Sakly Nabil, Mirshahi Pejman, Soria Jeannette, Ghedira Ibtissem, Mirshahi Massoud
Faculty of Pharmacy, Rue Avicenne, Monastir 5000, Tunisia.
Ann N Y Acad Sci. 2005 Jun;1050:201-9. doi: 10.1196/annals.1313.021.
Cyto-ELISA has been widely used to investigate anti-endothelial cell antibodies (AECAs); however, because various types of endothelial cells have been used, the results among studies differ. The aim of our study was to analyze and compare the results when determining AECAs in patients with connective tissue disease (CTD). We did so using a cyto-ELISA with different cells as antigenic substrates: two different endothelial cells, one microvascular (HMEC-1) and one from human bone marrow (HBMEC), and one epithelial cell line from breast adenocarcinoma as negative controls (MDA-MB-231). In this trial, we performed a retrospective study in 60 patients with CTD [46 with systemic lupus erythematosus, 8 with Sjögren's syndrome, and 6 with systemic sclerosis] and 32 healthy volunteers. Using cyto-ELISA, the antibody against a cell was considered positive when the optical density (OD) obtained was higher than the mean OD obtained in the control group + 2 standard deviations (upper normal range). Patients were classified into three groups according to the OD obtained with the different cell lines: group 1: patients without any antibody; group 2: patients with specific AECAs; and group 3: patients with nonspecific AECAs. According to this classification, we found that 43.3% of patients with CTD have specific AECAs, and 28.3% have nonspecific antibodies. Our study delineates the heterogeneity of AECAs in patients with CTD. The use of HBMEC in cyto-ELISA may increase the sensitivity of the test, and the use of nonendothelial cells as negative controls may improve its specificity.
细胞酶联免疫吸附测定（Cyto-ELISA）已被广泛用于研究抗内皮细胞抗体（AECAs）；然而，由于使用了多种类型的内皮细胞，各研究结果存在差异。我们研究的目的是分析和比较在结缔组织病（CTD）患者中检测AECAs时的结果。我们使用以不同细胞作为抗原底物的细胞酶联免疫吸附测定来进行此项研究：两种不同的内皮细胞，一种微血管内皮细胞（HMEC-1）和一种人骨髓来源的内皮细胞（HBMEC），以及一种来自乳腺腺癌的上皮细胞系作为阴性对照（MDA-MB-231）。在该试验中，我们对60例CTD患者[46例系统性红斑狼疮患者、8例干燥综合征患者和6例系统性硬化症患者]和32名健康志愿者进行了一项回顾性研究。使用细胞酶联免疫吸附测定时，当获得的光密度（OD）高于对照组获得的平均OD + 2个标准差（正常上限范围）时，针对某一细胞的抗体被视为阳性。根据用不同细胞系获得的OD将患者分为三组：第1组：无任何抗体的患者；第2组：具有特异性AECAs的患者；第3组：具有非特异性AECAs的患者。根据这种分类，我们发现43.3%的CTD患者具有特异性AECAs，28.3%的患者具有非特异性抗体。我们的研究描述了CTD患者中AECAs的异质性。在细胞酶联免疫吸附测定中使用HBMEC可能会提高检测的敏感性，使用非内皮细胞作为阴性对照可能会提高其特异性。
