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.

项目总结/摘要 对于许多失血或血液生成缺陷的患者来说，输血是挽救生命的方法， 限制其有效性和/或导致医学后遗症的并发症。一个这样的后遗症是 急性肺损伤或输血相关急性肺损伤（TRALI）的发展。TRALI一直是 输血相关死亡的头号原因和院内发病率的主要贡献者。的 TRALI的发病机制尚不完全清楚，但有一个共识，TRALI是一个经典的例子， 抗体介导的疾病，供体血液制品中存在的HLA/MHC抗体发挥着重要作用 在易感受者中引发肺损伤。事实上，我们已经开发了TRALI的小鼠模型， 将MHC I类抗体输注到表达同源MHC I抗原的小鼠中。在这个应用程序中，我们将 以这个模型为平台，研究三个具体目标。在目标1中，我们将确定 通过测试经工程改造为缺陷或缺失的小鼠来启动TRALI所需的同源抗原表达， 在特定的细胞位置足够的抗原表达。新颖的鼠标工具将用于选择性删除 I类表达（B2 mfl/fl）或选择性表达内皮上或特异性表达I类蛋白（Con-Kd）。 免疫细胞使用这些互补的小鼠工具，我们将明确地解决抗原的关键位点， 启动TRALI所需的表达。在目标2中，我们将把注意力转向以下生物化学特征： 与TRALI诱导相关的抗体。一种抗I类（34-1- 34）的单鼠单克隆抗体， 2S）被描述为产生TRALI，而针对相同抗原的许多其他抗体不产生TRALI 导致TRALI;目前还不清楚为什么这种抗体在其致病性方面是独特的。我们将测试假设， 抗体亲和力、IgG亚型、糖基化和交叉反应性各自影响TRALI的诱导。这些 研究将利用尖端的抗体工程和结合及亲和力测定来确定这些特性， 在TRALI中赋予致病性的抗体。在目标3中，我们将重点关注下游事件， 免疫细胞的抗体结合和识别。Fcγ R是免疫受体， 引发抗体介导的免疫反应。使用一种新的小鼠模型， 删除活化Fcγ R（Con-Fcγ-KO），我们将确定负责抗体识别的关键细胞 在TRALI。我们将进一步确定抑制性FcγR（FcγRII）是否影响TRALI严重程度。最后我们将 人源化34-1-2S抗体并将其输注到经工程改造以在不存在人Fcγ R的情况下表达人Fcγ R的小鼠中。 鼠FcγRs。34-1-2S将表示为检测抗体的每种不同人IgG亚型（IgG 1-IgG 4）。 假设IgG亚型影响人类TRALI。这些实验的结果将明确地描绘出 细胞、抗体决定簇和受体对TRALI诱导至关重要。这些知识将有助于我们 了解这种严重的输血治疗并发症，并告知翻译策略，以防止 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