Objectives: A number of immune populations have been implicated in psoriatic arthritis (PsA) pathogenesis. This study used mass cytometry (CyTOF) combined with transcriptomic analysis to generate a high-dimensional dataset of matched PsA synovial fluid (SF) and blood leucocytes, with the aim of identifying cytokine production ex vivo in unstimulated lymphoid and myeloid cells. Methods: Fresh SF and paired blood were either fixed or incubated with protein transport inhibitors for 6 h. Samples were stained with two CyTOF panels: a phenotyping panel and an intracellular panel, including antibodies to both T cell and myeloid cell secreted proteins. Transcriptomic analysis by gene array of key expanded cell populations and single-cell RNA-seq, and ELISA and LEGENDplex analysis of PsA SF were also performed. Results: We observed marked changes in the myeloid compartment of PsA SF relative to blood, with expansion of intermediate monocytes, macrophages and dendritic cell populations. Classical monocytes, intermediate monocytes and macrophages spontaneously produced significant levels of the proinflammatory mediators osteopontin and CCL2 in the absence of any in vitro stimulation. By contrast minimal spontaneous cytokine production by T cells was detected. Gene expression analysis showed the genes for osteopontin and CCL2 to be amongst those most highly upregulated by PsA monocytes/macrophages; and both proteins were elevated in PsA SF. Conclusions: Using multiomic analyses we have generated a comprehensive cellular map of PsA SF and blood to reveal key expanded myeloid proinflammatory modules in PsA of potential pathogenic and therapeutic importance.

中文翻译：

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体外大量细胞计数分析揭示了牛皮癣关节炎滑液中的深刻的髓样促炎信号

目的：许多免疫人群与银屑病关节炎（PsA）的发病机制有关。这项研究使用大规模细胞计数（CyTOF）结合转录组学分析来生成匹配的PsA滑液（SF）和血白细胞的高维数据集，目的是鉴定未受刺激的淋巴和髓样细胞离体的细胞因子产生。方法：将新鲜的SF和成对的血液固定或与蛋白质转运抑制剂一起孵育6小时。样品用两个CyTOF板染色：一个表型板和一个细胞内板，包括针对T细胞和髓样细胞分泌蛋白的抗体。还通过关键扩展细胞群和单细胞RNA-seq的基因阵列进行了转录组学分析，并对PsA SF进行了ELISA和LEGENDplex分析。 结果：我们观察到相对于血液，PsA SF的髓样区室发生了显着变化，中间单核细胞，巨噬细胞和树突状细胞群体均发生了扩张。在没有任何体外刺激的情况下，经典单核细胞，中间单核细胞和巨噬细胞会自发产生大量促炎性介质骨桥蛋白和CCL2。相反，检测到由T细胞产生的最小自发细胞因子产生。基因表达分析表明，骨桥蛋白和CCL2基因属于PsA单核细胞/巨噬细胞上调程度最高的基因之一。并且两种蛋白质在PsA SF中均升高。 结论： 使用多组学分析，我们已经生成了PsA SF和血液的全面细胞图，以揭示PsA中潜在的潜在致病和治疗重要性的关键扩展髓样促炎模块。