Cloning and characterization of human pemphigus autoantibodies

描述(由申请人提供)： 这项建议描述了一项为期五年的跨学科职业发展计划，该计划面向皮肤科学术领域的内科科学家。拟议计划的长期目标是继续发展和扩大首席研究员的研究和临床专业知识，特别关注皮肤自身免疫性起泡疾病的原因和治疗。科学计划集中在天疱疮的发病机制上，天疱疮是一种潜在的致命水泡性疾病，在这种疾病中，抗桥粒黏附分子桥粒蛋白3和1的自身抗体会导致角质形成细胞黏附丧失。该研究计划建议克隆和鉴定人类寻常型天疱疮(PV)的单抗，以确定PV中的致病抗体队列。具体地说，本实验旨在确定来自不同PV患者的抗DSG单抗是否表现出共同的抗体基因使用和/或独特型，并使用这些单抗试剂来确定PV单抗诱导棘层松解的机制。这位首席研究员在生物医学研究方面有15年的经验，目前在宾夕法尼亚大学参加皮肤病住院医师和研究培训计划。皮肤科主任约翰·R·斯坦利医学博士和病理学和实验室医学系副主任唐·L·西格尔博士将共同指导首席研究员的科学和临床发展。斯坦利博士在天疱疮患者的临床护理方面拥有30多年的经验，识别和表征了与多种自身免疫性起泡疾病有关的抗原，并培训了许多博士后研究员，其中许多人已成为国内和国际皮肤病学学术领域的领军人物。西格尔博士是国际知名的噬菌体展示专家，并领导临床分离单位，在那里他监督许多严重或难治性PV患者的治疗。拟议的跨学科计划提供了两位久负盛名的内科科学家的研究和临床培训的独特组合。这项建议中概述的研究代表了对人类PV单克隆自身抗体的第一次详细的遗传和功能分析，并可能为体液介导的自身免疫性疾病的研究建立一个通用的范例。针对PV，该项目将定义不同的人类mAb如何在疾病的病理中做出贡献，开发有价值的人类mAb试剂来研究天疱疮以及桥粒细胞黏附，并可能导致这种威胁生命的疾病的新的靶向治疗。

DESCRIPTION (provided by applicant): This proposal describes a five year interdisciplinary career development program for a physician scientist in academic dermatology. The long-term objectives of the proposed program are to continue to develop and expand the principal investigator's research and clinical expertise, with special focus on the causes and treatment of autoimmune blistering diseases of the skin. The scientific program centers on mechanisms of pathogenesis in pemphigus, a potentially fatal blistering disorder in which autoantibodies against desmoglein (Dsg) 3 and 1, desmosomal adhesion molecules, cause loss of keratinocyte cell adhesion. The research plan proposes to clone and characterize human pemphigus vulgaris (PV) monoclonal antibodies (mAbs) in order to define the pathogenic antibody cohort in PV. Specifically, the proposed experiments aim to determine whether anti-Dsg mAbs from different PV patients demonstrate shared antibody gene usage and/or idiotypes, and to use these mAb reagents to determine mechanisms of acantholysis induced by PV mAbs. The principal investigator has 15 years experience in biomedical research and is currently enrolled in a dermatology residency and research training program at the University of Pennsylvania. John R. Stanley, M.D., chairman of the Department of Dermatology, and Don L. Siegel, Ph.D., M.D., vice-chairman of the Department of Pathology and Laboratory Medicine, will serve as co-mentors for the principal investigator's scientific and clinical development. Dr. Stanley has over 30 years experience in the clinical care of pemphigus patients, has identified and characterized antigens involved in multiple autoimmune blistering disorders, and has trained numerous postdoctoral fellows, many of whom have become leaders in academic dermatology both nationally and internationally. Dr. Siegel is an internationally renowned expert in phage display and directs the clinical apheresis unit, where he supervises the treatment of many patients with severe or refractory PV. The proposed interdisciplinary program offers a unique combination of research and clinical training from two established and highly regarded physician scientists. The studies outlined within this proposal represent the first detailed genetic and functional analysis of human PV monoclonal autoantibodies and may establish a general paradigm for the study of humorally mediated autoimmune disease. Specific to PV, the project will define how different human mAbs contribute to the pathology of disease, develop valuable human mAb reagents for the study of pemphigus as well as desmosomal cell adhesion, and potentially lead to novel targeted therapies for this life-threatening disorder.