Immunosurveillance and Immunopathology Core

免疫监视和免疫病理学核心

• 批准号: 10622207
• 负责人: Sabarinathan Ramachandran
• 金额: $ 52.87万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-04-20 至 2028-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10622207
• 关键词: ATAC-seq; Adopted; Antigens; Architecture; Bioinformatics; Biological Assay; Blood capillaries; Blood specimen; CD4 Positive T Lymphocytes; CD8-Positive T-Lymphocytes; Cells; Circulation; Clonality; Data Analyses; Data Set; Development; Discrimination; Fingerprint; Genetic Transcription; Heterogeneity; Immune; Immune Tolerance; Immune response; Immunologic Surveillance; Immunologics; Immunosuppression; Immunotherapy; Infiltration; Kidney; Kidney Transplantation; Knowledge; Laboratories; Lateral; Leadership; Locales; MHC Class II Genes; Macaca fascicularis; Maintenance; Metabolic Diseases; Methodology; Mission; Modeling; Molecular Profiling; Organ Transplantation; Performance; Peripheral; Peripheral Blood Mononuclear Cell; Positioning Attribute; Pre-Clinical Model; Procedures; Proliferating; Proteins; Regimen; Regional Anatomy; Regulation; Regulatory T-Lymphocyte; Research Personnel; Resources; Rhesus; Role; Sampling; Series; Sorting; Specificity; Spleen; System; T cell infiltration; T cell receptor repertoire sequencing; T-Lymphocyte; T-cell receptor repertoire; Technology; Testing; Tissues; Transplant Recipients; Transplant-Related Disorder; Transplantation Immunology; Transplantation Tolerance; Tubular formation; Variant; Withdrawal; bioinformatics pipeline; clinical application; complex data; data analysis pipeline; design; digital; exhaustion; experience; high dimensionality; immunopathology; insight; kidney allograft; lymph nodes; multiple omics; nonhuman primate; novel; peripheral blood; programs; spatiotemporal; transcriptome; transcriptome sequencing

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.

项目概要 免疫监视和免疫病理学核心 (IIC)，通过对来自以下组织的样本进行机制研究 该申请的 3 个项目将发挥协同科学作用，重点关注移植 宽容是该计划的核心焦点。 IIC 将进行检测来分析免疫改变 项目 1、2 和 3 中提出的耐受方案诱导的外周循环、移植物和脾脏。 移植耐受实验室 (TTL) 制定了一系列标准操作程序 (SOP) 分析 NHP 移植受者的供者特异性免疫反应，这些反应将在所有 协调 U19 计划中所有项目提出的分析。核心已优化， 验证并采用了多项尖端技术和流程来进行生物信息数据分析 跨多个组织隔室的复杂数据集。对来自 3 个样本的样品进行全面免疫分析 IIC 的项目将有助于避免化验和分析的变化，从而实现结果的统一 将有助于在一个模型中获得的见解，以便在其他项目中协同部署。 IIC将执行 下面提出的四个具体目标中概述的测定并进行生物信息学和数据分析管道 评估项目 1、2 和 3 中使用的不同耐受方案引起的免疫改变。 目标 1：通过高维 CyTOF 定义肾移植受者免疫景观的变化 在诱导和维持耐受过程中循环免疫细胞的分析。目标 2：生成并验证 TCR 签名使用单细胞 TCRseq 追踪供体抗原特异性 CD4+ 和 CD8+ T 细胞。目标 3：定义 具有 MHC II 类四聚体和单细胞 ATAC 的供体抗原特异性 CD4+ T 细胞的命运和功能 耐受和排斥肾同种异体移植物的连续外周血单核细胞的RNA测序 收件人。目标 4：分析同种异体肾移植物、脾脏和淋巴液中免疫细胞的时空组织 通过数字空间分析在转录水平上检测节点，通过 MIBI-TOF 在蛋白质水平上检测节点。分析 纵向多个组织隔室以生成大型但高度控制的数据集。我们故意 应用这种多组学方法来优势并行而不是顺序地区分系统级别 分析有助于移植耐受的免疫机制。 3个项目的样本分析 利用不同的耐受方案将为诱导和维持免疫学提供新的见解 宽容。此外，跨项目的检测性能支持知识的横向转移，以识别 共性，独立于策略，揭示了免疫耐受的主要特征，以促进进步 迈向成功的临床应用。 IIC 将定期与 3 个项目的 PI 以及 设计、执行、分析和解决本申请中提出的研究的管理核心。