Spectral cytometry of rheumatoid arthritis patients implicates myeloid dendritic cells and granular HLA-DR+CD15+CD16+ cells in pro-inflammatory antigen presentation

IntroductionRheumatoid arthritis (RA) is a systemic autoimmune disease that leads to inflammation of synovial joints and other organs. Many RA patients “share” a common peptide sequence within the HLA-DR (MHC II) molecule expressed on antigen-presenting cells (APC), suggesting that HLA-DR+ cells are...

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Main Authors: Christian Geier, Haani Qudsi, Estelle Khairallah, Jihad Ben Gabr, Robert Winchester, Andras Perl
Format: Article
Language:English
Published: Frontiers Media S.A. 2025-07-01
Series:Frontiers in Immunology
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Online Access:https://www.frontiersin.org/articles/10.3389/fimmu.2025.1596609/full
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author Christian Geier
Haani Qudsi
Estelle Khairallah
Jihad Ben Gabr
Robert Winchester
Andras Perl
author_facet Christian Geier
Haani Qudsi
Estelle Khairallah
Jihad Ben Gabr
Robert Winchester
Andras Perl
author_sort Christian Geier
collection DOAJ
description IntroductionRheumatoid arthritis (RA) is a systemic autoimmune disease that leads to inflammation of synovial joints and other organs. Many RA patients “share” a common peptide sequence within the HLA-DR (MHC II) molecule expressed on antigen-presenting cells (APC), suggesting that HLA-DR+ cells are important in RA inflammation. We use HLA-DR positivity to comprehensively immunophenotype APC by spectral cytometry.MethodsWe measured mean fluorescence intensities (MFI) of HLA-DR and molecules associated with dendritic cells (CD141, CD1c, CD163, CD11c, CD123, and CD303), monocytes (CD14 and CD16), granulocytic markers (CD15 and CCR3), co-stimulatory molecules (CD86 and CD275), and chemokine receptors (CCR2, CCR3, and CCR7) from RA patients and healthy donors by spectral flow cytometry.ResultsDC2 (CD1c+) showed higher CD86, CD275 (ICOS-L), CD56, and CCR7 in RA (all p < 0.05). CD56 was also increased in (CD163+) DC3 (p = 0.0453). CD15 was increased throughout RA dendritic cell subsets and classical and intermediate monocytes (all p < 0.01). Except for B cells, HLA-DR was not different in RA. A distinct HLA-DR+CD15+CD16+ population appeared in RA (p = 0.0004), which contributed a mean of 1.3% (± SD 2.85%) to the overall HLA-DR+ APC compartment. This HLA-DR+CD15+CD16+ subset was positive for CD83, CD275, and, like plasmacytoid pDC, CD303+. However, in contrast to pDC, it formed distinct t-SNE clusters and differed from reference pDC (CD123+CD303+) by much less CD123 (p < 0.01). The HLA-DR+CD15+CD16+ phenotype correlated with clinical markers of systemic inflammation (p < 0.01).DiscussionIn conclusion, dendritic cell and monocyte alterations in RA include an increased co-stimulatory phenotype of CD1c+ DC2 and CD163+ DC3 with increased CD56 and CD15 in dendritic cells and monocytes. Moreover, the blood of RA patients contains HLA-DR+ cells with shared dendritic cell and granulocytic features. These phenotypic characterizations of RA patients implicate CD1c+ DC2 and CD163+ DC3 in the systemic autoimmune disease rheumatoid arthritis and suggest that increased HLA-DR+ phenotypes with shared granulocytic and dendritic cell features can contribute to RA, potentially by providing enhanced co-stimulatory presentation of self-antigen(s) to CD4+ T lymphocytes.
