Koulu L, Kusumi A, Steinberg M S, Klaus-Kovtun V, Stanley J R
J Exp Med. 1984 Nov 1;160(5):1509-18. doi: 10.1084/jem.160.5.1509.
Pemphigus foliaceus (PF) is a human autoimmune disease in which antibodies are directed against the cell surface of epidermal cells with resultant blister formation. The histopathology of these blisters indicates that cells have detached from each other, and electron microscopy of early blisters shows diminished numbers, to complete loss, of desmosomes as well as abnormalities of the tonofilament-desmosome complex. In this study we demonstrate that autoantibodies from certain PF patients bind to a desmosomal core glycoprotein called desmoglein (DG) I. Proteins in extracts of normal human epidermis were separated by sodium dodecyl sulfate (SDS) polyacrylamide gel electrophoresis (PAGE), then transferred to nitrocellulose or 2-aminophenylthioether paper for immunoperoxidase staining. Results of these immunoblots indicated that sera from 6 of 13 PF patients specifically and intensely stained an approximately 160,000 mol wt polypeptide, "PF antigen". Such staining was not seen with normal human sera or sera from patients with pemphigus vulgaris or bullous pemphigoid, two autoimmune blistering skin diseases that are clinically, histologically, and immunochemically distinct from PF. However, rabbit antiserum directed against DGI, that was isolated from bovine muzzle desmosomes, stained a polypeptide band which co-migrated with PF antigen. Furthermore, when proteins from extracts of normal human epidermis were electrophoresed in two dimensions (isoelectric focusing, then SDS-PAGE) before transfer to nitrocellulose for immunoperoxidase staining, PF antibodies and antibodies to DGI stained identical spots. Finally, PF sera as well as PF IgG that was affinity purified with PF antigen from normal human epidermis, both selectively bound to DGI extracted from bovine muzzle desmosomes. These studies demonstrate that the human autoantibodies from certain patients with PF, a disease of epidermal cell adhesion, are directed against a desmosomal core protein.
落叶型天疱疮(PF)是一种人类自身免疫性疾病,其中抗体针对表皮细胞的细胞表面,导致水疱形成。这些水疱的组织病理学表明细胞彼此分离,早期水疱的电子显微镜检查显示桥粒数量减少直至完全丧失,以及张力丝 - 桥粒复合体异常。在本研究中,我们证明来自某些PF患者的自身抗体与一种称为桥粒芯糖蛋白(DG)I的桥粒核心糖蛋白结合。正常人表皮提取物中的蛋白质通过十二烷基硫酸钠(SDS)聚丙烯酰胺凝胶电泳(PAGE)分离,然后转移到硝酸纤维素膜或2 - 氨基苯硫醚纸上进行免疫过氧化物酶染色。这些免疫印迹结果表明,13例PF患者中有6例的血清特异性且强烈地染色了一种约160,000摩尔质量的多肽,即“PF抗原”。正常人血清或寻常型天疱疮或大疱性类天疱疮患者的血清未出现这种染色,这两种自身免疫性水疱性皮肤病在临床、组织学和免疫化学上与PF不同。然而,从牛口鼻部桥粒中分离出的针对DG I的兔抗血清染色了一条与PF抗原共迁移的多肽带。此外,当正常人表皮提取物中的蛋白质在二维电泳(等电聚焦,然后SDS - PAGE)后转移到硝酸纤维素膜上进行免疫过氧化物酶染色时,PF抗体和针对DG I的抗体染色相同的斑点。最后,PF血清以及用正常人表皮中的PF抗原亲和纯化的PF IgG,都选择性地结合从牛口鼻部桥粒中提取的DG I。这些研究表明,来自某些PF患者(一种表皮细胞黏附疾病)的人类自身抗体针对一种桥粒核心蛋白。