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

项目总结 了解人类1型糖尿病(T1D)的自身免疫反应对于设计成功的 免疫疗法适用于疾病高危人群以及已确诊疾病的人群。在研究的同时 啮齿动物模型在自身免疫过程中给了我们很大的指导，成功的治疗方法的翻译 而在啮齿动物模型到人类临床试验中的预防研究则远没有那么成功。虽然我们知道 关于啮齿动物模型中胰岛浸润T细胞和人类T1D胰岛浸润的大量信息 通过组织学和免疫组织化学，我们以前直接分离的机会有限。 从患有T1D的人中提取淋巴细胞，用于谱系分析和功能能力的研究。我们有 从12个患有T1D的个体获得胰岛分离，并已直接从胰岛中生长或分类，约300 CD4+和CD8+T细胞系或克隆。我们已经证实了16个CD4+T细胞的反应性和对人类白细胞抗原的限制 细胞系或克隆和3个CD8+T细胞系(具有多种反应性)。这项研究的目的 提出全面定义胰岛反应性、人类白细胞抗原限制和广泛表型、RNA 转录组、T细胞受体结构和T细胞受体谱的现有分析，但尚未， 未定性的胰岛浸润性T细胞，重点是胰岛浸润性CD8+T细胞。这些研究将 包括从患有T1D的捐赠者的胰岛中提取的T细胞，我们预计在这一资助期间收到。 胰岛浸润性T细胞将被用作T1D免疫病理研究的工具，更重要的是，对于 全面了解胰岛T细胞在T1D免疫病理中的作用。我们的 假设-胰岛渗入的CD4+和CD8+T细胞将识别广泛的多肽配体， 由胰岛特有的表达和胰岛特有的翻译后修饰/剪接产生，使用 T细胞谱系受限，但表达类似的促炎、记忆表型。这些小岛- 浸润性T细胞将成为现场研究人员研究胰岛浸润性T细胞功能的重要工具/试剂 T1D人源化小鼠模型中的细胞，T细胞受体谱系分析，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分析中，B 细胞、抗原提呈细胞和先天免疫系统。最后，这些研究将向调查人员通报 为T1D高危人群，特别是已确诊的T1D高危人群设计成功的免疫疗法 T1D。这些研究将继续我们作为Hirn的一部分以及与Hirn调查人员的成功合作。

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.