Hintner H, Stingl G, Schuler G, Fritsch P, Stanley J, Katz S, Wolff K
J Invest Dermatol. 1981 Feb;76(2):113-8. doi: 10.1111/1523-1747.ep12525447.
The classification of mechanobullous diseases often depends on the electron microscopic distinction of intradermal (dermolytic), junctional and intraepidermal sites of cleavage. Electron microscopy is tedious and time consuming. In this report we describe a different approach to the determination of the cleavage plane by using a method which recognizes subtle differences in the localization of antigenic structures relative to the cleavage plane. Cryostat sections of lesional and extralesional skin of 3 patients with dermolytic epidermolysis bullosa, 3 with epidermolytic epidermolysis bullosa and 8 with junctional epidermolysis bullosa were examined by immunofluorescence, with specific antisera against type IV collagen (localized within the basal lamina); against laminin (noncollagenous protein, localized in the lamina lucida); and with bullous pemphigoid antibodies (directed against the bullous pemphigoid antigen localized in the lamina lucida). All specimens were also examined by electron microscopy. In dermolytic epidermolysis bullosa (where cleft formation occurs intradermally) type IV collagen, laminin and the bullous pemphigoid antigen were consistently found in the roof of the blister, whereas in junctional epidermolysis bullosa (where the cleft occurs in the lamina lucida) type IV collagen and laminin were found on the floor of the blister whereas bullous pemphigoid antigen was present mainly on the roof, but focally also on the floor, of the blister. In epidermolytic epidermolysis bullosa (where the cleft is intraepidermal) all antigens were localized below the cleavage plane. In all cases electron microscopy confirmed the level of cleft formation predicted from the immunofluorescence mapping of the antigenic sites. The described method equals electron microscopy in accuracy but it is more rapid and simpler to perform.
机械性大疱性疾病的分类通常取决于真皮内（皮肤松解性）、交界性和表皮内分裂部位的电子显微镜鉴别。电子显微镜检查繁琐且耗时。在本报告中，我们描述了一种不同的方法来确定分裂平面，即使用一种能够识别抗原结构相对于分裂平面定位的细微差异的方法。对3例皮肤松解性大疱性表皮松解症患者、3例表皮松解性大疱性表皮松解症患者和8例交界性大疱性表皮松解症患者的皮损及皮损外皮肤的冰冻切片进行免疫荧光检查，使用针对IV型胶原（位于基底层内）、层粘连蛋白（非胶原性蛋白，位于透明层）的特异性抗血清以及大疱性类天疱疮抗体（针对位于透明层的大疱性类天疱疮抗原）。所有标本也进行了电子显微镜检查。在皮肤松解性大疱性表皮松解症（裂隙形成于真皮内）中，IV型胶原、层粘连蛋白和大疱性类天疱疮抗原始终位于水疱顶部，而在交界性大疱性表皮松解症（裂隙发生于透明层）中，IV型胶原和层粘连蛋白位于水疱底部，而大疱性类天疱疮抗原主要存在于水疱顶部，但也有局灶性地存在于水疱底部。在表皮松解性大疱性表皮松解症（裂隙位于表皮内）中，所有抗原均位于分裂平面下方。在所有病例中，电子显微镜证实了从抗原位点的免疫荧光定位预测的裂隙形成水平。所描述的方法在准确性上与电子显微镜相当，但执行起来更快且更简单。
