Won Taejoon, Naik Pooja, Wood Megan K, Wang Hong, Talor Monica V, Shi Jing, Bracamonte-Baran William, Thomas Mekha A, Jaime Camille M, Jo Wonyoung, Ray Shreyanshu, Foss Catherine A, Andrade Felipe, Čiháková Daniela, Darrah Erika
Johns Hopkins University School of Medicine, Baltimore, Maryland, and College of Veterinary Medicine, University of Illinois Urbana-Champaign, Urbana, Illinois.
Johns Hopkins University School of Medicine, Baltimore, Maryland.
Arthritis Rheumatol. 2025 Apr 2. doi: 10.1002/art.43168.
Autoantibodies targeting peptidylarginine deiminase 4 (PAD4), an enzyme involved in protein citrullination, are found in a subset of patients with rheumatoid arthritis (RA) with severe joint disease. However, the mechanisms by which anti-PAD4 antibodies participate in disease pathogenesis are incompletely defined.
We investigated the role of anti-PAD4 monoclonal antibodies derived from patients with RA using a collagen-induced arthritis (CIA) mouse model and human monocyte in vitro cultures. The cellular targets of anti-PAD4 antibodies were identified using mouse knee joint cells and human peripheral blood mononuclear cells. In addition, PAD4 gene and protein expression was assessed using human fibroblast-like synoviocyte in vitro cultures and a single-cell RNA sequencing data set obtained from patients with RA.
We show that anti-PAD4 antibody treatment augmented disease severity in the CIA mouse model, with increased joint damage, myeloid cell infiltration, and synovial fibroblast activation. Arthritic mice administered with anti-PAD4 antibodies had an increased proportion of interleukin-17A (IL-17A), tumor necrosis factor α (TNFα), and interferon-γ (IFNγ)-producing T cells. Anti-PAD4 antibodies preferentially bound monocytes in both humans and mice, eliciting proinflammatory chemokine production by human monocytes in vitro. T cell cytokines enhanced by anti-PAD4 antibodies in the CIA model (ie, IL-17A, TNFα, and IFNγ) synergized to induce a proinflammatory phenotype in human fibroblast-like synoviocytes.
Our findings suggest a model in which anti-PAD4 antibody binding to monocytes triggers an inflammatory cascade that promotes immune cell recruitment to the joint and T cell activation, culminating in synovial fibroblast activation and the development of more severe arthritis.
在患有严重关节疾病的类风湿关节炎(RA)患者亚组中发现了靶向肽基精氨酸脱氨酶4(PAD4)的自身抗体,该酶参与蛋白质瓜氨酸化。然而,抗PAD4抗体参与疾病发病机制的方式尚未完全明确。
我们使用胶原诱导性关节炎(CIA)小鼠模型和人单核细胞体外培养,研究了源自RA患者的抗PAD4单克隆抗体的作用。使用小鼠膝关节细胞和人外周血单个核细胞鉴定抗PAD4抗体的细胞靶点。此外,使用人成纤维细胞样滑膜细胞体外培养和从RA患者获得的单细胞RNA测序数据集评估PAD4基因和蛋白表达。
我们发现抗PAD4抗体治疗加重了CIA小鼠模型的疾病严重程度,关节损伤增加、髓样细胞浸润和滑膜成纤维细胞活化。给予抗PAD4抗体的关节炎小鼠产生白细胞介素-17A(IL-17A)、肿瘤坏死因子α(TNFα)和干扰素-γ(IFNγ)的T细胞比例增加。抗PAD4抗体在人和小鼠中均优先结合单核细胞,在体外诱导人单核细胞产生促炎趋化因子。在CIA模型中,抗PAD4抗体增强的T细胞细胞因子(即IL-17A、TNFα和IFNγ)协同作用,在人成纤维细胞样滑膜细胞中诱导促炎表型。
我们的研究结果提示了一种模型,即抗PAD4抗体与单核细胞结合触发炎症级联反应,促进免疫细胞募集到关节并激活T细胞,最终导致滑膜成纤维细胞活化和更严重关节炎的发展。