Department of Rheumatology, Peking Union Medical College Hospital, Chinese Academy of Medical Science & Peking Union Medical College, National Clinical Research Center for Dermatologic and Immunologic Diseases, State Key Laboratory of Complex Severe and Rare Diseases, #1 Shuai-Fu-Yuan, Dongcheng District, Beijing, 100730, China.
Clinical Biobank, Department of Medical Research Center, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China.
Arthritis Res Ther. 2022 Jul 19;24(1):170. doi: 10.1186/s13075-022-02860-4.
Neutrophils and aberrant NETosis have been implicated in the pathogenesis of diverse autoimmune diseases; however, their roles in primary Sjögren's syndrome (pSS) remain unclear. We aimed to reveal the potential roles of neutrophils and neutrophil extracellular traps (NETs) in pSS.
pSS patients were enrolled and NETosis markers were measured in plasma and labial glands using ELISA and immunofluorescence. The gene signatures of neutrophils were assessed by RNA-Seq and RT-PCR. Reactive oxygen species (ROS), mitochondrial ROS (MitoSOX) production, and JC-1 were measured by flow cytometry.
NETosis markers including cell-free DNA (cf-DNA) and myeloperoxidase (MPO) in plasma and labial glands from pSS patients were significantly higher than healthy controls (HCs) and were associated with disease activity. RNA sequencing and RT-qPCR revealed activated type I IFN signaling pathway and higher expression of genes related to type I interferon in pSS neutrophils. Further stimulating with IFN-α 2a in vitro significantly induced ROS production and JC-1 monomer percentage in pSS neutrophils.
Our data suggest the involvement of neutrophils and enhanced NETosis in pSS patients. Further mechanism study in vitro revealed that type I IFN activation in pSS neutrophils led to mitochondrial damage and related ROS production which finally result in the generation of NETs.
中性粒细胞和异常细胞 NETosis 已被牵连在多种自身免疫性疾病的发病机制中；然而，它们在原发性干燥综合征 (pSS) 中的作用仍不清楚。我们旨在揭示中性粒细胞和中性粒细胞胞外诱捕网 (NETs) 在 pSS 中的潜在作用。
招募 pSS 患者，并使用 ELISA 和免疫荧光法测量血浆和唇腺中的 NETosis 标志物。通过 RNA-Seq 和 RT-PCR 评估中性粒细胞的基因特征。通过流式细胞术测量活性氧 (ROS)、线粒体 ROS (MitoSOX) 的产生和 JC-1。
pSS 患者血浆和唇腺中的 NETosis 标志物，包括细胞游离 DNA (cf-DNA) 和髓过氧化物酶 (MPO)，明显高于健康对照 (HC)，并与疾病活动度相关。RNA 测序和 RT-qPCR 显示 pSS 中性粒细胞中激活的 I 型干扰素信号通路和与 I 型干扰素相关的基因表达更高。体外进一步用 IFN-α 2a 刺激显着增加了 pSS 中性粒细胞中的 ROS 产生和 JC-1 单体百分比。
我们的数据表明中性粒细胞和增强的 NETosis 参与了 pSS 患者。体外进一步的机制研究表明，pSS 中性粒细胞中 I 型 IFN 的激活导致线粒体损伤和相关的 ROS 产生，最终导致 NETs 的产生。
