Cloning and characterization of human pemphigus autoantibodies

描述（由申请人提供）： 该提案描述了一个为期五年的跨学科的职业发展计划，为医生科学家在学术皮肤病学。拟议计划的长期目标是继续发展和扩大主要研究者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。该科学计划的中心是天疱疮的发病机制，天疱疮是一种潜在的致命性起泡疾病，其中抗桥粒芯蛋白（Dsg）3和1的自身抗体，桥粒粘附分子，导致角质形成细胞粘附的丧失。本研究计划拟克隆和表征人寻常型天疱疮（PV）单克隆抗体（mAb），以确定PV中的致病性抗体队列。具体而言，所提出的实验旨在确定来自不同PV患者的抗Dsg mAb是否表现出共享的抗体基因使用和/或独特型，并使用这些mAb试剂来确定PV mAb诱导的棘层松解的机制。主要研究者在生物医学研究方面有15年的经验，目前正在宾夕法尼亚大学参加皮肤病学住院医师和研究培训计划。John R.斯坦利，医学博士，皮肤科系主任，和唐L。西格尔博士，医学博士，病理学和实验室医学系副主任将担任主要研究者科学和临床发展的共同导师。Stanley博士在天疱疮患者的临床护理方面拥有超过30年的经验，已经鉴定和表征了多种自身免疫性起泡疾病中涉及的抗原，并培养了许多博士后研究员，其中许多人已成为国内和国际学术皮肤病学的领导者。Siegel博士是国际知名的噬菌体展示专家，并指导临床单采单位，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一个独特的研究和临床培训的结合，从两个既定的和高度重视的医生科学家。本提案中概述的研究代表了对人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般范式。具体到PV，该项目将确定不同的人mAb如何促进疾病的病理学，开发有价值的人mAb试剂用于天疱疮和桥粒细胞粘附的研究，并可能导致这种危及生命的疾病的新型靶向治疗。

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.