通过人类蛋白质微阵列鉴定自身免疫性肝炎患者体内的抗DOK2抗体。
Identification of anti-DOK2 antibodies in patients with autoimmune hepatitis via a human protein microarray.
作者信息
Abe Kazumichi, Wada Jun, Hayashi Manabu, Sugaya Tatsuro, Abe Naoto, Takahata Yosuke, Fujita Masashi, Takahashi Atsushi, Kanno Yukiko, Kuroda Masahito, Saito Keietsu, Nozawa Yoshihiro, Migita Kiyoshi, Yatsuhashi Hiroshi, Ohira Hiromasa
机构信息
Department of Gastroenterology, Fukushima Medical University School of Medicine, Fukushima, Japan.
Department of Internal Medicine, Hanawa Kosei Hospital, Higashishirakawa, Japan.
出版信息
Hepatol Commun. 2025 Sep 22;9(10). doi: 10.1097/HC9.0000000000000807. eCollection 2025 Oct 1.
BACKGROUND
Autoantibodies such as antinuclear antibodies (ANAs) and anti-smooth muscle antibodies (ASMAs), which are detected in many cases of autoimmune hepatitis (AIH), are not disease-specific; thus, they have limited value in disease diagnosis. In this study, we aimed to identify disease-specific autoantibodies related to AIH diagnosis or pathogenesis.
METHODS
We analyzed human protein microarrays (containing more than 16,000 human proteins, including membrane, cytoplasmic, and nuclear proteins, synthesized in a wheat germ cell-free system using cDNA). To identify AIH-specific autoantibodies, we performed a cross-sectional study of AIH patients (n=67) and matched healthy controls (HCs) (n=25) via ELISA. An independent cohort of 123 AIH patients and 75 HCs was analyzed for validation. All patient data were collected before corticosteroid therapy. We also evaluated the associations between candidate protein antigen expression and clinical findings in AIH patients. Candidate protein antigen expression in liver tissues was confirmed by immunohistochemistry.
RESULTS
According to comprehensive human protein microarray analysis, docking protein 2 (DOK2) was selected from 6 antigens upregulated in AIH patients. The mean anti-DOK2 level was significantly greater in AIH patients (36.1-37.4 AU) than in those with other liver diseases (2.6-9.2) or HCs (1.0-4.5 AU) in the exploration and validation cohorts (p value/AUROC, vs. HCs, <0.0001/0.87-0.90). Serum anti-DOK2 levels were positively correlated with serum IgG levels and liver histological activity. The serum IgG level was an independent predictor of increased anti-DOK2 antibody levels in patients with AIH. Immunostaining revealed that DOK2-positive cells were present in the hepatic lobules and portal tracts of patients with AIH.
CONCLUSIONS
We provide the first evidence that serum anti-DOK2 antibody levels are associated with the diagnosis and disease activity of AIH patients.
背景
自身抗体，如在许多自身免疫性肝炎（AIH）病例中检测到的抗核抗体（ANA）和抗平滑肌抗体（ASMA），并非疾病特异性的；因此，它们在疾病诊断中的价值有限。在本研究中，我们旨在鉴定与AIH诊断或发病机制相关的疾病特异性自身抗体。
方法
我们分析了人类蛋白质微阵列（包含超过16000种人类蛋白质，包括膜蛋白、细胞质蛋白和核蛋白，在无细胞小麦胚芽系统中使用cDNA合成）。为了鉴定AIH特异性自身抗体，我们通过酶联免疫吸附测定（ELISA）对AIH患者（n = 67）和匹配的健康对照者（HCs）（n = 25）进行了横断面研究。对123例AIH患者和75例HCs的独立队列进行分析以作验证。所有患者数据均在皮质类固醇治疗前收集。我们还评估了AIH患者中候选蛋白抗原表达与临床发现之间的关联。通过免疫组织化学证实肝组织中候选蛋白抗原的表达。
结果
根据全面的人类蛋白质微阵列分析，从AIH患者中上调的6种抗原中选择了对接蛋白2（DOK2）。在探索和验证队列中，AIH患者的平均抗DOK2水平（36.1 - 37.4 AU）显著高于其他肝病患者（2.6 - 9.2）或HCs（1.0 - 4.5 AU）（p值/AUROC，与HCs相比，<0.0001/0.87 - 0.90）。血清抗DOK2水平与血清IgG水平和肝脏组织学活性呈正相关。血清IgG水平是AIH患者抗DOK2抗体水平升高的独立预测因子。免疫染色显示，AIH患者的肝小叶和门管区存在DOK2阳性细胞。
结论
我们提供了首个证据，表明血清抗DOK2抗体水平与AIH患者的诊断和疾病活动相关。
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