Identification of human natural IgM that mediate ischemia-reperfusion injury

介导缺血再灌注损伤的人类天然 IgM 的鉴定

• 批准号: 7666780
• 负责人: Eric Garland Sheu
• 金额: $ 5.17万
• 依托单位: BRIGHAM AND WOMEN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-06-01 至 2010-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7666780
• 关键词: Adoptive Transfer; Antibodies; Antibody Repertoire; Antibody Specificity; Autoantigens; Autoimmunity; Autologous; Binding; Burn injury; Cells; Cloning; Complement; Data; Disease; Experimental Models; Goals; Host Defense; Human; Human Cloning; Human Identifications; Hybridomas; Immunoglobulin Genes; Immunoglobulin M; Immunoglobulins; Inflammatory; Injury; Intestines; Ischemia; Mediating; Mus; Myocardial Infarction; Operative Surgical Procedures; Outcome; Pathogenesis; Pathologic; Peritoneal; Reperfusion Injury; Reperfusion Therapy; Reverse Transcriptase Polymerase Chain Reaction; Rodent Model; Role; SCID Mice; Screening procedure; Skeletal Muscle; Skin Transplantation; Skin graft; Stroke; Testing; Tissues; chimeric antibody; human tissue; improved; reconstitution; therapeutic target; tool

DESCRIPTION (provided by applicant): Ischemia-reperfusion (I/R) injury is characterized by autologous inflammatory attack that is mediated by natural IgM antibody and complement. A single IgM clone produced by a murine peritoneal B1 cell hybridoma triggers reperfusion injury of skeletal muscle, intestine, and scald burns in rodent models. This natural IgM antibody binds exposed self-antigens on ischemic tissue and activates complement, leading to tissue injury. An analogous pathologic mechanism of I/R injury in humans has been proposed but is unproven. Preliminary data from our group demonstrate that adoptive transfer of human IgM into RAG2 -/-mice reconstitutes reperfusion injury of murine and human tissue. The goal of this proposal is to identify human IgM involved in ischemia-reperfusion injury. We hypothesize that human peritoneal B1 cells harbor one or several antibody determinants that can bind self antigen and initiate the inflammatory cascade leading to I/R injury. We will test this hypothesis through the following specific aims: (1) cloning of immunoglobulin molecules expressed by human peritoneal B1 cells using single cell RT-PCR; (2) expression and screening of human natural IgM for reconstitution of reperfusion injury in antibody deficient RAG2 -/- mice and of burn injury in SCID mice bearing human skin grafts. Identification of a human natural antibody involved in ischemia-reperfusion injury will provide potential therapeutic targets for down regulating I/R injury and thus improving outcomes in stroke, heart attacks, and surgery. Characterization of the human natural antibody repertoire would also be an important tool for understanding B1 cells and their role in host defense and autoimmunity.

描述(申请人提供)：缺血-再灌注(I/R)损伤的特征是由天然的IgM抗体和补体介导的自体炎症发作。由小鼠腹膜B1细胞杂交瘤产生的单个IgM克隆在啮齿动物模型中触发骨骼肌、肠道和烫伤的再灌注损伤。这种天然的IgM抗体与缺血组织上暴露的自身抗原结合，激活补体，导致组织损伤。已经提出了一种类似的人类I/R损伤的病理机制，但尚未得到证实。本课题组的初步数据表明，过继将人IgM转移到RAG2-/-小鼠体内，可以重建小鼠和人组织的再灌注损伤。 本研究的目的是鉴定与缺血再灌注损伤相关的人IgM。我们假设人腹膜B1细胞含有一个或几个抗体决定簇，可以结合自身抗原，启动导致I/R损伤的炎症级联反应。我们将通过以下特定目标来验证这一假设：(1)使用单细胞RT-PCR克隆人腹膜B1细胞表达的免疫球蛋白分子；(2)表达和筛选人天然IgM，用于抗体缺陷RAG2-/-小鼠的再灌注损伤和携带人皮肤移植的SCID小鼠的烧伤损伤的重建。 鉴定一种与缺血-再灌注损伤有关的人类天然抗体将为下调I/R损伤提供潜在的治疗靶点，从而改善中风、心脏病发作和手术的预后。人类天然抗体库的特征也将是了解B1细胞及其在宿主防御和自身免疫中的作用的重要工具。