ISOLATION OF PEMPHIGUS ANTIGEN CDNA

寻常型天疱疮是一种严重的水疱性疾病，其特征是 循环中的免疫球蛋白与一种未知的表皮细胞抗原结合并引起 表皮脱落。在此之前，这种疾病都是致命的。 发现大剂量皮质类固醇治疗将导致患者病情缓解 大多数病人。然而，类固醇治疗有许多众所周知的并发症， 而一种更具体的疗法将具有很大的价值。 天疱疮抗原是一种细胞表面糖蛋白，其表达形式为 早在胎龄9周的时候，在某些表皮上的表皮细胞上 肿瘤，如A431，以及人羊膜上的。的大小 天疱疮抗原已知为210kD，由表皮免疫沉淀而成。 天疱疮细胞提取免疫球蛋白，但条带减少至130kD和80kD 在SDS-聚丙烯酰胺凝胶电泳法上。规模化提纯工业废水 到目前为止，抗原还没有用传统的方法完成。 我们建议从人羊膜中筛选出人的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 探测器。我们希望借此鉴定能够表达的细菌克隆 体外培养的天疱疮抗原。对这些克隆的分析将使我们能够 推导全部或部分天疱疮抗原的氨基酸序列，以 分离和研究其基因(S)及其表达调控 这种表皮细胞表面的糖蛋白。

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.