ISOLATION OF PEMPHIGUS ANTIGEN CDNA

寻常天疱疮是一种严重的起泡性疾病， 与未知表皮细胞抗原结合并导致 表皮脱落 这种疾病在1990年以前是致命的。 发现高剂量皮质类固醇治疗会导致 大多数病人 然而，类固醇治疗有许多众所周知的并发症， 更具体的治疗方法会很有价值 天疱疮抗原是细胞表面糖蛋白，其表达为 早在9周胎龄的表皮细胞上，在一些表皮细胞上， 癌如A431，和在人结肠癌细胞膜上。 的大小 天疱疮抗原已知为210 Kd，通过免疫沉淀表皮 细胞提取物与天疱疮IgG，但减少到130 Kd和80 Kd带 SDS-聚丙烯酰胺凝胶电泳 大规模纯化 迄今为止还没有使用常规方法完成抗原的制备。 我们拟从人羊膜中筛选人cDNA表达文库 和胎盘信使RNA，使用纯化的标记天疱疮抗体作为 探针 我们希望借此鉴定出表达 天疱疮抗原体外研究。 对这些克隆体的分析将使我们能够 推导天疱疮抗原的全部或部分的氨基酸序列， 分离和研究其基因，并研究其表达和调控， 这种表皮细胞表面糖蛋白。

Pemphigus vulgaris is a severe blistering disease characterized by a circulating IgG which binds to an unknown epidermal cell antigen and causes epidermal disadherance. The disease was uniformly fatal prior to the discovery that high dose corticosteroid therapy would induce a remission in most patients. However, steroid therapy has many well known complications, and a more specific therapy would be of great value. Pemphigus antigen is a cell surface glycoprotein which is expressed as early as 9 weeks gestational age on epidermal cells, on some epidermal carcinoma such as A431, and on human amnionic membranes. The size of pemphigus antigen is known to be 210 Kd by immunoprecipitation of epidermal cell extracts with pemphigus IgG but is reduced to 130 Kd and 80 Kd bands on SDS-polyacrylamide gel electrophoresis. Large scale purification of the antigen has not been accomplished to date using conventional methods. We propose to screen a human cDNA expression library made from human amnion and placenta messenger RNA using purified labeled pemphigus antibody as a probe. We hope thereby to identify bacterial clones which express pemphigus antigens in vitro. Analysis of such clones would enable us to derive the amino acid sequence for all or part of the pemphigus antigen, to isolate and study its gene(s) and to study the expression and regulation of this epidermal cell surface glycoprotein.