Ceilley E, Watanabe N, Shapiro D, Verrando P, Bauer E A, Burgeson R, Briggaman R A, Woodley D T
Department of Dermatology, Stanford University School of Medicine, CA.
J Dermatol Sci. 1993 Apr;5(2):97-103. doi: 10.1016/0923-1811(93)90077-3.
A variety of methods were used to fracture the dermal-epidermal junction (DEJ) of human skin. These included warm and hot phosphate buffered saline, trypsin, cold 1 M salt, potassium bromide and proteolytic digestion with dispase. The localization and sensitivity of basement membrane components (bullous pemphigoid antigen, BM 600/nicein, epiligrin, kalinin, laminin, collagens IV and VII (EBA antigen) and linkin) were determined after the DEJ was fractured by each method. We found that the basement membrane zone proteins, BM 600/nicein, epiligrin and kalinin remained with the dermal side of the DEJ fractured through the lamina lucida by cold salt, phosphate buffered saline and potassium bromide. BM 600/nicein, epiligrin and kalinin were not detected after treatment with trypsin. In contrast, laminin, another glycoprotein in the lamina lucida, was insensitive to all of the procedures, but co-localized to the dermal side of DEJ-fractured skin. We also found that separation of the DEJ with brief exposure of skin to 56 degrees C provided a useful substrate for testing the autoantibodies in the sera of patients with epidermolysis bullosa acquisita (EBA). Heat-separated skin can be prepared in a significantly shorter period of time than salt-separated skin.
采用了多种方法来破坏人皮肤的真皮-表皮连接（DEJ）。这些方法包括温热和热的磷酸盐缓冲盐水、胰蛋白酶、冷的1M盐、溴化钾以及用分散酶进行蛋白水解消化。在每种方法破坏DEJ后，测定基底膜成分（大疱性类天疱疮抗原、BM 600/奈辛、表皮整联配体蛋白、卡利宁、层粘连蛋白、IV型和VII型胶原蛋白（EBA抗原）以及连接蛋白）的定位和敏感性。我们发现，基底膜区蛋白BM 600/奈辛、表皮整联配体蛋白和卡利宁在通过冷盐、磷酸盐缓冲盐水和溴化钾在透明层处破坏的DEJ的真皮侧。用胰蛋白酶处理后未检测到BM 600/奈辛、表皮整联配体蛋白和卡利宁。相比之下，透明层中的另一种糖蛋白层粘连蛋白对所有这些操作均不敏感，但与破坏DEJ的皮肤的真皮侧共定位。我们还发现，将皮肤短暂暴露于56摄氏度来分离DEJ，为检测获得性大疱性表皮松解症（EBA）患者血清中的自身抗体提供了一种有用的底物。与盐分离的皮肤相比，热分离的皮肤可以在显著更短的时间内制备。
