MONOCLONAL ANTIBODIES TO BULLOUS PEMPHIGOID ANTIGENS

大疱性类天疱疮是一种自身免疫性疾病， 它们本身也与病原体有关。 目标是 开发针对与以下相关的假定抗原的单克隆抗体： 大疱性类天疱疮 长期目标是制定一项计划， 单克隆抗体可用于阐明疾病 通过抑制性抗独特型进行免疫调节。 实现我们 主要目标是，将用来自以下的脾细胞进行杂交瘤融合： 用类天疱疮抗原免疫的大鼠和小鼠，或用来自 选择类天疱疮患者。 融合合作伙伴将包括已建立的 鼠或人来源的恶性细胞系。 所得 将鉴定分泌表皮抗原抗体杂交瘤 间接免疫荧光法。 这些细胞系将被克隆以产生 克隆抗体 将对单克隆抗体进行以下表征： 通过间接免疫荧光检测其组织特异性， 通过免疫印迹技术检测对表皮表位的反应性。 的 单克隆抗体的免疫化学性质也将被 测定 这些性质将包括等电点和表观 分子量（通过二维聚丙烯酰胺凝胶电泳）， class和subclass。 单克隆抗体也将用于 抗原的纯化和生物活性的探索 发病机制。 后者将涉及体外测定， 表皮细胞附着于固体基质。 单克隆抗体， 将单独或组合地与表皮基底细胞一起孵育 停职 抑制细胞附着到基底上， 与抗体孵育，可能表明和参与大疱性 类天疱疮抗原与表皮基底细胞-基质相互作用。

Bullous pemphigoid is an autoimmune disease in which the autoantibodies themselves have been implicated in pathogenensis. The objective is to develop monoclonal antibodies to the putative antigens concerned with bullous pemphigoid. Long range goals are to develop a program in which the monoclonal antibodies can be utilized for elucidation of the disease and for its immunoregulation by suppressive anti-idiotypes. To achieve our primary goal, hybridoma fusions will be performed with spleen cells from rats and mice immunized with pemphigoid antigens, or with lymphocytes from selected pemphigoid patients. Fusion partners will consist of established malignant cell lines of either murine or human origin. The resulting hybridomas that secrete antibody to epidermal antigens will be identified by indirect immunofluorescence. These cell lines will be cloned to produce monoclonal antibodies. The monoclonal antibodies will be characterized for their tissue specificity by indirect immunofluorescence and for their reactivity to epidermal epitopes by immunoblotting techniques. The immunochemical properties of the monoclonal antibodies will also be determined. These properties will include isoelectric point and apparent molecular weight (by 2-dimensional polyacrylamide gel electrophoresis), class, and subclass. The monoclonal antibodies will also be useful for purification of antigen and for the exploration of the biological mechanisms of pathogenesis. The latter will involve an in vitro assay for epidermal cell attachment to a solid substrate. Monoclonal antibodies, individually or in combination, will be incubated with epidermal basal cell suspensions. Inhibition of cell attachment to the substrate following incubation with antibody, may indicate and involvement of the bullous pemphigoid antigen(s) with the epidermal basal cell-substrate interaction.