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Our recent studies indicate that regulation of CD163 mRNA expression is a key functional property of polarized monocytes and macrophages and is mediated at the transcriptional and posttranscriptional level, including via microRNAs. The goal of the current study is to develop a multiparameter flow cytometry panel incorporating detection of CD163 mRNA for polarized monocyte and macrophage populations in disorders such as SJIA and MAS. THP-1 cells and CD14+ human monocytes were stained using fluorochrome-conjugated Abs to myeloid surface markers, along with CD163 mRNA. Staining for mRNA could reliably detect CD163 expression while simultaneously detecting different macrophage populations using Abs targeting CD14, CD64, CD80, CD163, and CD209. This approach was found to be highly sensitive for increased mRNA expression when macrophages were polarized with IL-10 [M(IL-10)], with a strong signal over a broad range of IL-10 concentrations, and showed distinct kinetics of CD163 mRNA and protein induction upon IL-10 stimulation. Finally, this panel demonstrated clear changes in polarization markers in unstimulated monocytes from patients with SJIA and MAS, including upregulated CD163 mRNA and increased CD64 expression. This approach represents a robust and sensitive system for RNA flow cytometry, useful for studying CD163 expression as part of a multimarker panel for human monocytes and macrophages, with broad applicability to the pathogenesis of hyperinflammatory diseases.<\/jats:p>","DOI":"10.4049\/jimmunol.1800765","type":"journal-article","created":{"date-parts":[[2019,1,25]],"date-time":"2019-01-25T20:39:28Z","timestamp":1548448768000},"page":"1635-1643","source":"Crossref","is-referenced-by-count":27,"title":["A Multiparameter Flow Cytometry Analysis Panel to Assess CD163 mRNA and Protein in Monocyte and Macrophage Populations in Hyperinflammatory Diseases"],"prefix":"10.1093","volume":"202","author":[{"given":"Sherry","family":"Thornton","sequence":"first","affiliation":[{"name":"Division of Rheumatology, Cincinnati Children\u2019s Hospital Medical Center, University of Cincinnati College of Medicine , Cincinnati, OH 45229"},{"name":"Department of Pediatrics, University of Cincinnati College of Medicine , Cincinnati, OH 45267"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rachel","family":"Tan","sequence":"additional","affiliation":[{"name":"Division of Rheumatology, Cincinnati Children\u2019s Hospital Medical Center, University of Cincinnati College of Medicine , Cincinnati, OH 45229"},{"name":"Department of Pediatrics, University of Cincinnati College of Medicine , Cincinnati, OH 45267"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alyssa","family":"Sproles","sequence":"additional","affiliation":[{"name":"Division of Rheumatology, Cincinnati Children\u2019s Hospital Medical Center, University of Cincinnati College of Medicine , Cincinnati, OH 45229"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Thuy","family":"Do","sequence":"additional","affiliation":[{"name":"Division of Rheumatology, Cincinnati Children\u2019s Hospital Medical Center, University of Cincinnati College of Medicine , Cincinnati, OH 45229"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jonathan","family":"Schick","sequence":"additional","affiliation":[{"name":"Division of Rheumatology, Cincinnati Children\u2019s Hospital Medical Center, University of Cincinnati College of Medicine , Cincinnati, OH 45229"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alexei A","family":"Grom","sequence":"additional","affiliation":[{"name":"Division of Rheumatology, Cincinnati Children\u2019s Hospital Medical Center, University of Cincinnati College of Medicine , Cincinnati, OH 45229"},{"name":"Department of Pediatrics, University of Cincinnati College of Medicine , Cincinnati, OH 45267"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Monica","family":"DeLay","sequence":"additional","affiliation":[{"name":"Division of Rheumatology, Cincinnati Children\u2019s Hospital Medical Center, University of Cincinnati College of Medicine , Cincinnati, OH 45229"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5923-7051","authenticated-orcid":false,"given":"Grant S","family":"Schulert","sequence":"additional","affiliation":[{"name":"Division of Rheumatology, Cincinnati Children\u2019s Hospital Medical Center, University of Cincinnati College of Medicine , Cincinnati, OH 45229"},{"name":"Department of Pediatrics, University of Cincinnati College of Medicine , Cincinnati, OH 45267"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"286","published-online":{"date-parts":[[2019,3,1]]},"reference":[{"key":"2025030417560835800_r1","doi-asserted-by":"crossref","first-page":"145","DOI":"10.1146\/annurev-med-061813-012806","article-title":"Pathogenesis of macrophage activation syndrome and potential for cytokine- directed therapies","volume":"66","author":"Schulert","year":"2015","journal-title":"Annu. 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