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

• 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
• 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' 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

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）：尽管已经证明iNOS在针对微生物病原体的宿主防御中起重要作用，但尚未定义INOS在T细胞和慢性炎症性疾病中的确切功能。分泌白介素17（IL-17）和IL-22的Th17细胞包含了与Th1和Th2子集不同的CD4+ T细胞的最近鉴定的子集，并且RORGT已被鉴定为Th17细胞分化的关键转录因子。越来越多的证据表明，TH17免疫反应与各种自身免疫/炎症性疾病的发病机理有关。因此，阻断Th17细胞激活可能会导致发展慢性炎性疾病的新型策略。我们表明，iNOS敲除小鼠表现出更强的Th17细胞分化，而对TH1或TH2细胞谱系的影响没有重大影响。我们证明，在活化的CD4+ T细胞中诱导了iNOS蛋白，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WT和Inos和Inos和Inos的IL-17产生的无供体，SNAP，SNAP，SNAP，剂量依赖性抑制的IL-17产生。另外，硝蛋白蛋白的酪氨酸残基被硝化，导致 RORGT介导的IL-17启动子激活的抑制作用。最后，与对照CD4+CD45RBHI细胞相比，将iNOS - / - CD4+CD45RBHI细胞转移到RAG - / - 小鼠中，与对照组相比，与对照组相比，与对照小鼠相比，IL-17产生细胞的百分比明显高于对照小鼠。结果表明，T细胞衍生的INOS对Th17免疫反应的发展产生，导致炎症的控制。该提议围绕三个目的构成：1）我们将表征通过T细胞衍生的iNOS调节Th17细胞分化的分子机制。 2）我们将表征T细胞衍生的INOS对人Th17，Th1，Th2和Treg细胞发育的功能。 3）我们将分析iNOS在不同细胞室中的作用，包括巨噬细胞，树突状细胞和T细胞在结肠炎的发展中。拟议的研究将定义一种新型的TH17细胞分化的转录抑制剂，并强调T细胞衍生的INOS作为治疗慢性炎性疾病的新型治疗靶标的重要性。