Identification of a primary biliary cirrhosis associated protein as lysosome-associated membrane protein-2
Identification of a primary biliary cirrhosis associated protein as lysosome-associated membrane protein-2
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原发性胆汁性肝硬化相关蛋白的鉴定为溶酶体相关膜蛋白-2
DOI：
10.1016/j.jprot.2013.08.019
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发表时间：
2013-10-08
影响因子：
3.3
通讯作者：
Han, Ying
中科院分区：
文献类型：
--
作者：
Wang, Lu;Wang, Jingbo;Han, Ying
原发性胆汁性肝硬化（PBC）是一种病因不明的慢性胆汁淤积性肝病，其发病机制可能与肝胆转运异常有关。在这项研究中，我们的目的是分离和鉴定与PBC相关的新分子。以PBC患者肝细胞小管膜囊泡（CMV）为免疫原，筛选出特异性识别正常肝细胞亚顶端区的单克隆抗体1F 9（mAb 1F 9）。免疫组化结果显示，PBC肝组织中mAb 1F 9抗原（mAb 1F 9-Ag）的表达明显高于正常肝组织、肝硬化及胆汁淤积组。有趣的是，mAb 1F 9-Ag的表达增加与PBC的严重程度相关，熊去氧胆酸治疗可显著改善mAb 1F 9-Ag的恢复。46%的PBC中存在mAb 1F 9-Ag的再分布。分离纯化mAb 1F 9-Ag，经质谱分析，确定其为溶酶体相关膜蛋白2（lysosome-associated membrane protein 2，LAMP-2）。进一步的研究表明，mAb 1F 9识别LAMP-2免疫沉淀物，反之，mAb 1F 9与重组LAMP-2反应。mAb 1F 9和LAMP-2抗体显示相似的染色模式，并显示相似的亚细胞定位。总之，mAb 1F 9-Ag的身份是LAMP-2，这表明LAMP-2可能有助于PBC的分化，并预测PBC患者的不良结局。mAb 1F 9识别的抗原可能有助于原发性胆汁性肝硬化（PEG）的分化，并预测PEG患者的不良结局。通过抗原鉴定，我们确认mAb 1F 9-Ag的身份为溶酶体相关膜蛋白2（LAMP-2）。该手稿的临床相关性得到了很好的评价，因为标记物很少见，通常不能成功用于PBC的诊断和治疗。(C)2013爱思唯尔有限公司版权所有。
Primary biliary cirrhosis (PBC) is a chronic cholestatic liver disease of unknown etiology and abnormality of hepatobiliary transport might contribute to its pathogenesis. In this study, we aimed to isolate and identify new molecules associated with PBC. With hepatocyte canalicular membrane vesicles (CMVs) of PBC patients as immunogens, we screened the monoclonal antibody 1F9 (mAb1F9), whose antigen dominantly recognized the subapical domains in hepatocytes in normal livers. Immunohistochemistry revealed that the expression of mAb1F9 antigen (mAb1F9-Ag) significantly increased in PBC livers compared with control groups including normal livers, cirrhosis or cholestasis other than PBC. Interestingly, the augmented expression of mAb1F9-Ag was correlated with the severity of PBC, and ursodeoxycholic acid treatment may significantly improve the recovery of mAb1F9-Ag. In addition, redistribution of mAb1F9-Ag was found in 46% of PBC. mAb1F9-Ag was isolated and analyzed with mass spectrometry, which indicated lysosome-associated membrane protein 2 (LAMP-2) as the candidate. Further studies showed that mAb1F9 recognized LAMP-2 immunoprecipitates and vice verse, mAb1F9 reacted with recombinant LAMP-2. mAb1F9 and LAMP-2 antibody exhibited similar staining pattern and displayed similar subcellular localization. Together, the identity of mAb1F9-Ag is LAMP-2, suggesting that LAMP-2 may assist in the differentiation of PBC and predict a poor outcome in patients with PBC.Biological significanceThis manuscript describes the expression of a specific antibody, named mAb1F9. The antigen recognized by mAb1F9 may assist in the differentiation of primary biliary cirrhosis (PEG) and predict a poor outcome in patients with PEG. Through antigen identification, we confirm the identity of mAb1F9-Ag as lysosome-associated membrane protein 2 (LAMP-2). The clinical relevance of the manuscript is well regarded since markers are rare and usually not successful for PBC diagnosis and treatment. (C) 2013 Elsevier B.V. All rights reserved.