Interplay Between Pathogenic and Regulatory T Cells in EAE

EAE 中致病性 T 细胞和调节性 T 细胞之间的相互作用

基本信息

批准号：
7943400
负责人：
Estelle Bettelli
金额：
$ 31.6万
依托单位：
BENAROYA RESEARCH INST AT VIRGINIA MASON
依托单位国家：
美国
项目类别：
财政年份：
2008
资助国家：
美国
起止时间：
2008-08-01 至 2013-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7943400
关键词：
Accounting Address Affect Allergic Reaction Animal Model Animals Antibodies Antigens Autoimmune Diseases Autoimmune Process Autoimmunity C57BL/6 Mouse CD4 Positive T Lymphocytes CNS autoimmunity Cell physiology Cells Chronic Phase Chronic Phase of Disease Clinical Complement Control Animal Data Development Diphtheria Toxin Disease Disease Progression Encephalomyelitis Experimental Autoimmune Encephalomyelitis Gene Expression Profiling Green Fluorescent Proteins Hand Helper-Inducer T-Lymphocyte IL-23 p19 Immigration Immune response Immune system Immunity Immunization Inflammatory Injection of therapeutic agent Interleukin-12 Interleukin-17 Interleukin-4 Interleukin-6 Intervention Lead Macrophage Activation Mediating Microglia Mouse Strains Multiple Sclerosis Mus Myelin Names Neuraxis Onset of illness Pathogenicity Phase Play Population Positioning Attribute Production RNA Sequences Reaction Reagent Recovery Recruitment Activity Regulation Relative (related person)Reporter Reporting Resistance Role STAT4 gene Series Severity of illness Staging Surface T-Lymphocyte Th1 Cells Th2 Cells Therapeutic Time Transcription factor genes Winged Helix abstracting base cell type cytokine design diphtheria toxin receptor human disease in vivo interleukin-23 knock-down macrophage migration neoplastic cell novel oligodendrocyte-myelin glycoprotein podoplanin public health relevance receptor receptor expression red fluorescent protein research study response selective expression tool transcription factor

项目摘要

DESCRIPTION (provided by applicant): Abstract T helper (h) 17 cells are a newly described subset of CD4+ T cells which has been implicated, along with Th1 cells in the development of autoimmune diseases such as multiple sclerosis (MS) and its animal model experimental autoimmune encephalomyelitis (EAE). Regulatory T cells, on the other hand, protect from the development of autoimmunity. The mechanisms by which Th17 cells and factors associated with their differentiation promote autoimmunity have not been fully elucidated. We have previously determined that the differentiation of Th17 cells required the combined action of TGF-2 plus IL-6 and later IL-21. On the other hand IL-23 p19 is critical for sustaining Th17 responses and for the development of EAE since mice deficient for IL- 23 p19 are resistant to EAE. We found that IL-23 receptor (IL-23R), besides being expressed on Th17 cells, is also expressed on cells of the innate immune system such as macrophages and microglia. However, the role of IL-23 on non-T cells is to date unknown. We have generated a novel IL-23R green fluorescent protein (GFP) knockin (KI) reporter mouse in which all cells expressing the IL-23R will concomitantly express the GFP and become non responsive to IL-23. The IL-23R GFP KI mice will help us determine the relative contribution of IL- 23 on Th17 cells and macrophages/microglia in the development and progression of EAE. We have further determined that Th17 cells migrate quickly to the central nervous system (CNS) at the onset of EAE and their numbers correlate with the disease severity. However, the number of Th17 cells rapidly decline while the number of Th1 cells is maintained suggesting that Th17 cells may have a dominant role early on at the initiation phase and that Th1 cells are more involved during the chronic phase of EAE. We have generated an IL-17F RFP/DTR mouse in which Th17 cells can be deleted at any given time through the injection of diphtheria toxin and we will be able to determine the role of Th17 at different stages of the disease. Finally, in our efforts to further determine the mechanisms by which Th17 cells have enhanced pathogenic activity, we have isolated a surface molecule selectively expressed on Th17 cells named podoplanin. The expression of podoplanin has never been reported on T cells before but podoplanin has previously been implicated in the migration of tumor cells. Our preliminary data, injecting anti-podoplanin antibody in vivo in mice immunized with myelin antigen show an increase number of Th17 cells present in the CNS of the treated animals compared to control animals suggesting that podoplanin may play a role in the migration of Th17 cells. PUBLIC HEALTH RELEVANCE: Based on our preliminary data and using novel reagents that we have generated, we have designed experiments to address several outstanding questions that are important for our understanding of autoimmune diseases: 1) Determine the interplay between effector Th17 and Th1 cells, and T-reg cells during EAE, 2) Determine the role of podoplanin in the development of Th17 cells 3) Determine how IL-23 exacerbate the development of EAE through its action on Th17 and cells of the innate immune system. Project Narrative These studies will complement ongoing studies on Th17 cells and regulatory T cells and contribute to our overall understanding of the mechanisms involved in the development of autoimmunity and their regulation.

