Servettaz Amélie, Tamby Mathieu C, Guilpain Philippe, Reinbolt Joseph, Garcia de la Penã-Lefebvre Paloma, Allanore Yannick, Kahan André, Meyer Olivier, Guillevin Loïc, Mouthon Luc
Université Paris--Descartes, faculté de Médecine, UPRES-EA 1833, Laboratoire d'Immunologie, Paris, France.
Clin Immunol. 2006 Aug;120(2):212-9. doi: 10.1016/j.clim.2006.02.006. Epub 2006 Mar 31.
By using a quantitative immunoblotting technique on protein extracts of human macrovascular and microvascular endothelial cells, we have analyzed the self-reactive repertoires of IgG from 20 patients with limited cutaneous SSc, 40 patients with diffuse SSc and 60 age- and sex-matched healthy controls. Serum IgG from 15/20 patients with limited cutaneous SSc and anti-centromere antibodies bound to at least one of the two 75- and 85-kDa protein bands in the different endothelial cell extracts, whereas IgG from healthy controls or patients with diffuse SSc did not. N-terminal sequencing of the 75- and 85-kDa bands identified CENP-B as the sole antigen in both bands. Moreover, IgG from all of the SSc patients who recognized the 75- and/or 85-kDa bands bound to a full-length recombinant CENP-B protein as assessed by ELISA, whereas IgG from other SSc patients did not. The main target of anti-endothelial cell antibodies in patients with limited cutaneous SSc is the nuclear and ubiquitous protein CENP-B.
通过对人大血管和微血管内皮细胞的蛋白质提取物使用定量免疫印迹技术，我们分析了20例局限性皮肤系统性硬化症患者、40例弥漫性皮肤系统性硬化症患者以及60例年龄和性别匹配的健康对照者的IgG自身反应谱。15/20例局限性皮肤系统性硬化症且抗着丝点抗体阳性患者的血清IgG与不同内皮细胞提取物中的两条75 kDa和85 kDa蛋白条带中的至少一条结合，而健康对照者或弥漫性皮肤系统性硬化症患者的IgG则未出现这种情况。对75 kDa和85 kDa条带进行N端测序后确定CENP - B是两条条带中的唯一抗原。此外，通过酶联免疫吸附测定（ELISA）评估，所有识别75 kDa和/或85 kDa条带的系统性硬化症患者的IgG均与全长重组CENP - B蛋白结合，而其他系统性硬化症患者的IgG则未结合。局限性皮肤系统性硬化症患者抗内皮细胞抗体的主要靶标是细胞核内普遍存在的蛋白CENP - B。
