Complement Induces Inflammasome Assembly in Human Endothelium: Mechanisms and Consequences for Graft Rejection

补体诱导人内皮细胞炎症小体组装：移植物排斥的机制和后果

基本信息

批准号：
9609855
负责人：
Catherine Bingchan Xie
金额：
$ 2.93万
依托单位：
YALE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2018
资助国家：
美国
起止时间：
2018-06-01 至 2022-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9609855
关键词：
Acute Alleles Allogenic Allografting Antibodies Arteries Binding Biological Blood Vessels CASP1 gene Cell Adhesion Molecules Chronic Clathrin Cleaved cell Complement Complement Activation Complement Membrane Attack Complex Complex Data Deposition Development Disease Endocytosis Endosomes Endothelial Cells Endothelium Event Failure Family member Forms Controls Foundations Gene Expression Graft Rejection Heart failure Human Immune Immunodeficient Mouse Immunologics Immunology In Vitro Incidence Infiltration Inflammasome Inflammatory Injury Interferon Type II Interleukin-1 Interleukin-1 Receptors Interleukin-1 beta Isoantibodies Kidney Lead Link Liver Lung Measures Mediating Messenger RNA Modeling Organ Transplantation Pathogenesis Pathway interactions Patient-Focused Outcomes Phenotype Phosphorylation Phosphotransferases Physicians Process Production Protein Biosynthesis Protein Kinase Risk Role Savings Scaffolding Protein Scientist Secondary to Signal Pathway Signal Transduction Solid T cell response T memory cell T-Cell Activation T-Cell Proliferation T-Lymphocyte Therapeutic Uses Training Transplantation Up-Regulation Vascular Diseases Work allograft rejection base career cell fixing chemokine clinically relevant cytokine deep sequencing humanized mouse immune function immunogenicity improved in vitro Model in vivo mouse model new therapeutic target novel prevent receptor binding recruit response transplantation medicine

项目摘要

Solid organ transplantation, the best available treatment for end-stage kidney, liver, lung and heart failure, may fail due to chronic immunological rejection, often taking the form of allograft vasculopathy. Allograft vasculopathy results from recruitment to and activation within the graft vessel wall of IFN-γ-producing graft- reactive host T cells by graft ECs. The development of donor specific antibodies (DSA) that recognize non-self alleles of MHC molecules expressed by graft endothelial cells (ECs) and that fix complement is a major risk for allograft vasculopathy. Complement enhances this process by activating immune functions of the ECs that mediate both recruitment and activation of alloreactive T cells. Binding of high titer panel reactive antibody (PRA) from allo-sensitized transplant candidates, used to model DSA, deposits complement membrane attack complex (MAC) on human ECs, both in culture or in vessel grafts in immunodeficient mouse hosts, and induces gene expression of adhesion molecules and chemokines in a manner dependent upon MAC internalization and non-canonical NF-κB signaling. The mechanism(s) by which MAC potentiates T cell activation, measured as effector memory T cell proliferation and cytokine production in vitro or augmented vasculopathic changes in vivo is unknown. My preliminary data show that MAC induces formation of an active inflammasome in ECs, a previously undescribed phenomenon. Activated caspase-1 in the inflammasome processes pro-IL-1β to active IL-1β and mediates its release. Blocking caspase-1 activity or inhibiting IL-1 signaling with IL-1 receptor antagonist (IL-1Ra) blocks both downstream inflammatory gene expression in ECs secondary to canonical NF-κB activation and also blocks the augmentation of allogeneic T cell response in vitro. I hypothesize that complement activation of the inflammasome in ECs intensifies the host T cell response to graft arterial ECs by increasing local production of IL-1, potentiating allograft vasculopathy. In Specific Aim 1, I will characterize the MAC-induced inflammasome and determine the mechanisms linking MAC to inflammasome assembly. I will examine if kinases of non-canonical NF-κB signaling are linked to inflammasome assembly or IL-1 mRNA and protein synthesis. In Specific Aim 2, I will investigate the biological consequences of alloantibody-induced inflammasome activation in ECs on the alloreactive T cell response that drives allograft vasculopathy. I will determine if this response is also MAC-dependent and if mature IL-1 released from MAC-induced inflammasomes acts on ECs, T cells or both. I will characterize IL-1 dependent changes of the clonal repertoire and subsets of activated alloreactive effector memory T cells using TCR deep sequencing and FACS-analytic phenotyping and the effect of IL-1Ra or caspase-1 inhibition on the alloreactive T cell response in vitro using cultured human ECs. I will use well-developed humanized mouse models of allograft vasculopathy to assess the role of the EC inflammasome in vivo. Successful completion of this project may reveal novel targets for therapies to improve patient outcomes in transplant medicine.

固体器官移植，终末期肾脏，肝脏，肺和心脏的最佳治疗方法失败，可能是由于慢性免疫原理排斥而失败，通常采取同种异体移植血管病的形式。血管病来自募集到产生IFN-γ的移植物壁内的激活和激活通过移植EC的反应性宿主T细胞。识别非自我的供体特异性抗体（DSA）的开发由移植物内皮细胞（EC）表达的MHC分子等位基因和固定完成是主要风险同种异体血管病。补体通过激活EC的免疫功能来增强此过程介导同种异体T细胞的募集和激活。高滴度面板反应性抗体的结合（PRA）来自用于模拟DSA的同种敏感的移植候选者，沉积物补充膜攻击在培养物或容器移植物中，在人类EC上的复合物（MAC），免疫缺陷小鼠宿主和以依赖于MAC的方式诱导粘合分子和趋化因子的基因表达内在化和非典型的NF-κB信号传导。 MAC增强T细胞的机制激活，以效应子记忆T细胞的增殖和细胞因子的体外或增强作用测量体内的血管性变化尚不清楚。我的初步数据表明，MAC诱导了活动的形成 EC中的炎症体，这是一种以前未描述的现象。炎性体中激活的caspase-1 处理主动IL-1β并介导其释放。阻止caspase-1活性或抑制IL-1 IL-1受体拮抗剂（IL-1RA）的信号传导阻断了EC中的两个下游炎症基因表达继发于规范的NF-κB激活，也阻止了同种异体T细胞反应的增强我假设EC中炎症体的完成激活增强了宿主T细胞反应通过增加IL-1的局部产生，引起同种异体移植血管病。在特定目标中 1，我将表征MAC引起的炎症体，并确定将MAC连接到的机制炎症组件。我将检查非传统NF-κB信号的激酶是否与炎性体组装或IL-1 mRNA和蛋白质合成。在特定目标2中，我将研究生物学同种异体诱导的EC中炎症体激活的后果对同种反应性T细胞反应的影响驱动同类血管病。我将确定此响应是否也依赖于MAC，并且是否成熟IL-1 从MAC诱导的炎症中释放的作用于EC，T细胞或两者兼而有之。我会描述IL-1依赖性克隆曲目的变化和激活的同种异体效应记忆T细胞的子集使用TCR深测序和FACS分析表型以及IL-1RA或caspase-1抑制对同种异体反应性的影响使用培养的人类EC在体外进行T细胞反应。我将使用发达的人性化鼠标模型同种异体移植血管病以评估体内EC炎性体的作用。成功完成该项目可能会揭示用于改善移植药物患者预后的疗法的新靶标。