MONOCLONAL ANTIBODIES TO BULLOUS PEMPHIGOID ANTIGENS

大疱性类天疱疮是一种自身免疫性疾病，其中自身抗体 它们本身也与致病有关。我们的目标是 研制抗人免疫球蛋白相关抗原的单抗 大疱性类天疱疮。长期目标是制定一项计划，在该计划中 单抗可以用来阐明这种疾病和 通过抑制抗独特型进行免疫调节。为了实现我们的目标 主要目标是，杂交瘤融合将与来自 用类天疱疮抗原免疫的大鼠和小鼠，或用来自 精选类天疱疮患者。Fusion合作伙伴将由现有的 鼠源性或人源性的恶性细胞系。由此产生的 分泌表皮抗原抗体的杂交瘤将被鉴定 通过间接免疫荧光法。这些细胞系将被克隆以生产 单克隆抗体。这些单抗将被鉴定为 通过间接免疫荧光检测其组织特异性，并对其 免疫印迹技术对表皮表位的反应性。这个 单抗的免疫化学性质也将 下定决心。这些属性将包括等电点和表观 分子量(通过双向聚丙烯酰胺凝胶电泳法)， 类和子类。这些单抗也将用于 抗原的提纯及其对生物活性的探索 发病机制。后者将涉及一种体外试验 表皮细胞附着在固体底物上。单克隆抗体, 单独或组合，将与表皮基底细胞孵育 停职。抑制细胞与底物的附着 与抗体孵育，可提示和累及大疱病 类天疱疮抗原(S)与表皮基底细胞-底物相互作用。

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.