外周血中中性粒细胞胞外陷阱（NETs）形成增加于少关节性、多关节性幼年特发性关节炎及腱鞘炎相关关节炎：作为诊断和疾病活动性的生物标志物

Increased neutrophil extracellular trap formation in oligoarticular, polyarticular juvenile idiopathic arthritis and enthesitis-related arthritis: biomarkers for diagnosis and disease activity

Frontiers in Immunology

影响因子:5.9

分区:医学2区 / 免疫学2区

发表日期:2024

作者: Hongxia Tang, Yucheng Zhong, Yali Wu, Yanmei Huang, Yi Liu, Jing Chen, Ting Xi, Yini Wen, Ting He, Shanshan Yang, Fan Liu, Runji Xiong, Runming Jin

DOI: 10.3389/fimmu.2024.1436193

摘要

中性粒细胞胞外陷阱（NETs）在启动和维持炎症过程中起重要作用。然而，NETs在不同亚型幼年特发性关节炎（JIA）中的作用尚少有研究。因此，本研究旨在探讨JIA来源的中性粒细胞释放NETs的能力，TNF-α（肿瘤坏死因子-α）抑制剂对NETs形成的影响（包括体外和体内），以及评估NETs衍生产物与临床和免疫相关参数的关联。通过体外刺激和抑制研究评估中性粒细胞释放NETs的能力以及阿达木单抗对NET形成的影响。检测血浆中的NET衍生产物以评估体内NET形成的发生率。此外，采用流式细胞术和Western blot检测中性粒细胞中的NET相关信号组分。与健康对照（HC）相比，少关节型JIA、多关节型JIA以及腱鞘炎相关关节炎患者的中性粒细胞更易于自发性和响应TNF-α或PMA产生NETs。在外周血中，JIA患者存在过度的NET形成，血浆中NET衍生产物（游离DNA和MPO-DNA复合物）水平升高，能准确区分JIA患者与HC，并正相关于疾病活动度。多元线性回归分析显示，红细胞沉降率和TNF-α水平为独立变量，且与游离DNA浓度呈正相关。值得注意的是，TNF-α抑制剂在体外和体内均能有效阻止NET的形成。此外，JIA来源的中性粒细胞中与NET相关的激酶磷酸化水平显著升高。研究表明，NET在JIA的发病机制中可能具有致病作用，并可能参与TNF-α介导的炎症过程。血浆中NET衍生产物具有潜在的诊断和疾病监测价值。进一步的研究揭示了JIA患者中NET形成的分子机制，为NET靶向治疗提供理论基础。

Abstract

Neutrophil extracellular traps (NETs) are important factors in initiating and perpetuating inflammation. However, the role of NETs in different subtypes of juvenile idiopathic arthritis (JIA) has been rarely studied. Therefore, we aimed to explore the ability of JIA-derived neutrophils to release NETs and the effect of TNF-α (tumor necrosis factor-alpha) inhibitors on NET formation both in vitro and in vivo, and evaluate the associations of NET-derived products with clinical and immune-related parameters.The ability of neutrophils to release NETs and the effect of adalimumab on NET formation was assessed via in vitro stimulation and inhibition studies. Plasma NET-derived products were detected to assess the incidence of NET formation in vivo. Furthermore, flow cytometry and western blotting were used to detect NET-associated signaling components in neutrophils.Compared to those derived from HCs, neutrophils derived from patients with oligoarticular-JIA, polyarticular-JIA and enthesitis-related arthritis were more prone to generate NETs spontaneously and in response to TNF-α or PMA in vitro. Excessive NET formation existed in peripheral circulation of JIA patients, and elevated plasma levels of NET-derived products (cell-free DNA and MPO-DNA complexes) could accurately distinguish JIA patients from HCs and were positively correlated with disease activity. Multiple linear regression analysis showed that erythrocyte sedimentation rate and TNF-α levels were independent variables and were positively correlated with cell-free DNA concentration. Notably, TNF-α inhibitors could effectively prevent NET formation both in vitro and in vivo. Moreover, the phosphorylation levels of NET-associated kinases in JIA-derived neutrophils were markedly increased.Our data suggest that NETs might play pathogenic roles and may be involved in TNF-α-mediated inflammation in JIA. Circulating NET-derived products possess potential diagnostic and disease monitoring value. Furthermore, the preliminary results related to the molecular mechanisms of NET formation in JIA patients provide a theoretical basis for NET-targeted therapy.