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For targeted therapies it is important to understand site-specific cellular adaptations. Juvenile idiopathic arthritis (JIA) is characterized by autoimmune joint inflammation, with frequent inadequate treatment responses. To comprehensively assess the inflammatory immune landscape, we designed a 37-parameter spectral flow cytometry panel delineating mononuclear cells from JIA synovial fluid (SF) of autoimmune inflamed joints, compared to JIA and healthy control blood. Synovial monocytes and NK cells (CD56bright) lack Fc-receptor CD16, suggesting antibody-mediated targeting may be ineffective. B cells and DCs, both in small frequencies in SF, undergo maturation with high 4-1BB, CD71, CD39 expression, supporting T-cell activation. SF effector and regulatory T cells were highly active with newly described co-receptor combinations that may alter function, and suggestion of metabolic reprogramming via CD71, TNFR2, and PD-1. Most SF effector phenotypes, as well as an identified CD4-Foxp3+ T-cell population, were restricted to the inflamed joint, yet specific SF-predominant CD4+ Foxp3+ Treg subpopulations were increased in blood of active but not inactive JIA, suggesting possible recirculation and loss of immunoregulation at distal sites. This first comprehensive dataset of the site-specific inflammatory landscape at protein level will inform functional studies and the development of targeted therapeutics to restore immunoregulatory balance and achieve remission in JIA.<\/jats:p>","DOI":"10.1093\/cei\/uxae071","type":"journal-article","created":{"date-parts":[[2024,8,2]],"date-time":"2024-08-02T09:45:09Z","timestamp":1722591909000},"page":"221-241","source":"Crossref","is-referenced-by-count":12,"title":["The immune landscape of the inflamed joint defined by spectral flow cytometry"],"prefix":"10.1093","volume":"218","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-3996-8740","authenticated-orcid":false,"given":"Meryl H","family":"Attrill","sequence":"first","affiliation":[{"name":"Institute of Immunity and Transplantation, Division of Infection and Immunity, University College London , London ,","place":["UK"]},{"name":"UCL Great Ormond Street Institute of Child Health, Infection, Immunity, and Inflammation Research 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