TOLLIP and Tuberculosis Immunopathogenesis

TOLLIP 和结核病免疫发病机制

• 批准号: 8721841
• 负责人: Javeed Ali Shah
• 金额: $ 17.71万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-08-15 至 2018-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8721841
• 关键词: Address; Adjuvant; Adult; Affect; Anti-Inflammatory Agents; Anti-inflammatory; Appointment; Autophagocytosis; BCG Vaccine; Binding; Biological; Biological Assay; Biology; Blood specimen; Case-Control Studies; Caucasians; Caucasoid Race; Cell Maturation; Cell membrane; Cell physiology; Cells; Cellular Immunology; Cellular biology; Chicago; Childhood; Clinical; Clinical Research; Cohort Studies; Communicable Diseases; Data; Dendritic Cells; Development; Endocytosis; Environment; Faculty; Fellowship; Funding; Genetic; Genetic Markers; Genetic Polymorphism; Genetic Variation; Goals; Grant; Health; Host Defense; Human; Human Genetics; Hypersensitivity; Immune; Immune response; Immune system; Immunologics; Immunology; In Vitro; Individual; Infant; Infection; Infection Control; Inflammatory Response; Interleukin-1; Interleukin-6; International; Knowledge; Laboratories; Lead; Ligands; Link; Maps; Mediating; Medicine; Membrane Protein Traffic; Mentors; Mentorship; Metric; Molecular Biology; Morbidity - disease rate; Mus; Mycobacterium tuberculosis; Natural Immunity; Nature; Pathogenesis; Patients; Phagocytosis; Pharmaceutical Preparations; Phospholipids; Physicians; Play; Population; Population Study; Positioning Attribute; Predisposition; Production; Protein Biochemistry; Protein Deficiency; Publications; Receptor Signaling; Regulation; Research; Resources; Risk; Role; Running; Scientist; Shapes; Signal Pathway; Signal Transduction; Source; South Africa; Specialist; Stimulus; T cell response; T-Lymphocyte; TLR2 gene; TLR4 gene; TNF gene; Techniques; Time; Toll-Like Receptor Pathway; Toll-like receptors; Training; Training Programs; Tuberculosis; Tuberculosis Vaccines; Umbilical Cord Blood; United States National Institutes of Health; Universities; Vaccinated; Vaccine Adjuvant; Vaccines; Variant; Vietnam; Washington; Work; base; case control; cohort; cytokine; design; experience; human TOLLIP protein; human population study; immune function; improved; in vivo; insight; interest; mRNA Expression; macrophage; medical schools; meetings; member; monocyte; mortality; mycobacterial; novel vaccines; pathogen; peripheral blood; protein function; protein transport; receptor; research study; response; small hairpin RNA; therapeutic vaccine; tuberculosis immunity; uptake; vaccination against tuberculosis

