gov
- Licence
- academic.oup.com/journals/pages/open_access/funder_policies/chorus/standard_publication_model (vor)
- Locations
- Journal of Leukocyte Biology (publishedVersion)
- Europe PMC (PubMed Central) (publishedVersion)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- HAL (submittedVersion, other-oa)
- Funders
- Nuovo-Soldati Foundation (F31 CA199993, K12 CA090625, R25 CA136440-04, R00 CA143231-03)
- Vanderbilt-Ingram Cancer Center (P30 CA68485)
- Topics
- Life Sciences › Immunology and Microbiology › Immunology › Immune cells in cancer
- Life Sciences › Immunology and Microbiology › Immunology › Immunotherapy and Immune Responses
- Life Sciences › Immunology and Microbiology › Immunology › Immune responses and vaccinations
- Keywords
- monocytes, macrophages, dendritic cells, myeloid-derived suppressor cells, monocyte-derived macrophages, monocyte-derived dendritic cells, macrophage polarization, M1/M2 macrophages, melanoma, S100A9, CD14, HLA-DR, peripheral blood mononuclear cells, deep phenotyping
- MeSH terms
- Myeloid-Derived Suppressor Cells, Dendritic Cells, Flow Cytometry, Humans, Lymphocyte Culture Test, Mixed, Macrophages, Monocytes, Phagocytes, Phenotype
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