Stanley J R, Koulu L, Klaus-Kovtun V, Steinberg M S
J Immunol. 1986 Feb 15;136(4):1227-30.
Recent studies have demonstrated that antibodies from about half of patients with pemphigus foliaceus (PF) bind to a 160 kd polypeptide ("PF antigen") in sodium dodecyl sulfate (SDS) extracts of normal human epidermis. Desmoglein (DG) I, a glycoprotein enriched in desmosomal cores, is approximately the same m.w. as PF antigen. To demonstrate that PF autoantibodies bind to DG I, we used a monoclonal IgG antibody (MmDGI-1) that was raised against bovine muzzle desmosomal cores, and that specifically binds DG I. Double immunofluorescence labeling was performed on the same section of normal human skin with PF antibodies, detected by fluorescein-conjugated goat anti-human IgG, and MmDGI-1, detected by rhodamine-conjugated goat anti-mouse IgG. The pattern of reactivity with both antibodies was identical. Immunoblotting studies on proteins extracted from normal human epidermis and separated by SDS-polyacrylamide gel electrophoresis demonstrated that PF antibodies and MmDGI-1 bound co-migrating polypeptide bands of approximate m.w. 160,000. To confirm that these were identical polypeptides, we performed immunoblots of these epidermal extracts that were separated by two-dimensional gel electrophoreses (isoelectric focusing followed by SDS-PAGE). PF antibodies and MmDGI-1 bound identical spots with pI approximately 5.4 to 5.7 and m.w. approximately 160,000. These studies demonstrate that autoantibodies from certain patients with PF, a disorder of cell adhesion, bind to DG I, a desmosomal core glycoprotein.
最近的研究表明,约一半落叶型天疱疮(PF)患者的抗体可与正常人表皮十二烷基硫酸钠(SDS)提取物中的一种160kd多肽(“PF抗原”)结合。桥粒芯糖蛋白(DG)I是一种富含于桥粒芯的糖蛋白,其分子量与PF抗原大致相同。为了证明PF自身抗体与DG I结合,我们使用了一种针对牛口鼻部桥粒芯产生的单克隆IgG抗体(MmDGI-1),该抗体可特异性结合DG I。用异硫氰酸荧光素偶联的山羊抗人IgG检测PF抗体,用罗丹明偶联的山羊抗小鼠IgG检测MmDGI-1,对正常人皮肤的同一切片进行双重免疫荧光标记。两种抗体的反应模式相同。对从正常人表皮提取并经SDS-聚丙烯酰胺凝胶电泳分离的蛋白质进行免疫印迹研究表明,PF抗体和MmDGI-1结合了分子量约为160,000的共迁移多肽条带。为了证实这些是相同的多肽,我们对经二维凝胶电泳(等电聚焦后进行SDS-PAGE)分离的这些表皮提取物进行了免疫印迹。PF抗体和MmDGI-1结合了相同的斑点,其等电点约为5.4至5.7,分子量约为160,000。这些研究表明,来自某些PF患者(一种细胞黏附障碍疾病)的自身抗体可与桥粒芯糖蛋白DG I结合。