Cooley J E, Briggaman R A, Cronce D J, Banes A J, O'Keefe E J
Department of Dermatology, University of North Carolina, Chapel Hill 27599, USA.
J Invest Dermatol. 1996 Dec;107(6):877-81. doi: 10.1111/1523-1747.ep12331167.
The blisters in the inherited disorder, Hailey-Hailey disease, may be caused by defective epidermal junctional complexes. We evaluated these structural complexes in vivo and in vitro. We induced a vesicular lesion in the apparently normal skin of a patient with Hailey-Hailey disease and studied a biopsy of this lesion by transmission electron microscopy. To determine whether acantholysis was related to a defect in the number or assembly of intercellular junctions, we cultured Hailey-Hailey disease keratinocytes in medium containing 0.1 mM Ca2+ and increased the [Ca2+] to 1.1 mM in order to induce assembly of cell-cell junctions. Keratinocytes were examined by double immunofluorescence with antibodies to the desmosome protein, desmoplakin, and the adherens junction protein, vinculin, at intervals after the increase in [Ca2+]. Characteristic Hailey-Hailey disease histopathology was observed by electron microscopy of the patient's skin after trauma, but we found no splitting of desmosomes. Based on the location, intensity, and rate of change of immunofluorescent staining, Hailey-Hailey and normal keratinocytes did not differ in their ability to assemble desmosomes and adherens junctions. Furthermore, we observed no significant morphologic differences between normal and Hailey-Hailey keratinocytes cultured in low and high [Ca2+]-containing media; Hailey-Hailey cells contained abundant normal-appearing desmosomes in 1.1 mM [Ca2+]. Since Hailey-Hailey disease keratinocytes can assemble normal-appearing adherens junctions and desmosomes in vitro, the functional defect may not lie in assembly of cell-cell adhering junctions, or additional perturbation may be required to expose the defect.
遗传性疾病海利-海利病（Hailey-Hailey disease）中的水疱可能由有缺陷的表皮连接复合体引起。我们在体内和体外对这些结构复合体进行了评估。我们在一名海利-海利病患者看似正常的皮肤上诱导出一个水疱性病变，并通过透射电子显微镜对该病变的活检组织进行研究。为了确定棘层松解是否与细胞间连接数量或组装缺陷有关，我们在含有0.1 mM Ca2+的培养基中培养海利-海利病角质形成细胞，并将[Ca2+]增加到1.1 mM以诱导细胞间连接的组装。在[Ca2+]增加后的不同时间间隔，用针对桥粒蛋白桥粒斑蛋白（desmoplakin）和黏附连接蛋白纽蛋白（vinculin）的抗体进行双重免疫荧光检查角质形成细胞。对患者皮肤创伤后的电子显微镜观察发现了典型的海利-海利病组织病理学特征，但我们未发现桥粒的分裂。根据免疫荧光染色的位置、强度和变化速率，海利-海利病角质形成细胞和正常角质形成细胞在组装桥粒和黏附连接的能力上没有差异。此外，我们观察到在含低[Ca2+]和高[Ca2+]培养基中培养的正常角质形成细胞和海利-海利病角质形成细胞之间没有明显的形态学差异；在1.1 mM [Ca2+]条件下海利-海利病细胞含有大量外观正常的桥粒。由于海利-海利病角质形成细胞在体外能够组装外观正常的黏附连接和桥粒，功能缺陷可能不在于细胞间黏附连接的组装，或者可能需要额外的干扰才能暴露该缺陷。