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spelling doaj-art-72d6f0d23bff480b8cb4f81fbfed2bd72025-08-20T03:28:14ZengFrontiers Media S.A.Frontiers in Immunology1664-32242025-07-011610.3389/fimmu.2025.15966091596609Spectral cytometry of rheumatoid arthritis patients implicates myeloid dendritic cells and granular HLA-DR+CD15+CD16+ cells in pro-inflammatory antigen presentationChristian Geier0Haani Qudsi1Estelle Khairallah2Jihad Ben Gabr3Robert Winchester4Andras Perl5Division of Rheumatology and Clinical Immunology, Department of Medicine, State University of New York Upstate Medical University, Syracuse, NY, United StatesNorton College of Medicine, State University of New York Upstate Medical University, Syracuse, NY, United StatesNorton College of Medicine, State University of New York Upstate Medical University, Syracuse, NY, United StatesDivision of Rheumatology and Clinical Immunology, Department of Medicine, State University of New York Upstate Medical University, Syracuse, NY, United StatesNorton College of Medicine, State University of New York Upstate Medical University, Syracuse, NY, United StatesDivision of Rheumatology and Clinical Immunology, Department of Medicine, State University of New York Upstate Medical University, Syracuse, NY, United StatesIntroductionRheumatoid arthritis (RA) is a systemic autoimmune disease that leads to inflammation of synovial joints and other organs. Many RA patients “share” a common peptide sequence within the HLA-DR (MHC II) molecule expressed on antigen-presenting cells (APC), suggesting that HLA-DR+ cells are important in RA inflammation. We use HLA-DR positivity to comprehensively immunophenotype APC by spectral cytometry.MethodsWe measured mean fluorescence intensities (MFI) of HLA-DR and molecules associated with dendritic cells (CD141, CD1c, CD163, CD11c, CD123, and CD303), monocytes (CD14 and CD16), granulocytic markers (CD15 and CCR3), co-stimulatory molecules (CD86 and CD275), and chemokine receptors (CCR2, CCR3, and CCR7) from RA patients and healthy donors by spectral flow cytometry.ResultsDC2 (CD1c+) showed higher CD86, CD275 (ICOS-L), CD56, and CCR7 in RA (all p < 0.05). CD56 was also increased in (CD163+) DC3 (p = 0.0453). CD15 was increased throughout RA dendritic cell subsets and classical and intermediate monocytes (all p < 0.01). Except for B cells, HLA-DR was not different in RA. A distinct HLA-DR+CD15+CD16+ population appeared in RA (p = 0.0004), which contributed a mean of 1.3% (± SD 2.85%) to the overall HLA-DR+ APC compartment. This HLA-DR+CD15+CD16+ subset was positive for CD83, CD275, and, like plasmacytoid pDC, CD303+. However, in contrast to pDC, it formed distinct t-SNE clusters and differed from reference pDC (CD123+CD303+) by much less CD123 (p < 0.01). The HLA-DR+CD15+CD16+ phenotype correlated with clinical markers of systemic inflammation (p < 0.01).DiscussionIn conclusion, dendritic cell and monocyte alterations in RA include an increased co-stimulatory phenotype of CD1c+ DC2 and CD163+ DC3 with increased CD56 and CD15 in dendritic cells and monocytes. Moreover, the blood of RA patients contains HLA-DR+ cells with shared dendritic cell and granulocytic features. These phenotypic characterizations of RA patients implicate CD1c+ DC2 and CD163+ DC3 in the systemic autoimmune disease rheumatoid arthritis and suggest that increased HLA-DR+ phenotypes with shared granulocytic and dendritic cell features can contribute to RA, potentially by providing enhanced co-stimulatory presentation of self-antigen(s) to CD4+ T lymphocytes.https://www.frontiersin.org/articles/10.3389/fimmu.2025.1596609/fullrheumatoid arthritisCD1c+ dendritic cellsintermediate monocytesmyeloid cellslow-density granulocytesantigen-presenting cell
spellingShingle Christian Geier
Haani Qudsi
Estelle Khairallah
Jihad Ben Gabr
Robert Winchester
Andras Perl
Spectral cytometry of rheumatoid arthritis patients implicates myeloid dendritic cells and granular HLA-DR+CD15+CD16+ cells in pro-inflammatory antigen presentation
Frontiers in Immunology
rheumatoid arthritis
CD1c+ dendritic cells
intermediate monocytes
myeloid cells
low-density granulocytes
antigen-presenting cell
title Spectral cytometry of rheumatoid arthritis patients implicates myeloid dendritic cells and granular HLA-DR+CD15+CD16+ cells in pro-inflammatory antigen presentation
title_full Spectral cytometry of rheumatoid arthritis patients implicates myeloid dendritic cells and granular HLA-DR+CD15+CD16+ cells in pro-inflammatory antigen presentation
title_fullStr Spectral cytometry of rheumatoid arthritis patients implicates myeloid dendritic cells and granular HLA-DR+CD15+CD16+ cells in pro-inflammatory antigen presentation
title_full_unstemmed Spectral cytometry of rheumatoid arthritis patients implicates myeloid dendritic cells and granular HLA-DR+CD15+CD16+ cells in pro-inflammatory antigen presentation
title_short Spectral cytometry of rheumatoid arthritis patients implicates myeloid dendritic cells and granular HLA-DR+CD15+CD16+ cells in pro-inflammatory antigen presentation
title_sort spectral cytometry of rheumatoid arthritis patients implicates myeloid dendritic cells and granular hla dr cd15 cd16 cells in pro inflammatory antigen presentation
topic rheumatoid arthritis
CD1c+ dendritic cells
intermediate monocytes
myeloid cells
low-density granulocytes
antigen-presenting cell
url https://www.frontiersin.org/articles/10.3389/fimmu.2025.1596609/full
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