A novel autoantibody against fibronectin leucine-rich transmembrane protein 2 expressed on the endothelial cell surface identified by retroviral vector system in systemic lupus erythematosus.
A novel autoantibody against fibronectin leucine-rich transmembrane protein 2 expressed on the endothelial cell surface identified by retroviral vector system in systemic lupus erythematosus.
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一种针对纤连蛋白亮氨酸亮氨酸的跨膜蛋白2的新型自身抗体，该蛋白2在全身性红斑狼疮的逆转录病毒载体系统鉴定出的内皮细胞表面上表达。
DOI：
10.1186/ar3897
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发表时间：
2012-07-02
影响因子：
4.9
通讯作者：
Harigae H
中科院分区：
文献类型：
--
作者：
Shirai T;Fujii H;Ono M;Nakamura K;Watanabe R;Tajima Y;Takasawa N;Ishii T;Harigae H
抗内皮细胞抗体(AECA)被认为在胶原疾病的血管炎症中起关键作用，但大多数是针对细胞内的分子，而不是表达在细胞表面。为了阐明AECs的致病作用，我们构建了一个逆转录病毒载体系统，用于鉴定表达于内皮细胞表面的自身抗原。以人脐静脉内皮细胞(HUVECs)为材料，用流式细胞仪检测胶原病患者血清中AECA活性。将人脐静脉内皮细胞(HUVECs)的cDNA文库逆转录病毒导入大鼠骨髓瘤细胞系，用流式细胞仪对AECA阳性克隆进行筛选。对细胞进行DNA分析，鉴定自身抗原，并对自身抗体的临床特征和功能意义进行评价。从1例系统性红斑狼疮(SLE)患者血清免疫球蛋白G(Ig G)中分离到两个不同的AECA阳性克隆。两个克隆均与富含亮氨酸的纤维连接蛋白跨膜蛋白2(Flrt2)的基因序列一致。人脐静脉内皮细胞表达Flrt2，表达的AECA免疫球蛋白可与转染人Flt2的细胞特异性结合。SLE患者抗Flrt2抗体活性占AECA的21.4%。此外，抗Flrt2抗体对表达flrt2的细胞具有补体依赖性细胞毒作用。我们利用逆转录病毒载体系统鉴定了膜蛋白Flrt2是SLE患者AECAs的一种新的自身抗原。抗Flrt2抗体有可能在约10%的SLE患者中诱导内皮细胞的直接细胞毒作用，可能成为一种新的干预分子靶点。识别AECAs的这种细胞表面靶点可能揭示胶原疾病中血管损伤的综合机制。
Anti-endothelial cell antibodies (AECAs) are thought to be critical for vasculitides in collagen diseases, but most were directed against molecules localized within the cell and not expressed on the cell surface. To clarify the pathogenic roles of AECAs, we constructed a retroviral vector system for identification of autoantigens expressed on the endothelial cell surface. AECA activity in sera from patients with collagen diseases was measured with flow cytometry by using human umbilical vein endothelial cells (HUVECs). A cDNA library of HUVECs was retrovirally transfected into a rat myeloma cell line, from which AECA-positive clones were sorted with flow cytometry. cDNA of the cells was analyzed to identify an autoantigen, and then the clinical characteristics and the functional significance of the autoantibody were evaluated. Two distinct AECA-positive clones were isolated by using serum immunoglobulin G (IgG) from a patient with systemic lupus erythematosus (SLE). Both clones were identical to cDNA of fibronectin leucine-rich transmembrane protein 2 (FLRT2). HUVECs expressed FLRT2 and the prototype AECA IgG bound specifically to FLRT2-transfected cells. Anti-FLRT2 antibody activity accounted for 21.4% of AECAs in SLE. Furthermore, anti-FLRT2 antibody induced complement-dependent cytotoxicity against FLRT2-expressing cells. We identified the membrane protein FLRT2 as a novel autoantigen of AECAs in SLE patients by using the retroviral vector system. Anti-FLRT2 antibody has the potential to induce direct endothelial cell cytotoxicity in about 10% of SLE patients and could be a novel molecular target for intervention. Identification of such a cell-surface target for AECAs may reveal a comprehensive mechanism of vascular injury in collagen diseases.
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影响因子：
--
作者：
MASI, AT;HUNDER, GG;ZVAIFLER, NJ
通讯作者：
ZVAIFLER, NJ
影响因子：
4.6
作者：
Bordron, A;Révélen, R;Youinou, P
通讯作者：
Youinou, P
影响因子：
4.4
作者：
Lacy, SE;Bönnemann, CG;Kunkel, LM
通讯作者：
Kunkel, LM
影响因子：
--
作者：
Dieudé, M;Senécal, JL;Raymond, Y
通讯作者：
Raymond, Y
影响因子：
13.6
作者：
Alard, Jean-Eric;Dueymes, Maryvonne;Jamin, Christophe
通讯作者：
Jamin, Christophe