PATHOGENESIS OF MEMBRANOUS GLOMERULONEPHROPATHY

描述（改编自研究者摘要）：研究者 长期目标是了解发病机制 膜性肾小球肾病（MGN）。 他们将与公认的 MGN、大鼠活动性Heymann肾炎（HN）实验模型， 将他们的研究集中在假定的自身抗原gp 330上， 参与发病机制。 其具体目标是：（A） 确定B细胞的位置和结构（氨基酸序列） 参与疾病的gp 330表位。 为了实现这一目标，他们将 分离与自身抗体反应gp 330蛋白水解片段， 对片段进行微测序，将其定位在gp 330序列中，克隆到 通过PCR扩增制备相应cDNA的表达载体， 检测表达的融合蛋白的自身抗体反应性， 免疫印迹和ELISA，并通过免疫大鼠检测免疫原性。 他们 将缩小连续和不连续表位的位置， 通过分析PCR产生的较小克隆， 跨越表位区域，并鉴定 自身抗体反应性线性表位和可能的序列 参与不连续表位，并测试其致病潜力 所有已识别的表位。 B）测试鉴定的序列， 在活性HN中的致耐受性和治疗潜力， 较大量的推定肽或蛋白质片段， 将这些肽/蛋白质片段植入动物体内， 参数 这些研究的数据将提供重要的新信息 参与活动性HN的gp 330的B细胞表位的结构。 这些知识将导致1）更好地了解发病机制 这种疾病在分子水平上，2）可能是它的治疗， 特异性免疫调节。

DESCRIPTION (Adapted from Investigator's Abstract): The investigators' long-term goal is to understand the mechanisms involved in the pathogenesis of membranous glomerulonephropathy (MGN). They will work with the accepted experimental model of MGN, active Heymann Nephritis (HN) of rat, and concentrate their studies on the putative autoantigen, gp330, a key participant in the mechanisms of pathogenesis. Their specific aims are: A) Identify the location and structure (amino acid sequence) of the B-cell epitopes of gp330 involved in the disease. To achieve this they will isolate gp330 proteolytic fragments reactive with autoantibodies, microsequence the fragments, localize them in gp330 sequence, clone into expression vectors the corresponding cDNA's prepared by PCR amplification, test expressed fusion proteins for autoantibodies-reactivity by immunoblotting and ELISA, and for immunogenicity by immunizing rats. They will narrow down the location of the continuous and discontinuous epitope(s) by analyzing smaller clones produced by PCR, synthesize overlapping peptides spanning the epitope regions and identify the precise amino acid sequence of autoantibodies-reactive linear epitope(s) and the sequences possibly involved in the discontinuous epitopes, and test the pathogenic potential of all of the identified epitopes. B) Test the identified sequences for tolerogenic and therapeutic potential in active HN by synthesizing the putative peptide(s) or protein fragments in larger quantities, administering these peptide/protein fragments into animals, and then assessing the disease parameters. Data from these studies will provide important new information on the structure of the B cell epitopes of gp330 involved in active HN. This knowledge will lead to 1) a greater understanding of the pathogenesis of this disease at the molecular level and, 2) possibly its treatment with specific immunomodulation.