Labib R S, Camargo S, Futamura S, Martins C R, Rock B, Anhalt G J, Diaz L A
Department of Dermatology, Johns Hopkins University, Baltimore, Maryland.
J Invest Dermatol. 1989 Aug;93(2):272-9. doi: 10.1111/1523-1747.ep12277591.
In both the endemic and sporadic forms of pemphigus foliaceus (PF), antiepidermal autoantibodies against desmoglein I are present. Desmoglein I is a highly insoluble 160-kD transmembrane glycoprotein of the desmosomal core. The detailed immunochemical characterization of the epitope(s) recognized by the PF autoantibodies is hampered by its large molecular weight and the insolubility of desmoglein I in nondenaturing buffers. This study was designed to identify alternative methods that could yield soluble immunoreactive PF antigen (Ag) from normal human epidermis. The presence of PF Ag in human epidermis and in its soluble or insoluble fractions was monitored by indirect immunofluorescence, immunoadsorption of PF sera, and immunoprecipitation of radiolabeled fractions. The PF Ag from trypsin-resistant, radiolabeled cell envelope preparations was cleaved by papain and immunoprecipitated by PF sera. A 50-kD peptide, isoelectric at pH 5.5-5.8, was immunoprecipitated by sera from all patients with endemic PF (n = 15) or idiopathic PF (n = 4), and by two of four pemphigus vulgaris sera, but by no control sera (n = 7). This study shows that a significant fraction of the PF Ag is insoluble, trypsin-resistant, and is associated with the cornified cell envelope fraction, but an Ag fragment can be obtained in a small molecular weight, soluble, and immunoreactive form by papain digestion. This 50-kD papain fragment is more amenable to detailed chemical and immunologic characterization than the native molecule.
在落叶型天疱疮(PF)的地方性和散发性两种形式中,均存在针对桥粒芯糖蛋白1的抗表皮自身抗体。桥粒芯糖蛋白1是桥粒核心的一种高度不溶性的160-kD跨膜糖蛋白。PF自身抗体识别的表位的详细免疫化学特征因桥粒芯糖蛋白1的大分子质量以及其在非变性缓冲液中的不溶性而受到阻碍。本研究旨在确定能够从正常人表皮产生可溶性免疫反应性PF抗原(Ag)的替代方法。通过间接免疫荧光、PF血清的免疫吸附以及放射性标记组分的免疫沉淀来监测人表皮及其可溶性或不溶性组分中PF Ag的存在。来自抗胰蛋白酶、放射性标记的细胞包膜制剂的PF Ag被木瓜蛋白酶切割并被PF血清免疫沉淀。一种50-kD的肽,其等电点在pH 5.5 - 5.8,被所有地方性PF患者(n = 15)或特发性PF患者(n = 4)的血清以及4例寻常型天疱疮血清中的2例血清免疫沉淀,但未被对照血清(n = 7)免疫沉淀。本研究表明,相当一部分PF Ag是不溶性的、抗胰蛋白酶的,并且与角化细胞包膜组分相关,但通过木瓜蛋白酶消化可以获得一种小分子质量、可溶性且具有免疫反应性的Ag片段。这种50-kD的木瓜蛋白酶片段比天然分子更易于进行详细的化学和免疫学特征分析。