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.

项目摘要 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， 这项申请的三个项目将发挥协同作用， 宽容，这一计划的核心重点。IIC将进行检测，以分析 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。的 移植耐受实验室（TTL）制定了一系列标准操作规程（SOP） 分析NHP移植受者的供体特异性免疫反应，这将在所有国家共享。 协调U19项目中所有项目的分析建议。内核已经优化了 验证，并采用了几种尖端技术和管道，用于生物信息学数据分析， 多个组织隔室的复杂数据集。对来自3个国家的样本进行全面的免疫分析 IIC的项目将有助于避免测定和分析变化，从而协调结果， 将有助于在一个模型中获得的见解在其他项目中协同部署。IIC将执行 以下四个特定目标中概述的试验，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诱导的免疫改变。 目的1：通过高维CyTOF确定肾移植受者免疫景观的改变 在诱导和维持耐受性过程中分析循环免疫细胞。目标2：生成和验证 使用单细胞TCRseq追踪供体抗原特异性CD 4+和CD 8 + T细胞的TCR特征。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CD 4 + T细胞的命运和功能 移植肾耐受和排斥患者外周血单个核细胞的序列分析 受惠人士目的4：分析移植肾、脾和淋巴中免疫细胞的时空组织 通过数字空间轮廓分析在转录水平上和通过MIBI-TOF在蛋白质水平上检测节点。分析 纵向多个组织隔室以产生大的但高度受控的数据集。我们有意 应用这种多组学方法，以利于系统级的并行而不是顺序区分 分析有助于移植耐受的免疫机制。3个项目的样品分析 利用不同的耐受方案将提供新的免疫学见解， 宽容此外，跨项目的检测性能支持知识的横向转移，以确定 共同性，独立于策略，揭示了免疫耐受的主要特征， 走向成功的临床应用。IIC将定期与3个项目的PI以及 管理核心设计，执行，分析和解决本申请中提出的研究问题。