Immunosurveillance and Immunopathology Core

项目摘要 免疫监测和免疫病理核心(IIC)通过对来自中国的样本进行机制研究 这项申请的3个项目将发挥协同科学作用，重点是移植。 宽容，这是本计划的中心焦点。IIC将进行分析免疫变化的化验 方案1、方案2和方案3中提出的耐受方案引起的外周循环、移植物和脾。 移植耐受实验室(TTL)制定了一系列标准操作程序(SOP) 为了分析NHP移植受者的供者特异性免疫反应，这些免疫反应将在所有 协调U19项目中所有项目的分析建议。核心已经优化了， 验证并采用了几种尖端技术和管道进行生物信息数据分析 跨多个组织隔间的复杂数据集。三个国家的样本的综合免疫分析 IIC的项目将有助于避免化验和分析的差异，从而统一结果， 将有助于在一个模型中获得洞察力，以便在其他项目中协同部署。IIC将执行 以下列出的四个具体目标中概述的分析，并进行生物信息学和数据分析管道 评估项目1、2和3中采用的不同耐受方案所引起的免疫变化。 目的1：用高维细胞免疫荧光技术研究肾移植受者免疫功能的变化 诱导和维持耐受性过程中循环免疫细胞的特征。目标2：生成和验证 使用单细胞TCRseq跟踪供者抗原特异性的CD4+和CD8+T细胞的TCR特征。目标3：定义 MHC-II类四聚体和单细胞ATAC诱导的供者抗原特异性CD4+T细胞的去向和功能 移植肾耐受和排斥的系列外周血单个核细胞的RNA测序 收件人。目的4：分析移植肾、脾和淋巴中免疫细胞的时空组织 通过数字空间轮廓在转录水平上的节点和通过MIBI-TOF在蛋白质水平上。分析 多个纵向组织隔间，以生成一个大的、但高度受控的数据集。我们是故意的 将这种多组学方法应用于系统级别的优势并行识别，而不是顺序识别 移植耐受的免疫机制分析。三个项目的样本分析 利用不同的耐受方案将提供新的免疫学见解，以诱导和维持 宽容。此外，跨项目的分析性能支持横向知识转移，以确定 共同性，独立于策略，揭示免疫耐受背后的主要特征，以推进 迈向成功的临床应用。IIC将定期与3个项目的绩效指标和 管理核心，以设计、执行、分析和解决本申请中提出的研究。

Project Summary The Immunosurveillance and Immunopathology Core (IIC) by performing mechanistic studies on samples from the 3 projects of this application will serve a synergistic scientific role with strong emphasis on transplant tolerance, the central focus of this Program. The IIC will perform assays to analyze the immune alterations in peripheral circulation, graft and spleen induced by the tolerance regimens proposed in Projects 1, 2 and 3. The Transplantation Tolerance Laboratory (TTL) has developed a series of Standard Operating Procedures (SOPs) to analyze the donor-specific immune responses in NHP transplant recipients that will be shared across all the projects to harmonize the analyses proposed across all the projects in the U19 program. The Core has optimized, validated, and adopted several cutting-edge technologies and pipelines for bioinformatic data analysis of this complex dataset across multiple tissue compartments. Comprehensive immune analyses of samples from the 3 projects by the IIC will aid in avoidance of assay and analysis variations resulting in harmonization of results that will aid in insights obtained in one model to be synergistically deployed across other projects. IIC will perform assays outlined in the four Specific Aims presented below and conduct bioinformatics and data analysis pipeline to assess the immune alterations induced by the distinct tolerance regimens utilized in the Projects 1, 2 and 3. Aim 1: Define the alterations in the immune landscape of renal transplant recipients by high-dimensional CyTOF profiling of circulating immune cells during induction and maintenance of tolerance. Aim 2: Generate and validate TCR signatures using single cell TCRseq to track donor-antigen specific CD4+ and CD8+ T cells. Aim 3: Define the fate and function of donor-antigen specific CD4+ T cells with MHC class II tetramers and single cell ATAC and RNA sequencing in serial peripheral blood mononuclear cells of tolerant and rejecting renal allograft recipients. Aim 4: Analyze the spatio-temporal organization of immune cells in renal allografts, spleen, and lymph nodes at the transcription level by Digital Spatial Profiling and at the protein level by MIBI-TOF. Analysis of multiple tissue compartments longitudinally to generate a large, but highly controlled, dataset. We intentionally apply this multi-omics approach to advantage parallel, rather than sequential, discrimination of system level analyses of immune mechanisms that contribute to transplant tolerance. Analysis of samples from the 3 projects utilizing distinct tolerance regimens will provide new immunological insights on induction and maintenance of tolerance. Moreover, the performance of assays across projects supports lateral transfer of knowledge to identify commonality, independent of strategy, revealing principal features underlying immune tolerance to advance towards successful clinical application. IIC will interact regularly with the PIs of the 3 Projects and with the administrative core to design, execute, analyze and trouble shoot the studies proposed in this application.