Human Islet-Infiltrating T Cell Biology: Reactivity, Structure, and Function

人类胰岛浸润 T 细胞生物学：反应性、结构和功能

• 批准号: 9459661
• 负责人: David Marshall Harlan
• 金额: $ 197.23万
• 依托单位: UNIV OF MASSACHUSETTS MED SCH WORCESTER
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-09-15 至 2021-08-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9459661
• 关键词: Activities of Daily Living; Affinity; Antigen-Presenting Cells; Antigens; Autoantigens; Autoimmune Process; Autoimmune Responses; B-Lymphocytes; Binding; Biological Assay; Biological Preservation; Blocking Antibodies; CD4 Positive T Lymphocytes; CD8-Positive T-Lymphocytes; CD8B1 gene; Cell Communication; Cell Line; Cells; Cellular biology; Childhood; Clinical Trials; Clone Cells; Collaborations; Complex; Coupled; Crystallization; Diabetes Mellitus; Diabetes autoantibodies; Discrimination; Disease; Engineering; Epitopes; Funding; Future; Gene Expression; Histology; Home environment; Human; Immunohistochemistry; Immunotherapy; In Situ; Inbred NOD Mice; Individual; Infiltration; Innate Immune System; Insulin; Insulin-Dependent Diabetes Mellitus; Islet Cell; Islets of Langerhans; Kinetics; Left; Ligands; Lymphocytic Infiltrate; Memory; Methods; Modeling; Modification; Molecular; Molecular Cloning; Molecular Profiling; Organ Donor; Pancreas; Pathogenesis; Pathogenicity; Peptide/MHC Complex; Peptides; Peripheral; Phenotype; Post-Translational Protein Processing; Prevention; Proteins; RNA; RNA Splicing; Reagent; Receptor Cell; Recovery; Regulatory T-Lymphocyte; Reporting; Research Design; Research Personnel; Risk; Rodent Model; Sampling; Specificity; Spleen; Structure; T-Cell Receptor; T-Lymphocyte; Techniques; Translations; United States; Universities; alpha-beta T-Cell Receptor; autoreactive T cell; autoreactivity; base; cell type; clinical Diagnosis; cytokine; cytotoxicity; design; disorder risk; experimental study; human disease; human subject; humanized mouse; immunopathology; insight; insulin dependent diabetes mellitus onset; islet; lymph nodes; mouse model; peripheral blood; programs; success; tool; transcriptome; transcriptome sequencing

PROJECT SUMMARY Understanding the autoimmune response in human type 1 diabetes (T1D) is crucial for the design of successful immunotherapies for those at-risk for the disease as well as for those with established disease. While studies of rodent models of have greatly instructed us on the autoimmune process, the translation of successful therapies and prevention studies in the rodent models to human clinical trials has been far less successful. While we know much information concerning islet infiltrating T cells in the rodent models and islet infiltrates in humans with T1D by histology and immunohistochemistry, we previously have had limited opportunity to isolate directly the lymphocytic infiltrate from humans with T1D for studies of repertoire analysis and functional capacity. We have received islet isolations from 12 individuals with T1D and have grown or sorted directly out of the islets, ~300 CD4+ and CD8+ T cell lines or clones. We have demonstrated the reactivity and HLA restriction of 16 CD4+ T cell lines or clones and 3 CD8+ T cells lines (multiple reactivities are represented). The purpose of the study proposed here is to comprehensively define the islet reactivity, HLA restriction and extensive phenotype, RNA transcriptome, T cell receptor structure and T cell receptor repertoire analysis of the existing, but yet, uncharacterized islet infiltrating T cells, with an emphasis on islet-infiltrating CD8+ T cells. These studies will include T cells derived from the islets of donors with T1D which we anticipate receiving during this funding period. Islet-infiltrating T cells will be used as tools for the immunopathologic study of T1D and importantly, for comprehensive understanding of the function of islet-infiltrating T cells in T1D immunopathology. Our hypothesis is that-islet infiltrating CD4+ and CD8+ T cells will recognize a broad range of peptide ligands, resulting from both islet-specific expression and islet-specific post-translational modification/splicing, using restricted T cell repertoires, but expressing a similar proinflammatory, memory phenotypes. These islet- infiltrating T cells will be vital tools/reagents for investigators in the field to study the function of islet-infiltrating T cells in humanized mouse models of T1D, T cell receptor repertoire analysis, for preservation of autoreactive T cell receptors for future studies, in functional assays of autoreactive T cell interactions with T regulatory cells, B cell, antigen presenting cells and the innate immune system. Finally, these studies will inform investigators in the design of successful immunotherapies for those at-risk for T1D and especially for those with established T1D. The studies will continue our successful collaborations as part of HIRN and with HIRN investigators.

项目摘要 了解人类1型糖尿病（T1 D）的自身免疫反应对于设计成功的治疗方案至关重要。 免疫疗法对于那些处于疾病风险中的人以及对于那些已经确定的疾病。虽然研究 啮齿类动物模型在自身免疫过程，成功疗法的转化， 从啮齿动物模型到人类临床试验的预防研究远没有那么成功。虽然我们知道 关于啮齿动物模型中胰岛浸润T细胞和T1 D患者胰岛浸润的大量信息 通过组织学和免疫组织化学，我们以前只有有限的机会直接分离出 来自患有T1 D的人的淋巴细胞浸润用于研究库分析和功能能力。我们有 从12名T1 D患者中分离胰岛，并直接从胰岛中生长或分选，约300 CD 4+和CD 8 + T细胞系或克隆。我们已经证明了16个CD 4 + T细胞的反应性和HLA限制性， 细胞系或克隆和3个CD 8 + T细胞系（代表多种反应性）。研究的目的 本文提出的是全面定义胰岛反应性、HLA限制性和广泛表型、RNA 转录组、T细胞受体结构和T细胞受体库分析的现有研究，但至今， 未表征的胰岛浸润T细胞，重点是胰岛浸润CD 8 + T细胞。这些研究将 包括来自T1 D捐赠者胰岛的T细胞，我们预计在此资助期间将收到这些细胞。 胰岛浸润T细胞将被用作T1 D免疫病理学研究的工具，重要的是， 全面了解胰岛浸润T细胞在T1 D免疫病理学中的功能。我们 假设是胰岛浸润CD 4+和CD 8 + T细胞将识别广泛的肽配体， 由胰岛特异性表达和胰岛特异性翻译后修饰/剪接产生，使用 限制性T细胞库，但表达类似的促炎性记忆表型。这些小岛- 浸润性T细胞将成为该领域研究人员研究胰岛浸润性T细胞功能的重要工具/试剂 T1 D的人源化小鼠模型中的T细胞，T细胞受体库分析，用于保存自身反应性T细胞 用于未来研究的细胞受体，在自身反应性T细胞与T调节细胞相互作用的功能测定中，B 细胞、抗原呈递细胞和先天免疫系统。最后，这些研究将为研究人员提供信息， 为那些有T1 D风险的人设计成功的免疫疗法，特别是那些已经确定的 T1 D。这些研究将继续我们作为HIRN的一部分与HIRN研究人员的成功合作。