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（由申请人提供）：1型糖尿病（T1D）当前是一种无法治愈的疾病，因为无法控制损坏产生胰岛素的B胰岛细胞的自身免疫反应。自动反应性CD4细胞是T1D的关键编排者。他们不断升级的反应最终避免了调节，导致胰腺和糖尿病发作的急性炎症。虽然很明显，产生IFN-G的Th1细胞对于疾病发病机理至关重要，但无论是或何时产生IL-17的Th17细胞对T1D造成T1D的贡献都是未知的。分泌IL-17A和较小程度的IL-17F的Th17细胞与几种自身免疫性疾病有关。我们的研究表明，IL-17A和IFN-G-分泌的CD4 T细胞在胰岛炎阶段和NOD小鼠的T1D发作中是胰腺浸润中的不同子集，而未检测到IL-17F。此外，高血糖的发作升高了IL-17A和IFN-G Herald的血清水平。现在，我们发现IL-17A+ CD4 T细胞在正常（WT）NOD小鼠中大大加速糖尿病。根据我们的发现和T1D患者IL-17A+细胞的检测，我们假设Th17细胞是T1D发病机理的重要促进因素。但是，如果没有其他工具，就无法直接解决调节这些单元响应或其效应子函数的机制，这些函数可以针对控制自身免疫反应。我们的目标是在NOD背景上开发一种新型的IL-17A报告基因小鼠，该小鼠将通过在进展为T1D期间进行谱系跟踪来分析Th17细胞原位的发育和反应，并在将来的R01授予中，我们计划在T1D中对Th17细胞调节进行研究。我们建议使用基于细菌的人造染色体（BAC）基因报告基因和基因敲击蛋白策略来创建小鼠，从而可以鉴定和隔离获得IL17A基因表达的CD4细胞，如在IL17A Locus中插入的GFP-Cre-crefusion蛋白的表达所报道。我们将通过将这些动物繁殖到Rosa26-YFP点头小鼠来使用双重记者策略，以不可磨灭地标记IL-17A产生史的细胞。这些小鼠将提供一种新颖的工具，可以明确研究Th17细胞对T1D的贡献以及Th1和Th17细胞之间的潜在功能相互关系，从而导致糖尿病发作。鉴定Th17细胞及其功能变化的变化的能力将有助于设计适当的策略，以控制自构的CD4细胞，并为TH17细胞的生物学提供新的见解。