PATHOGENESIS OF EXPERIMENTAL GLOMERULONEPHRITIS

本研究的目的是分析抗体的作用机制， 诱导的肾小球肾炎（GN）的大鼠时，靶抗原 决定簇是一个单一的明确定义的结构，通过使用单克隆 （杂交瘤）抗体作为特异性探针。 我们将生产老鼠 针对大鼠肾小球基底膜（GBM）蛋白的杂交瘤， 培养的肾小球细胞，并初步确定的性质， 得到的单克隆抗体（免疫球蛋白的类别和亚类， 固定补体的能力和良好的特异性）。 不同单克隆 将抗体制剂被动给予大鼠， 肾小球功能和形态异常。 致肾炎和 将比较非致肾炎抗体的量， 与肾脏的结合、周转率、结合定位和 目的是为了确定异源的性质， 这些抗体对于被动GN的诱导至关重要。 目标 致肾炎和非致肾炎抗体的抗原将 从溶解的GBM制备物中纯化，使用抗体作为 特异性免疫吸附剂。 这些抗原将被表征，并且其 通过抗体结合确定在肾脏中的定位。 最后， 将尝试通过以下方法诱导主动的自身免疫形式的GN 用纯化的自体抗原免疫动物， 这种自身免疫性GN的发病机制将是 通过体外试验和过继转移实验分析。

The goal of this research project is to analyze the mechanisms of antibody induced glomerulonephritis (GN) in rats when the target antigenic determinant is a single well-defined structure, by using monoclonal (hybridoma) antibodies as specific probes. We will produce mouse hybridomas against rat glomerular basement membrane (GBM) proteins and cultured glomerular cells, and initially define the properties of the resultant monoclonal antibodies (class and subclass of immunoglobulin, ability to fix complement and fine specificity). Different monoclonal antibody preparations will be passively administered to rats to produce functional and morphologic glomerular abnormalities. Nephritogenic and non-nephritogenic antibodies will be compared with respect to quantity of binding to kidneys, turnover rates, localization of binding and ability to fix complement with a view to defining the properties of heterologous antibodies that are critical for the induction of passive GN. The target antigens for nephritogenic and non-nephritogenic antibodies will be purified from solubilized GBM preparations, using the antibodies as specific immunoabsorbents. These antigens will be characterized, and their localization in the kidneys determined by antibody binding. Finally, attempts will be made to induce an active, auto-immune form of GN by immunizing animals with purified autologous antigens, and the immune mechanisms involved in the pathogenesis of this anto-immune GN will be analyzed by in vitro tests and adoptive transfer experiments.