Aiba S, Masuko T, Yagita H, Hashimoto Y, Tagami H
Department of Dermatology, Tohoku University School of Medicine, Sendai, Japan.
Acta Derm Venereol. 1987;67(5):399-405.
MoAb AHS-7 labels the basement membrane of the skin. In this report, we have determined the precise localization and molecular nature of the antigen(s). After epidermal-dermal separation by suction blistering, the antigen(s) were detected on the epidermal side. Immunoelectron microscopy revealed the reaction product observed as a discontinuous line along the plasma membrane of the basal keratinocytes, which corresponded to the distribution of the hemidesmosomes. We extracted the proteins from the normal human epidermis separated from the dermis by suction blistering. After being separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, the proteins were electrophoretically transferred to nitrocellulose sheets, and the antigen(s) were identified by immunoperoxidase staining with MoAb AHS-7. We detected 4 protein bands of molecular weights of 82 kd, 104 kd, 120-135 kd, and 140-160 kd when unreduced, in contrast to only two bands of 41 kd and 52 kd when reduced. These data suggest that the antigen(s) recognized by MoAb AHS-7 defines a unique antigen, different from bullous pemphigoid antigen on the hemidesmosome zone.
单克隆抗体AHS - 7标记皮肤的基底膜。在本报告中,我们已确定了抗原的精确定位和分子性质。通过水疱抽吸实现表皮 - 真皮分离后,在表皮侧检测到了抗原。免疫电子显微镜显示反应产物表现为沿着基底角质形成细胞质膜的不连续线,这与半桥粒的分布相对应。我们从通过水疱抽吸与真皮分离的正常人表皮中提取蛋白质。经十二烷基硫酸钠 - 聚丙烯酰胺凝胶电泳分离后,将蛋白质电泳转移至硝酸纤维素膜上,并用单克隆抗体AHS - 7通过免疫过氧化物酶染色鉴定抗原。未还原时,我们检测到分子量分别为82 kd、104 kd、120 - 135 kd和140 - 160 kd的4条蛋白带,而还原时仅检测到41 kd和52 kd的两条带。这些数据表明,单克隆抗体AHS - 7识别的抗原定义了一种独特的抗原,不同于半桥粒区的大疱性类天疱疮抗原。