Mechanisms of antibody-mediated lung Injury after blood transfusion

输血后抗体介导的肺损伤机制

• 批准号: 10318593
• 负责人: MARK ROBERTS LOONEY
• 金额: $ 57.73万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-01-01 至 2022-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10318593
• 关键词: Acute Lung Injury; Address; Affect; Affinity; Allogenic; Animals; Antibodies; Antibody Affinity; Antigen Targeting; Antigens; Area; Attention; Binding; Biochemical; Biochemistry; Biological Assay; Biology; Blood; Blood Platelets; Blood Transfusion; Blood donor; Cells; Cessation of life; Characteristics; Cloning; Complication; Consensus; Coupling; Defect; Development; Disease; Effectiveness; Effector Cell; Endothelium; Engineering; Evaluation; Event; Fc Receptor; Fresh Frozen Plasmas; Hemorrhage; Histocompatibility Antigens Class I; Hospitals; Human; IgG Receptors; IgG1; IgG4; Immune; Immune response; Immunoglobulin Class Switching; Immunoglobulin G; Immunologic Receptors; Incidence; Injury; Interleukin 2 Receptor Gamma; Knockout Mice; Knowledge; Lead; Leukocytes; Life; Ligation; Location; Lung; MHC Class I Genes; Malaria; Maps; Mediating; Mediator of activation protein; Medical; Modeling; Monoclonal Antibodies; Morbidity - disease rate; Mus; Pathogenesis; Pathogenicity; Pathology; Patients; Play; Population; Production; Property; Proteins; Recombinants; Role; Savings; Series; Severities; Site; Specificity; Surgical Blood Loss; System; Testing; Tissues; Transfusion; Traumatic injury; antibody engineering; base; blood product; cell type; cellular targeting; conditional knockout; cross reactivity; experience; experimental study; gene product; glycosylation; humanized mouse; immunoreaction; in vivo; lung injury; male; monocyte; mortality; mouse model; neutrophil; novel; prevent; receptor; receptor expression; selective expression; tool; transfusion related acute lung injury; translational approach

PROJECT SUMMARY/ABSTRACT Blood transfusions are life-saving for many patients with blood loss or blood production defects, but there are complications that limit their effectiveness and/or lead to medical sequelae. One such sequela is the development of acute lung injury, or transfusion-related acute lung injury (TRALI). TRALI has long been the number one cause of transfusion-related death and a major contributor to in-hospital morbidity. The pathogenesis of TRALI is incompletely understood, but there is consensus that TRALI is a classic example of antibody-mediated disease with a prominent role of HLA/MHC antibodies present in the donor blood product inciting lung injury in susceptible recipients. Indeed, we have developed a mouse model of TRALI based on transfusion of MHC Class I antibody into mice expressing the cognate MHCI antigen. In this application, we will use this model as a platform to investigate three specific aims. In Aim 1, we will determine the critical site of cognate antigen expression required to initiate TRALI by testing mice that are engineered to be deficient or sufficient in antigen expression in specific cellular locations. Novel mouse tools will be used to selectively delete Class I expression (B2mfl/fl) or selectively express allo-Class I protein (Con-Kd) on the endothelium or on specific immune cells. Using these complimentary mouse tools, we will definitively address the critical site of antigen expression required to initiate TRALI. In Aim 2, we will turn our attention to the biochemical characteristics of antibodies associated with the induction of TRALI. A single mouse monoclonal antibody against Class I (34-1- 2S) has been described to produce TRALI, whereas many other antibodies against the same antigen do not cause TRALI; it is unclear why this antibody is unique in its pathogenicity. We will test the hypotheses that antibody affinity, IgG subtype, glycosylation, and cross-reactivity each influence the induction of TRALI. These studies will utilize cutting-edge antibody engineering and binding and affinity assays to determine the properties of antibodies that impart pathogenicity in TRALI. In Aim 3, we will focus on the downstream events resulting from antibody engagement and recognition by immune cells. FcγRs are immune receptors that are the proximal trigger of antibody-mediated immune responses. Using a new mouse model that allows for the conditional deletion of activating FcγRs (Con-Fcγ-KO), we will determine the critical cells responsible for antibody recognition in TRALI. We will further determine if an inhibitory FcγR (FcγRII) influences TRALI severity. Finally, we will humanize the 34-1-2S antibody and infuse it into mice engineered to express human FcγRs in the absence of murine FcγRs. 34-1-2S will be expressed as each of the different human IgG subtypes (IgG1-IgG4) testing the hypothesis that IgG subtype affects TRALI in humans. The results of these experiments will definitively map the cells, antibody determinants, and receptors that are critical to TRALI induction. This knowledge will aid in our understanding of this serious complication of transfusion therapy and inform translational strategies to prevent or treat TRALI.

