Human Autoantibodies in Anti-GBM Disease

抗 GBM 疾病中的人类自身抗体

• 批准号: 6743164
• 负责人: ANNE M CHRISTENSEN
• 金额: $ 13.07万
• 依托单位: CHILDREN'S HOSP OF PHILADELPHIA
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-05-01 至 2008-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6743164
• 关键词: antigen antibody reaction; autoantibody; autoimmune disorder; basement membrane; conformation; disease /disorder model; enzyme linked immunosorbent assay; epitope mapping; glomerulonephritis; human genetic material tag; immunologic substance development /preparation; immunopathology; laboratory mouse; monoclonal antibody; protein sequence; protein structure function; renal glomerulus; western blottings

DESCRIPTION (provided by applicant):Anti glomerular basement membrane (GBM) disease is an autoimmune disease mediated by antibodies against the GBM. The usual clinical picture is rapidly progressive glomerulonephritis (RPGN) resulting in end stage renal disease. These patients consequently suffer a significant reduction in quality of life, due to life-long need for renal replacement therapy. While significant knowledge currently exist about the structural basis of the antigen (Ag) of human anti-GBM disease, alpha3(IV) NCI collagen (alpha3(IV)); the pathogenesis of this autoimmune disease has yet to be fully elucidated. Although putative antigenic epitopes have been identified, little is known about the origin, structure and conformation of pathogenic human anti-GBM antibodies (Ab) that recognize these epitopes.The studies proposed here seek to identify how autoantibodies against alpha3(IV) initiate nephritis through binding to the GBM. Using the XenoMouseII (XMII) model of human anti-GBM disease that has been developed in our laboratory, I propose to generate human monoclonal antibodies (mAb), directed towards 2 putatively pathogenically relevant epitopes on Antibodies against the GBM. The usual clinical picture is rapidly progressive glomerulonephritis (RPGN) resulting in end stage renal disease. These patients consequently suffer a significant reduction in quality of life, due to life-long need for renal replacement therapy. While significant knowledge currently exist about the structural basis of the antigen (Ag) of human anti-GBM disease, alpha3(IV) NCI collagen (alpha3(IV)); the pathogenesis of this autoimmune disease has yet to be fully elucidated. Although putative antigenic epitopes have been identified, little is known about the origin, structure and conformation of pathogenic human anti-GBM antibodies (Ab) that recognize these epitopes. The studies proposed here seek to identify how autoantibodies against alpha3(IV) initiate nephritis through binding to the GBM. Using the XenoMouseII (XMII) model of human anti-GBM disease that has been developed in our laboratory, I propose to generate human monoclonal antibodies (mAb), directed towards 2 putatively pathogenically relevant epitopes on alpha3(IV) (Aim 1). The specificity and affinity with which these mAb bind alpha3(IV) in vitro will be determined (Aim 2), and correlated to their pathogenicity (Aim 3). Sequencing of the hypervariable region of the pathogenic mAb will allow prediction of the 3-dimensional structure of the Ag-binding region. Subsequently, this information should enable me to determine which amino acid residues in the hypervariable region are essential for the interaction with alpha3(IV) (Aim 2) and the disease producing autoAb (Aim 3). These studies should help provide insights into the basic pathophysiology of anti-GBM disease. Additionally, these results may aide in the design and evaluation of effective therapies (e.g., small molecules) for treatment of this devastating disease, in the future.The K08 mechanism will enable me to undertake mentored research training in basic immunology and the pathogenesis of autoimmune glomerulonephritis. These studies will be conducted in Dr. Michael P. Madaio's laboratory at the University of Pennsylvania. The training and studies will greatly facilitate my career goals of working as a physician/scientist in an academic setting.

描述（申请人提供）： 抗肾小球基底膜（GBM）疾病是一种由抗GBM抗体介导的自身免疫性疾病。通常的临床表现是快速进展性肾小球肾炎（RPGN），导致终末期肾病。由于终生需要肾脏替代治疗，这些患者的生活质量显着下降。虽然目前对人类抗 GBM 疾病的抗原 (Ag) α3(IV) NCI 胶原蛋白 (α3(IV)) 的结构基础有了重要的了解；这种自身免疫性疾病的发病机制尚未完全阐明。虽然推定的抗原表位已被识别，但人们对识别这些表位的致病性人类抗 GBM 抗体 (Ab) 的起源、结构和构象知之甚少。本文提出的研究旨在确定抗 α3(IV) 的自身抗体如何通过以下方式引发肾炎：与 GBM 结合。使用我们实验室开发的人类抗 GBM 疾病的 XenoMouseII (XMII) 模型，我建议生成人类单克隆抗体 (mAb)，针对 GBM 抗体上的 2 个假定的致病相关表位。通常的临床表现是快速进展性肾小球肾炎（RPGN），导致终末期肾病。由于终生需要肾脏替代治疗，这些患者的生活质量显着下降。虽然目前对人类抗 GBM 疾病的抗原 (Ag) α3(IV) NCI 胶原蛋白 (α3(IV)) 的结构基础有了重要的了解；这种自身免疫性疾病的发病机制尚未完全阐明。尽管已经鉴定出假定的抗原表位，但对于识别这些表位的致病性人类抗 GBM 抗体 (Ab) 的起源、结构和构象知之甚少。本文提出的研究旨在确定抗 alpha3(IV) 自身抗体如何通过与 GBM 结合引发肾炎。使用我们实验室开发的人类抗 GBM 疾病的 XenoMouseII (XMII) 模型，我建议生成人类单克隆抗体 (mAb)，针对 alpha3(IV) 上 2 个假定的致病相关表位（目标 1）。将确定这些 mAb 在体外结合 alpha3(IV) 的特异性和亲和力（目标 2），并与其致病性相关（目标 3）。对致病性 mAb 的高变区进行测序将能够预测 Ag 结合区的 3 维结构。随后，这些信息应该使我能够确定高变区中的哪些氨基酸残基对于与 alpha3(IV)（目标 2）和产生自身抗体的疾病（目标 3）的相互作用至关重要。这些研究应该有助于深入了解抗 GBM 疾病的基本病理生理学。此外，这些结果可能有助于将来设计和评估治疗这种破坏性疾病的有效疗法（例如小分子）。K08 机制将使我能够接受基础免疫学和自身免疫发病机制方面的指导研究培训肾小球肾炎。这些研究将在宾夕法尼亚大学 Michael P. Madaio 博士的实验室进行。培训和学习将极大地促进我在学术环境中担任医生/科学家的职业目标。