Boulinguez S, Bedane C, Cadilhac H, Grolleau-Rochiccioli P, Caux F, Oksman F, Bonafé J L
Service de Dermatologie, Hôptial Rangueil, Toulouse.
Ann Dermatol Venereol. 1995;122(6-7):417-21.
Pemphigus vulgaris (PV) and pemphigus foliaceus (PF) are autoimmune blistering diseases characterized by loss of cell-cell adhesion and by autoantibodies directed against epidermal cadherins. The ultrastructural localization of PV antigen remains controversial, whereas the location of PF antigen seems to be established. The use of different techniques could explain these various data. To investigate this matter, indirect immunoelectron microscopy (IEM) and Western blot analysis on bovine tongue epithelium were used.
Serum samples from patients with PF(3), PV (4) and control samples from healthy patients (2) were analysed in this study. The inclusion criteria were based upon characteristic clinical features, level of epidermal cleavage on histological preparations and presence of circulating anti-epithelial cell surface antibodies. Indirect IME was performed on normal human skin. Peroxidase labelling was used. Serum samples were also analysed by western immunoblotting on bovine tongue epithelium.
Indirect IEM examination of PV sera showed immune deposits located both on desmosomal and extra-desmosomal areas, whereas in PF, IgG deposits were strictly localized on desmosomal structures. By Western blot analysis, PV sera recognized a 130 kDa polypeptide and PF sera a 150 kDa polypeptide.
Indirect IEM on normal human skin using peroxidase labelling was used because of the best antigenic conservation obtained. Our results suggest that PV antigen could exist both on desmosomal junctions and adherens junctions, whereas PF antigen (desmoglein I) is restricted to desmosome.
寻常型天疱疮(PV)和落叶型天疱疮(PF)是自身免疫性水疱病,其特征为细胞间黏附丧失以及针对表皮钙黏蛋白的自身抗体。PV抗原的超微结构定位仍存在争议,而PF抗原的定位似乎已明确。使用不同技术可能解释这些不同的数据。为研究此问题,我们对牛舌上皮进行了间接免疫电子显微镜(IEM)和蛋白质印迹分析。
本研究分析了来自PF患者(3例)、PV患者(4例)的血清样本以及健康患者的对照样本(2例)。纳入标准基于特征性临床特征、组织学标本上表皮松解水平以及循环抗上皮细胞表面抗体的存在情况。在正常人皮肤上进行间接IME,采用过氧化物酶标记。血清样本也通过对牛舌上皮进行免疫印迹分析。
PV血清的间接IEM检查显示免疫沉积物位于桥粒和非桥粒区域,而在PF中,IgG沉积物严格定位于桥粒结构上。通过蛋白质印迹分析,PV血清识别出一条130 kDa的多肽,PF血清识别出一条150 kDa的多肽。
由于获得了最佳的抗原保存效果,因此使用了过氧化物酶标记的正常人皮肤间接IEM。我们的结果表明,PV抗原可能存在于桥粒连接和黏着连接上,而PF抗原(桥粒芯糖蛋白I)仅限于桥粒。