Ebihara T, Hashimoto T, Kudoh J, Gamou S, Shimizu N, Nishikawa T
Department of Dermatology, Keio University School of Medicine, Tokyo, Japan.
J Invest Dermatol. 1993 Feb;100(2):176-9. doi: 10.1111/1523-1747.ep12462799.
There has been controversy concerning the nature of the bullous pemphigoid (BP) antigen: immunoprecipitation identified BP antigen as a single, unique 230-kDa protein, whereas immunoblot analysis showed multiple antigen molecules, mainly 230- and 170-kDa proteins. In this study, to further characterize the 170-kDa protein, we have examined whether the 170-kDa protein is detected by immunoprecipitation. Extracts of human squamous cell carcinoma cells revealed the 170-kDa protein with immunoblot analysis. Although the conventional immunoprecipitation detected only the 230-kDa protein, some BP sera that detected the 170-kDa protein with immunoblotting also precipitated the 170-kDa protein with our modified immunoprecipitation, in which the cells were extracted with 1% sodium dodecylsulfate (SDS) buffer and reacted with the sera under reduced SDS concentration. The 170-kDa protein-specific BP sera clearly showed hemidesmosomal plaque staining with immunofluorescence of cultured cells. These results indicate that the 170-kDa protein is indeed one of the BP antigens and that the 230- and 170-kDa BP antigens are integrated in different ways in hemidesmosomes.
关于大疱性类天疱疮（BP）抗原的性质一直存在争议：免疫沉淀将BP抗原鉴定为一种单一、独特的230 kDa蛋白质，而免疫印迹分析显示有多种抗原分子，主要是230 kDa和170 kDa的蛋白质。在本研究中，为了进一步表征170 kDa蛋白质，我们检测了免疫沉淀是否能检测到170 kDa蛋白质。人鳞状细胞癌细胞提取物经免疫印迹分析显示出170 kDa蛋白质。尽管传统免疫沉淀仅检测到230 kDa蛋白质，但一些能通过免疫印迹检测到170 kDa蛋白质的BP血清，在我们改良的免疫沉淀中也沉淀出了170 kDa蛋白质，改良方法是用1%十二烷基硫酸钠（SDS）缓冲液提取细胞，并在降低的SDS浓度下与血清反应。170 kDa蛋白质特异性BP血清在培养细胞的免疫荧光检测中明显显示半桥粒斑块染色。这些结果表明，170 kDa蛋白质确实是BP抗原之一，并且230 kDa和170 kDa的BP抗原以不同方式整合在半桥粒中。
