Identification of novel autoantigens via mass spectroscopy-based antibody-mediated identification of autoantigens (MS-AMIDA) using immune thrombocytopenic purpura (ITP) as a model disease
Identification of novel autoantigens via mass spectroscopy-based antibody-mediated identification of autoantigens (MS-AMIDA) using immune thrombocytopenic purpura (ITP) as a model disease
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DOI：
10.1016/j.jprot.2017.01.012
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发表时间：
2017-03-22
影响因子：
3.3
通讯作者：
Salama, Abdulgabar
中科院分区：
文献类型：
--
作者：
Kamhieh-Milz, Julian;Sterzer, Viktor;Salama, Abdulgabar
免疫性血小板减少性紫癜（ITP）是特征最明确的自身免疫性疾病之一。针对血小板抗原的自身抗体 (AAB) 被认为是 ITP 的诊断标志，但仅在 50% 的患者中可检测到。我们设计并应用了一种新型蛋白质组学方法，称为基于质谱的抗体介导的自身抗原鉴定（MS-AMIDA），用于血小板抗原。患者被分为具有经典 AAB 的患者 [ITP(+)] 和不具有 AAB5 的患者 [ITP(-)]。总共，在 ITP(+) 中发现了 181 个潜在 AAG，在 ITP(-) 中发现了 135 个 AAG，在两次 MS-AMIDA 运行中重复发现了 34 个和 23 个 AAG。从对照中减去标识符后，ITP(+) 中保留了 57 个 AAG，ITP(+) 中保留了 29 个 AAG，1TP(+) 和 ITP(-) 患者中常见有 16 个 AAG。无标记定量 (LFQ) 揭示了 15 种潜在的 AAG，它们在 ITP 中的定量更强。对己糖激酶 1 (HK1)、El 丙酮酸脱氢酶 (El-PDH)、凝血因子 XIII、细丝蛋白 A (FLNA)、非肌肉肌球蛋白 9 进行斑点印迹验证。发现 11 名患者具有抗 HK1 AAB5，一名患者具有抗 El-PDH AAB，两名患者具有抗 FLNA AAB。大多数抗原是细胞内来源的，与肌动蛋白细胞骨架和程序性细胞死亡的调节显着相关。总之，使用 MS-AMIDA 鉴定了用于 ITP 的新型 AAG。 (C) 2017 Elsevier B.V. 保留所有权利。
Immune thrombocytopenic purpura (ITP) is one of the best characterized autoimmune diseases. Autoantibodies (AABs) against platelet antigens are considered as the diagnostic hallmark of ITP, but are detectable in only 50% of patients. We designed and applied a novel proteomic approach termed Mass Spectroscopy-based Antibody-Mediated Identification of Autoantigens (MS-AMIDA) for platelet antigens. Patients were separated into patients with classical AABs [ITP(+)] and patients without AAB5 [ITP(-)]. Altogether, 181 potential AAGs were found in ITP(+) and 135 AAGs in ITP(-), with 34 and 23 AAGs reproducibly found in two runs of MS-AMIDA. After subtracting identifiers from the controls, 57 AAGs in ITP(+) and 29 AAGs in ITP(+) remained, with 16 AAGs commonly found in 1TP(+) and ITP(-) patients. Label-free quantification (LFQ) revealed 15 potential AAGs that are quantitatively stronger in ITP. Dot blot validation was performed on hexokinase 1 (HK1), El pyruvate dehydrogenase (El-PDH), coagulation factor XIII, filamin A (FLNA), non-muscle myosin 9. Eleven patients were found to have anti-HK1 AAB5, one patient had anti-El-PDH AABs, and two patients had anti-FLNA AABs. Most antigens were of intracellular origin with significant association with actin-cytoslceleton and regulation of programmed cell death. In conclusion, novel AAGs for ITP were identified using MS-AMIDA. (C) 2017 Elsevier B.V. All rights reserved.