Reporting IL-17 in Type 1 Diabetes

报告 1 型糖尿病中的 IL-17

基本信息

批准号：
8316091
负责人：
Linda Mac Pherson Bradley
金额：
$ 9.55万
依托单位：
SANFORD BURNHAM PREBYS MEDICAL DISCOVERY INSTITUTE
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-08-09 至 2013-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8316091
关键词：
Acute Address Adoptive Cell Transfers Aftercare Age Alleles Animals Appearance Autoimmune Diseases Autoimmune Responses Backcrossings Bacterial Artificial Chromosomes Biology CD4 Positive T Lymphocytes Cells Chimeric Proteins Complementary DNA Detection Development Diabetes Mellitus Disease Excision Flow Cytometry Future Gene Expression Generations Genes Genetic Genetic Recombination Goals Grant Heterozygote Histology Homozygote Hyperglycemia In Situ Inbred NOD Mice Inflammation Initiator Codon Insulin Insulin-Dependent Diabetes Mellitus Interferon Type II Interleukin-17 Islet Cell Knockout Mice Lead Lymphoid Microinjections Mus Oocytes Pancreas Pathogenesis Pathology Patients Phase Population Production Proteins Recording of previous events Regulation Reporter Reporter Genes Reporting Rest Serum T cell response T-Lymphocyte Technology Testing Th1 Cells Transgenic Mice Transgenic Organisms Translations Universities Withdrawal animal breeding base cell type design embryonic stem cell homologous recombination improved insight insulin dependent diabetes mellitus onset islet mouse model novel novel strategies response restoration tool transcription factor

项目摘要

DESCRIPTION (provided by applicant): Type 1 diabetes (T1D) is currently an incurable disease because the autoimmune response that destroys insulin-producing b islet cells cannot be controlled. Autoreactive CD4 cells are key orchestrators of T1D. Their escalating responses ultimately escape regulation, leading to acute inflammation in the pancreas and diabetes onset. While it is clear that IFN-g producing Th1 cells are essential to disease pathogenesis, whether or when IL-17-producing Th17 cells contribute to T1D is unknown. Th17 cells that secrete IL-17A and to a lesser extent IL-17F have been implicated in several autoimmune diseases. Our studies show that IL-17A- and IFN-g- secreting CD4 T cells are distinct subsets in pancreatic infiltrates during the insulitis phase and at T1D onset in NOD mice, whereas IL-17F is not detected. In addition, elevated serum levels of IL-17A and IFN-g herald the onset of hyperglycemia. We now find that IL-17A+ CD4 T cells greatly accelerate diabetes in normal (WT) NOD mice. On the basis of our findings and the detection of IL-17A+ cells in T1D patients, we hypothesize that Th17 cells are important contributors to the pathogenesis of T1D. However, without additional tools, it will not be possible to directly address mechanisms that regulate the responses of these cells or their effector functions that could be targeted to control the autoimmune response. Our goal is to develop a novel IL-17A reporter mouse on the NOD background that will enable analysis of the development and responses of Th17 cells in situ by lineage tracing during progression to T1D and, in a future R01 grant we plan to pursue studies of the regulation of Th17 cells in T1D. We propose to use bacterial artificial chromosome (BAC)-based gene reporter and gene knockin strategies to create mice that will enable identification and isolation of CD4 cells that acquire Il17a gene expression, as reported by expression of a GFP-Cre fusion protein inserted in the Il17a locus. We will use a double reporter strategy by breeding these animals to Rosa26-YFP NOD mice to indelibly mark cells with a history of IL-17A production. These mice will provide a novel tool to unequivocally study the contributions of Th17 cells to T1D and the potential functional interrelationships between Th1 and Th17 cells that lead to diabetes onset. The ability to identify changes in the development of Th17 cells and their functions will facilitate the design of appropriate strategies to control autoaggressive CD4 cells and provide new insights into the biology of Th17 cells.

描述（申请人提供）：1 型糖尿病（T1D）目前是一种无法治愈的疾病，因为破坏产生胰岛素的 b 胰岛细胞的自身免疫反应无法控制。自身反应性 CD4 细胞是 T1D 的关键协调者。它们不断升级的反应最终逃脱了监管，导致胰腺急性炎症和糖尿病发作。虽然很明显，产生 IFN-g 的 Th1 细胞对于疾病发病机制至关重要，但产生 IL-17 的 Th17 细胞是否或何时促成 T1D 尚不清楚。分泌 IL-17A 和较小程度的 IL-17F 的 Th17 细胞与多种自身免疫性疾病有关。我们的研究表明，在 NOD 小鼠的胰岛炎阶段和 T1D 发作时，分泌 IL-17A 和 IFN-g 的 CD4 T 细胞是胰腺浸润中的不同亚群，而未检测到 IL-17F。此外，IL-17A 和 IFN-g 血清水平升高预示着高血糖的发生。我们现在发现 IL-17A+ CD4 T 细胞极大地加速了正常 (WT) NOD 小鼠的糖尿病发生。根据我们的研究结果和 T1D 患者中 IL-17A+ 细胞的检测，我们假设 Th17 细胞是 T1D 发病机制的重要贡献者。然而，如果没有额外的工具，就不可能直接解决调节这些细胞的反应或其效应器功能的机制，从而控制自身免疫反应。我们的目标是在 NOD 背景下开发一种新型 IL-17A 报告小鼠，通过谱系追踪在 T1D 进展过程中原位分析 Th17 细胞的发育和反应，在未来的 R01 资助中，我们计划继续研究T1D 中 Th17 细胞的调节。我们建议使用基于细菌人工染色体（BAC）的基因报告基因和基因敲入策略来创建小鼠，该小鼠将能够识别和分离获得 Il17a 基因表达的 CD4 细胞，如插入到细胞中的 GFP-Cre 融合蛋白的表达所报告的那样。 Il17a 基因座。我们将使用双报告策略，将这些动物培育为 Rosa26-YFP NOD 小鼠，以不可磨灭地标记具有 IL-17A 产生历史的细胞。这些小鼠将提供一种新的工具来明确研究 Th17 细胞对 T1D 的贡献以及导致糖尿病发作的 Th1 和 Th17 细胞之间潜在的功能相互关系。识别 Th17 细胞发育及其功能变化的能力将有助于设计适当的策略来控制自身攻击性 CD4 细胞，并为 Th17 细胞的生物学提供新的见解。