Okada N, Kitano Y, Miyagawa S, Sakamoto K, Steinberg M L
J Invest Dermatol. 1986 Apr;86(4):399-401. doi: 10.1111/1523-1747.ep12285666.
The expression and properties of pemphigoid antigen of SV40-transformed human keratinocytes were studied. By indirect immunofluorescence, SV40-transformed keratinocytes in passage 80-85 expressed the pemphigoid antigen as coarsely granular perinuclear fluorescence. To characterize this antigen, NP40 extracts of cells labeled with [14C]amino acids were immunoprecipitated using sera of 8 patients: bullous pemphigoid (6 patients), chronic localized pemphigoid (1 patient), and drug-induced lichen planus pemphigoides (1 patient). These immunoprecipitates were subjected to sodium dodecyl sulfate polyacrylamide gel electrophoresis and then fluorographed. All 8 sera precipitated a protein of Mr 240K, while normal human sera did not precipitate this protein. These results indicate that SV40-transformed human keratinocytes synthesize pemphigoid antigen, and that autoantibodies in the sera of pemphigoid patients with different clinical features identify the same antigen of Mr 240K in these cells.
研究了SV40转化的人角质形成细胞类天疱疮抗原的表达及特性。通过间接免疫荧光法,第80 - 85代的SV40转化角质形成细胞将类天疱疮抗原表达为粗大颗粒状的核周荧光。为了鉴定这种抗原,使用8例患者的血清对用[14C]氨基酸标记的细胞的NP40提取物进行免疫沉淀:大疱性类天疱疮(6例)、慢性局限性类天疱疮(1例)和药物性扁平苔藓类天疱疮(1例)。将这些免疫沉淀物进行十二烷基硫酸钠聚丙烯酰胺凝胶电泳,然后进行荧光自显影。所有8份血清均沉淀出一条分子量为240K的蛋白质,而正常人血清未沉淀出该蛋白质。这些结果表明,SV40转化的人角质形成细胞合成类天疱疮抗原,并且具有不同临床特征的类天疱疮患者血清中的自身抗体识别这些细胞中相同的分子量为240K的抗原。