项目摘要/摘要 对于许多患有失血或血液产生缺陷的患者，输血是挽救生命的，但是有 限制其有效性和/或导致医学后遗症的并发症。这样的续集是 急性肺损伤或输血相关的急性肺损伤（TRALI）的发展。 Trali长期以来一直是 与输血相关死亡的第一原因，是院内发病率的主要贡献者。这 Trali的发病机理尚不完全理解，但是有共识，Trali是一个经典的例子 抗体介导的疾病，具有HLA/MHC抗体的重要作用 煽动敏感接受者的肺损伤。确实，我们基于 将MHC I类抗体输注到表达同源MHCI抗原的小鼠中。在此应用程序中，我们将 使用此模型作为调查三个特定目标的平台。在AIM 1中，我们将确定 通过测试设计为缺乏或 在特定细胞位置的抗原表达足够。新颖的鼠标工具将用于选择性删除 I类表达（B2MFL/FL）或选择性地表达内皮上或特定的Allo-Class I蛋白（CON-KD） 免疫细胞。使用这些免费的鼠标工具，我们将确定地解决抗原的关键位点 启动trali所需的表达。在AIM 2中，我们将把注意力转向生化特征 与Trali诱导相关的抗体。一种针对I类的小鼠单克隆抗体（34-1-- 2s）被描述为产生trali，而许多其他针对同一抗原的抗体则不 导致Trali；目前尚不清楚为什么这种抗体在致病性上是独特的。我们将测试假设 抗体亲和力，IgG亚型，糖基化和交叉反应性每种影响Trali的诱导。这些 研究将利用尖端的抗体工程和结合和亲和力修饰来确定特性 在Trali中赋予致病性的抗体。在AIM 3中，我们将重点放在下游事件上 来自免疫细胞的抗体参与和识别。 FcγR是免疫受体，是近端 抗体介导的免疫响应的触发。使用允许有条件的新鼠标模型 删除激活FcγR（CON-FCγ-KO），我们将确定负责抗体识别的关键细胞 在特拉利。我们将进一步确定抑制性FcγR（FcγRII）是否影响Trali的严重程度。最后，我们会的 人性化34-1-2S抗体，并将其注入以表达人FcγR的小鼠中 鼠FcγR。 34-1-2S将表示为每种不同的人IgG亚型（IgG1-igG4）测试 IgG亚型影响人类Trali的假设。这些实验的结果将一定地绘制 细胞，抗体确定剂和接收器对Trali诱导至关重要。这些知识将有助于我们 理解这种严重的翻译疗法并告知翻译策略以防止这种严重的并发症 或治疗Trali。

项目成果

An update of the transfusion-related acute lung injury (TRALI) definition.

输血相关急性肺损伤 (TRALI) 定义的更新。

• DOI: 10.1016/j.tracli.2019.05.007
• 发表时间: 2019
• 期刊: Transfusion clinique et biologique : journal de la Societe francaise de transfusion sanguine
• 作者: Vlaar,AlexanderPJ;Toy,Pearl;Fung,Mark;Looney,MarkR;Juffermans,NicoleP;Bux,Juergen;Bolton-Maggs,P;Peters,AnnaL;Silliman,ChristopherC;Kor,DarylJ;Kleinman,Steve
• 通讯作者: Kleinman,Steve