描述（由申请人提供）：摘要T助手（H）17细胞是CD4+ T细胞的新描述的子集，与TH1细胞一起在自身免疫性疾病（如多发性硬化症（MS）及其动物模型实验性自身免疫性脑脊镜炎（EAE））中与TH1细胞一起。另一方面，调节性T细胞可以防止自身免疫的发展。 Th17细胞及其分化促进自身免疫性相关的因素的机制尚未得到充分阐明。我们先前已经确定TH17细胞的分化需要TGF-2 Plus IL-6和后来IL-21的合并作用。另一方面，IL-23 p19对于维持Th17响应和EAE的发展至关重要，因为缺乏IL-23 P19的小鼠对EAE具有抗性。我们发现，除了在Th17细胞上表达外，IL-23受体（IL-23R）还在先天免疫系统（例如巨噬细胞和小胶质细胞）的细胞上表达。然而，迄今为止，IL-23对非T细胞的作用是未知的。我们已经产生了一种新型的IL-23R绿色荧光蛋白（GFP）敲击蛋白（Ki）报告基因小鼠，其中所有表达IL-23R的细胞都会同时表达GFP并对IL-23的反应。 IL-23R GFP KI小鼠将帮助我们确定IL-23对Th17细胞和巨噬细胞/小胶质细胞在EAE发育和进展中的相对贡献。我们进一步确定TH17细胞在EAE发作时迅速迁移到中枢神经系统（CNS），其数量与疾病的严重程度相关。然而，TH17细胞的数量迅速下降，而Th1细胞的数量则表明Th17细胞在开始阶段的早期可能具有主要作用，并且在EAE的慢性阶段，Th1细胞更多地参与。我们已经产生了IL-17F RFP/DTR小鼠，其中可以通过注射白喉毒素在任何给定时间删除Th17细胞，我们将能够确定Th17在疾病不同阶段的作用。最后，为了进一步确定Th17细胞增强致病活性的机制，我们已经在名为Podoplanin的Th17细胞上选择性地分离了表面分子。 podoplanin的表达从未在T细胞上报道过，但是Podoplanin以前与肿瘤细胞的迁移有关。我们的初步数据，在用髓磷脂抗原免疫的小鼠体内注射抗杂偶素抗体的体内显示，与对照动物相比，治疗动物中枢神经系统中存在的Th17细胞数量增加，这与对照动物相比表明podoplanin可能在TH17细胞的迁移中起作用。公共卫生相关性：基于我们的初步数据并使用我们生成的新试剂，我们设计了实验，以解决几个重要的问题，这些问题对于我们对自身免疫性疾病的理解很重要：1）确定效应子Th17和Th1细胞之间的相互作用，EAE期间的T-Reg细胞之间的相互作用，以及在EAE期间的T-Reg之间的相互作用，2）确定podoplanin在Th17细胞中的作用3）。它对TH17的作用和先天免疫系统的细胞。项目叙述这些研究将补充对TH17细胞和调节性T细胞的持续研究，并有助于我们对自身免疫发展及其调节所涉及的机制的整体理解。