T cell-derived iNOS switches off TH17 cell differentiation in inflammation

T 细胞衍生的 iNOS 在炎症中关闭 TH17 细胞分化

基本信息

批准号：
8722431
负责人：
HUABAO XIONG
金额：
$ 42.38万
依托单位：
ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-09-01 至 2017-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8722431
关键词：
Address Affect Asthma Autoimmune Process Autoimmune Responses Binding CD4 Positive T Lymphocytes Cell Culture Techniques Cell Differentiation process Cell Lineage Cells Chronic Colitis Crohn&apos s disease Dendritic Cells Development Disease Dose Family Feedback Gene Activation Gene Expression Genes Goals Helper-Inducer T-Lymphocyte Host Defense Human IRF4 gene Immune Immune response In Vitro Infiltration Inflammation Inflammatory Inflammatory Bowel Diseases Interleukin-10 Interleukin-17 Interleukin-4 Interleukin-6 Interleukins Knockout Mice Lead Light Mediating Molecular Multiple Sclerosis Mus Mutant Strains Mice Nitrates Nitric Oxide Nitric Oxide Donors Nuclear Orphan Receptor Pathogenesis Play Population Principal Investigator Production Promoter Regions Proteins Regulation Regulatory T-Lymphocyte Reporting Research Rheumatoid Arthritis Role S-nitro-N-acetylpenicillamine STAT3 gene Structure System T cell differentiation T-Lymphocyte Th2 Cells Tissues Transforming Growth Factors Tyrosine Ulcerative Colitis cytokine human NOS2A protein in vivo inhibitor/antagonist interleukin-21 interleukin-22 interleukin-23 mRNA Expression macrophage member microbial new therapeutic target nitration novel novel strategies novel therapeutic intervention pathogen programs promoter public health relevance reconstitution therapeutic target transcription factor

项目摘要

DESCRIPTION (provided by applicant): Although it has been demonstrated that iNOS plays an important role in host defense against microbial pathogens, the exact function of iNOS in T cells and chronic inflammatory diseases has not been defined. TH17 cells, which secrete interleukin 17 (IL-17) and IL-22 comprise a recently identified subset of CD4+ T cells distinct from the TH1 and TH2 subsets and RORgt has been identified as a key transcription factor for TH17 cell differentiation. Increasing evidence indicates that TH17 immune responses are involved in the pathogenesis of various autoimmune/inflammatory diseases. Thus, blocking TH17 cell activation could lead to the development of novel strategies for the treatment of chronic inflammatory diseases. We show that iNOS knock out mice display more robust TH17 cell differentiation without major effects on either TH1 or TH2 cell lineages. We demonstrated that iNOS protein was induced in activated CD4+ T cells and the use of an iNOS selective inhibitor L-NIL significantly increased the percentage of IL-17-producing CD4+ T cells in WT cell cultures, while, an NO donor, SNAP, dose-dependently suppressed IL-17 production in WT and iNOS-/- T cell cultures. In addition, tyrosine residues of RORgt protein were nitrated resulting in the inhibition of RORgt-mediated IL-17 promoter activation. Finally, transfer of iNOS-/- CD4+CD45Rbhi cells into RAG-/- mice induces more severe colitis compared to control CD4+CD45Rbhi cells and mice reconstituted with iNOS-/- cells had a significantly higher percentage of IL-17-producing cells than control mice. The results suggest that T cell-derived iNOS negatively regulates the development of TH17 immune responses resulting in the control of inflammation. This proposal is structured around three aims: 1) We will characterize the molecular mechanisms involved in the regulation of TH17 cell differentiation by T cell-derived iNOS. 2) We will characterize the function of T cell- derived-iNOS on human TH17, TH1, TH2, and Treg cell development. 3) We will analyze the roles of iNOS in different cell compartments including macrophages, dendritic cells, and T cells in the development of colitis. The proposed studies will define a novel transcriptional inhibitor of TH17 cell differentiation and highlight th importance of T cell-derived iNOS as a novel therapeutic target for the treatment of chronic inflammatory diseases.

描述（由申请人提供）：尽管已证明iNOS在宿主防御微生物病原体方面发挥重要作用，但iNOS在T细胞和慢性炎症性疾病中的确切功能尚未确定。 TH17 细胞分泌白细胞介素 17 (IL-17) 和 IL-22，包含最近鉴定的 CD4+ T 细胞亚群，与 TH1 和 TH2 亚群不同，RORgt 已被确定为 TH17 细胞分化的关键转录因子。越来越多的证据表明 TH17 免疫反应参与各种自身免疫/炎症性疾病的发病机制。因此，阻断 TH17 细胞激活可能会导致开发治疗慢性炎症性疾病的新策略。我们发现 iNOS 敲除小鼠表现出更强大的 TH17 细胞分化，且对 TH1 或 TH2 细胞谱系没有重大影响。我们证明 iNOS 蛋白在活化的 CD4+ T 细胞中被诱导，并且使用 iNOS 选择性抑制剂 L-NIL 显着增加了 WT 细胞培养物中产生 IL-17 的 CD4+ T 细胞的百分比，而 NO 供体 SNAP 剂量-在WT和iNOS-/- T细胞培养物中依赖性抑制IL-17的产生。此外，RORgt 蛋白的酪氨酸残基被硝化，导致抑制 RORgt 介导的 IL-17 启动子激活。最后，与对照 CD4+CD45Rbhi 细胞相比，将 iNOS-/- CD4+CD45Rbhi 细胞转移到 RAG-/- 小鼠中会诱导更严重的结肠炎，并且用 iNOS-/- 细胞重建的小鼠具有显着更高的 IL-17 产生细胞百分比比对照小鼠。结果表明，T 细胞衍生的 iNOS 负向调节 TH17 免疫反应的发展，从而控制炎症。该提案围绕三个目标构建：1）我们将描述 T 细胞衍生的 iNOS 调节 TH17 细胞分化所涉及的分子机制。 2) 我们将描述 T 细胞衍生的 iNOS 对人类 TH17、TH1、TH2 和 Treg 细胞发育的功能。 3）我们将分析iNOS在不同细胞区室（包括巨噬细胞、树突状细胞和T细胞）在结肠炎发展中的作用。拟议的研究将定义一种新型 TH17 细胞分化转录抑制剂，并强调 T 细胞衍生的 iNOS 作为治疗慢性炎症性疾病的新型治疗靶点的重要性。