Identification of L-plastin autoantibody in plasma of patients with non-Hodgkin's lymphoma using a proteomics-based analysis
Identification of L-plastin autoantibody in plasma of patients with non-Hodgkin's lymphoma using a proteomics-based analysis
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DOI：
10.1258/acb.2007.006230
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发表时间：
2008-01-01
影响因子：
2.2
通讯作者：
Matsuura, Nariaki
中科院分区：
文献类型：
--
作者：
Ueda, Kazuhito;Nakanishi, Toyofumi;Matsuura, Nariaki
背景：非霍奇金淋巴瘤（NHL）和霍奇金淋巴瘤（HL）等恶性淋巴瘤（ML）的诊断主要依靠形态学检查和基因分析。只有少量的血清/血浆生物标志物，如乳酸脱氢酶和可溶性白细胞介素-2受体α，可用于诊断ML。分类多种多样，因此使用流式细胞术或淋巴结活检检查了细胞表面标志物。然而，区分NHL和HL这两种疾病是很困难的。方法：为了鉴定患者血浆中血液学恶性肿瘤相关的自身免疫反应性（自身抗体），采用了一种基于蛋白质组学的电泳/质谱分析方法。通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳和Western blotting分析分离伯基特淋巴瘤细胞系（Raji）的溶解蛋白，其中血液系统恶性肿瘤患者的血浆检测一抗，然后用辣根过氧化物酶偶联的抗igg抗体可视化。结果：采用基质辅助激光解吸电离/飞行时间质谱法和串联质谱法检测了NHL患者血浆中L-plastin和α -烯醇化酶两种能与抗体发生反应的蛋白。NHL患者抗l -活素自身抗体的检出率显著高于NHL患者：0.84 (21/25)；HL组0.00 (0/4)；自身免疫性疾病0.38 (5/13)；白血病为0.20 (2/10)；健康对照组为0.13（1/8）。而抗-烯醇化酶抗体对NHL无特异性。结论：我们首次在NHL患者血浆中检测到l -活蛋白自身抗体，提示该自身抗体可作为NHL新的诊断生物标志物。
Background: The diagnosis of malignant lymphoma (ML) such as non-Hodgkin's lymphoma (NHL) and Hodgkin's lymphoma (HL) was mainly performed by morphological examination and gene analysis. There are only a few serum/plasma biomarkers such as lactate dehydrogenase and soluble interleukin-2 receptor alpha to diagnose ML. The classifications are various, and therefore the cell surface markers using flow cytometry or lymph node biopsy have been examined. It is difficult, however, to distinguish the two diseases, NHL and HL, from each other.Methods: In order to identify the haematological malignancy-associated autoimmunoreactivity (autoantibodies) in patients' plasma, a novel proteomics-based approach using electrophoresis/mass spectrometry was applied. Solubilized proteins from a Burkitt's lymphoma cell line (Raji) were separated by sodium dodecyl sulphate-polyacrylamide gel electrophoresis and Western blotting analysis, in which the plasma of individual patients with haematological malignancies was tested for primary antibodies, followed by visualization with anti-IgG antibody conjugated with horseradish peroxidase.Results: Two proteins, L-plastin and alpha-enolase, capable of reacting with the antibodies in plasma of patients with NHL, were detected using matrix-assisted laser desorption ionization/time-of-flight mass spectrometry and tandem mass spectrometry. The rates of the detections of an anti L-plastin autoantibody were significantly higher: 0.84 (21/25) in patients with NHL; 0.00 (0/4) in HL; 0.38 (5/13) in autoimmune diseases; 0.20 (2/10) in leukaemia; and 0.13 (1/8) in healthy controls. In contrast, those of anti alpha-enolase antibody were not specific to NHL.Conclusions: We first identified autoantibody against L-plastin in plasma of patients with NHL, suggesting that the autoantibody can be a new diagnostic biomarker for NHL.