Novel autoimmune hepatitis-specific autoantigens identified using protein microarray technology.

Novel autoimmune hepatitis-specific autoantigens identified using protein microarray technology.
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DOI:
10.1021/pr900131e
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发表时间:
2010-01
影响因子:
4.4
通讯作者:
Wu L
Wu L
中科院分区:
生物学2区
文献类型:
--
作者:
Song Q;Liu G;Hu S;Zhang Y;Tao Y;Han Y;Zeng H;Huang W;Li F;Chen P;Zhu J;Hu C;Zhang S;Li Y;Zhu H;Wu L

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自身免疫性肝炎(AIH)是一种慢性肝脏炎症坏死性疾病,其病因尚不清楚。使用免疫血清学方法检测非器官特异性和肝脏相关自身抗体已广泛用于诊断和预后。然而,明确且准确地检测疾病需要识别和表征疾病特异性自身抗原。在本研究中,我们分析了 AIH 患者与其他肝病患者的自身抗原库,识别并验证了三种新型且高度特异性的 AIH 生物标志物。在第一阶段,我们制造了包含 5,011 个非冗余蛋白质的人类蛋白质芯片,并用它以相对较少的血清收集量快速识别 11 种候选自身抗原。在第二阶段,我们制造了一种 AIH 特异性蛋白质芯片,并获得了 44 名 AIH 患者、50 名健康对照者和另外 184 名患有乙型肝炎、丙型肝炎、系统性红斑狼疮、原发性干燥综合征、类风湿性关节炎或原发性胆汁性肝硬化患者的血清样本的自身免疫原性特征。利用这种两阶段方法,我们鉴定了三种新抗原:RPS20、Alba-like 和 dUTPase,作为高度 AIH 特异性的生物标志物,其敏感性分别为 47.5% (RPS20)、45.5%(Alba-like)和 22.7%(dUTPase)。这些潜在的生物标志物在双盲设计中通过额外的 AIH 样本得到了进一步验证。最后,我们证明这些新的生物标志物可以很容易地应用于基于 ELISA 的检测,用于临床诊断/预后。
Autoimmune hepatitis (AIH) is a chronic necroinflammatory disease of the liver with a poorly understood etiology. Detection of non-organ-specific and liver-related autoantibodies using immunoserological approaches has been widely used for diagnosis and prognosis. However, unambiguous and accurate detection of the disease requires the identification and characterization of disease-specific autoantigens. In the present study, we have profiled the autoantigen repertoire of patients with AIH versus those with other liver diseases, identifying and validating three novel and highly specific biomarkers for AIH. In Phase I we fabricated a human protein chip of 5,011 non-redundant proteins and used it to quickly identify 11 candidate autoantigens with relative small serum collection. In Phase II we fabricated an AIH-specific protein chip and obtained autoimmunogenic profiles of serum samples from 44 AIH patients, 50 healthy controls, and 184 additional patients suffering from hepatitis B, hepatitis C, systemic lupus erythematosus, primary Sjögren’s syndrome, rheumatoid arthritis, or primary biliary cirrhosis. Using this two-phase approach, we identified three new antigens, RPS20, Alba-like, and dUTPase, as highly AIH-specific biomarkers, with sensitivities of 47.5% (RPS20), 45.5% (Alba-like), and 22.7% (dUTPase). These potential biomarkers were further validated with additional AIH samples in a double-blind design. Finally, we demonstrated that these new biomarkers could be readily applied to ELISA-based assays for use in clinical diagnosis/prognosis.
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发表时间: 2002-03-01
期刊: NATURE MEDICINE
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期刊: ARTHRITIS RESEARCH
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