Identification of candidate endothelial cell autoantigens in systemic lupus erythematosus using a molecular cloning strategy: a role for ribosomal P protein P0 as an endothelial cell autoantigen
Identification of candidate endothelial cell autoantigens in systemic lupus erythematosus using a molecular cloning strategy: a role for ribosomal P protein P0 as an endothelial cell autoantigen
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DOI：
10.1093/rheumatology/39.10.1114
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发表时间：
2000-10-01
期刊：
影响因子：
5.5
通讯作者：
Murphy, JJ
中科院分区：
文献类型：
--
作者：
Frampton, G;Moriya, S;Murphy, JJ
目标。目的：利用分子克隆技术研究系统性红斑狼疮（SLE）患者抗内皮细胞抗体（AECA）识别抗原的多样性和性质。采用ELISA法检测15例SLE患者血清AECA水平，采用Western blot法检测2例临床活动性SLE患者自身抗原靶点的多样性。用这两例患者的血清免疫筛选人脐静脉内皮细胞cDNA表达文库，以确定其自身抗原靶点。采用抗核糖体P肽抗体ELISA法评价1例患者血清中抗核糖体P蛋白抗体的临床意义。5例活动性疾病和肾炎患者的AECA水平明显高于5例临床非活动性疾病患者。两名临床活跃患者的血清可识别不同的自身抗原谱。候选自身抗原包括：(1)内皮细胞特异性纤溶酶原激活物抑制剂；(2)经典狼疮抗原，即核糖体P蛋白P0；(3)以前从未被描述为SLE推定的自身抗原的蛋白质，包括核糖体蛋白L6、延伸因子1 α、腺苷酸环化酶相关蛋白、DNA复制许可因子、profilin II和新蛋白HEAPLA 1和HEAPLA 2（人内皮相关推定狼疮自身抗原1和2）。在一项纵向研究中，在一名患者中，针对核糖体P蛋白P0的抗体占主导地位，这些抗体的水平与总AECA水平、抗dna抗体滴度、总体临床评分和肾脏疾病相关。一组候选的SLE内皮自身抗原，包括先前描述的自身抗原和新的靶点，已经通过分子克隆策略确定。这种新的分子方法也可以应用于其他自身免疫性血管疾病的自身抗原鉴定。
Objective. To attempt to characterize the diversity and nature of antigens recognized by anti-endothelial cell antibodies (AECA) in patients with systemic lupus erythematosus (SLE) using a molecular cloning strategy.Methods. AECA in sera of 15 SLE patients were measured by ELISA and Western blot analysis was used to examine the diversity of autoantigen targets in two clinically active patients. A human umbilical vein endothelial cell cDNA expression library was immunoscreened with sera from these two patients to identify their autoantigen targets. An anti-ribosomal P peptide antibody ELISA was used to assess the clinical significance of anti-ribosomal P protein antibodies in the sera of one patient.Results. Significantly higher AECA levels were found in five patients with active disease and nephritis than in five patients with clinically inactive disease. Sera from two clinically active patients were found to recognize distinct spectra of autoantigens. The candidate autoantigens that were identified included (1) endothelial cell-specific plasminogen activator inhibitor, (2) the classical lupus antigen, i.e. ribosomal P protein P0; and (3) proteins never before described as putative autoantigens in SLE, including ribosomal protein L6, elongation factor 1 alpha, adenyl cyclase-associated protein, DNA replication licensing factor, profilin II and the novel proteins HEAPLA 1 and HEAPLA 2 (human endothelial associated putative lupus autoantigens 1 and 2). In one patient, antibodies against ribosomal P protein P0 were predominant and levels of these antibodies correlated with total AECA levels, anti-DNA antibody titres, overall clinical score and renal disease in a longitudinal study.Conclusions. A panel of candidate endothelial autoantigens in SLE, which includes previously described autoantigens and novel targets, has been identified by a molecular cloning strategy. This novel molecular approach could also be applied to the identification of autoantigens in other autoimmune vascular diseases.