DESCRIPTION (provided by applicant): The applicant is a specialist in infectious diseases with a long-term interest in tuberculosis and international health. His background in cellular immunology is the basis for the proposed research. He studied cellular immunology of T cells and dendritic cells at the NIH as Howard Hughes Research Scholar as well as protein biochemistry as an undergraduate at the University of Chicago. He has an excellent publication record from his previous research experiences at the NIH, in the laboratory of Robert Seder and from work performed in the laboratory of Thomas Hawn. His immediate goals are to study the mechanisms of TOLLIP regulation of the innate immune response. He will accomplish this using a number of specialized resources as part of a training program. First, his mentor (Dr. Thomas Hawn) has experience studying common genetic variation in large human cohorts that will be applied for the first time to TOLLIP. Second, his co-mentor (Dr. Alan Aderem) has extensive experience in macrophage biology and will allow the applicant to study the role of TOLLIP variation on phagocytosis and autophagy. Third, he will determine the breadth of the effects of TOLLIP on the immune system by studying how TOLLIP may affect dendritic cell maturation, T cell polarization, and cytokine responses. The applicant's long term goals are to seek a clinical appointment in a Division of Infectious Diseases at an academic center, develop sources of independent research funding, and devote >75% of his time to running a lab and collaborative clinical studies. Tuberculosis stimulates intense inflammatory response, but regulation of this response is key to control of infection. The regulation and nature of an optimal inflammatory response to Mycobacterium tuberculosis (MTb) remains poorly understood in humans. Insight into mechanisms of negative regulation of the TLR- mediated innate immune response to MTb could provide significant breakthroughs in the design of new vaccines and drugs. We hypothesized that TOLLIP and its common variants negatively regulate TLR signaling in human monocytes and are associated with susceptibility to tuberculosis. Using shRNA knockdown of TOLLIP in peripheral blood human monocytes, we found that TOLLIP suppresses TNF and IL-6 production after stimulation with TLR2 and TLR4 ligands. We also discovered a common polymorphism (rs5743899) that is associated with TOLLIP deficiency in a Caucasian population. Furthermore, in a case-population study in Vietnam with 760 cord-blood samples and 671 TB case patients, we found that the SNP rs5743899 was associated with susceptibility to tuberculosis (p=6.97x10-7). Together, these data demonstrate that TOLLIP has an anti-inflammatory effect on TLR signaling in humans and that TOLLIP deficiency is associated with an increased risk of TB. These data implicate an unexpected mechanism of negative regulation of TLR signaling in human TB pathogenesis. In this application, we will examine human variants of TOLLIP and their role in regulating macrophage, dendritic cell, and T cell responses to MTb infection. In Aim 1, I will determine the specific polymorphisms commonly found in the population that affect TOLLIP function and influence risk for developing tuberculosis. Furthermore, we will more fully characterize how TOLLIP influences phagocytosis, autophagy, and mycobacterial replication in macrophages and dendritic cells. I will also combine our human population studies with mechanistic studies in Tollip -/- mice. In Aim 2, I will use multiple techniques, including mouse studies, study of ex vivo infant cytokine response to BCG, and a unique case-control study of pediatric TB in South Africa to study the in vitro and in vivo effects of TOLLIP variation on dendritic cell biology and the effect of TOLLIP variation on the regulation of innate immune priming of the adaptive immune response. This work will be performed at the University of Washington School of Medicine in Seattle. The applicant and proposed mentor are in the Division of Allergy and Infectious Diseases, which consists of 73 fulltime faculty members whose total grant support exceeds $135 million annually. This Division is an ideal environment for training physician-scientists as more than 80% of past fellowship trainees have obtained faculty positions in academic medicine. The applicant proposed research takes advantage of his strong background in cellular immunology but requires additional training in molecular biology, innate immunology, and human genetics. He has detailed coursework, seminars, and meetings to address these needs and has developed metrics for assessing his progress. He has also assembled a scientific advisory panel to provide one-on-one expertise. Finally, the proposed mentor has a demonstrated track record with these techniques and has successfully mentored several other junior scientists. NARRATIVE: Understanding the immunologic and genetic factors that confer protection from tuberculosis is vital to the development of successful vaccines and therapeutics. TOLLIP is a recently discovered regulator of the Toll-like receptor pathway that is critical for an effective immune response to tuberculosis. I propose to focus on the mechanisms by which TOLLIP regulates innate immunity by determining the specific common variants that affect TOLLIP function. I will also examine the role TOLLIP plays in regulating cellular uptake and destruction of tuberculosis. Lastly, I will study how TOLLIP influences long term immune responses to BCG in infants. I will correlate this work with ongoing studies in Tollip-deficient mice This work will provide important information about the role of TOLLIP in influencing the immune response to tuberculosis, provide insight into the components of an effective immune response to infections, and may lead to improved vaccines and vaccine adjuvants to tuberculosis.

描述（由申请人提供）：申请人是传染病专家，对结核病和国际卫生有长期兴趣。他在细胞免疫学方面的背景是拟议研究的基础。他作为霍华德休斯研究学者在美国国立卫生研究院研究 T 细胞和树突状细胞的细胞免疫学，并在芝加哥大学读本科时研究蛋白质生物化学。他之前在 NIH、Robert Seder 实验室以及 Thomas Hawn 实验室进行的研究经历使他拥有出色的发表记录。他的近期目标是研究 TOLLIP 对先天免疫反应的调节机制。作为培训计划的一部分，他将使用许多专业资源来实现这一目标。首先，他的导师（Thomas Hawn 博士）拥有研究大型人类群体中常见遗传变异的经验，这些经验将首次应用于 TOLLIP。其次，他的共同导师（Alan Aderem 博士）在巨噬细胞生物学方面拥有丰富的经验，将允许申请人研究 TOLLIP 变异对吞噬作用和自噬的作用。第三，他将通过研究 TOLLIP 如何影响树突状细胞成熟、T 细胞极化和细胞因子反应，确定 TOLLIP 对免疫系统影响的广度。申请人的长期目标是在学术中心的传染病科寻求临床任命，开发独立研究资金来源，并将超过 75% 的时间用于运行实验室和合作临床研究。结核病会刺激强烈的炎症反应，但调节这种反应是控制感染的关键。人类对结核分枝杆菌 (MTb) 最佳炎症反应的调节和性质仍然知之甚少。深入了解 TLR 介导的针对 MTb 的先天免疫反应的负调节机制可以为新疫苗和药物的设计提供重大突破。我们假设 TOLLIP 及其常见变体负向调节人类单核细胞中的 TLR 信号传导，并与结核病易感性相关。通过对人外周血单核细胞中 TOLLIP 的 shRNA 敲除，我们发现 TOLLIP 在 TLR2 和 TLR4 配体刺激后抑制 TNF 和 IL-6 的产生。我们还发现了一种常见的多态性 (rs5743899)，该多态性与白种人群体中 TOLLIP 缺陷相关。此外，在越南开展的一项包含 760 份脐带血样本和 671 名结核病患者的病例群体研究中，我们发现 SNP rs5743899 与结核病易感性相关 (p=6.97x10-7)。总之，这些数据表明 TOLLIP 对人类 TLR 信号传导具有抗炎作用，并且 TOLLIP 缺乏与结核病风险增加相关。这些数据暗示了人类结核病发病机制中 TLR 信号传导负调控的意外机制。在此应用中，我们将研究 TOLLIP 的人类变体及其在调节巨噬细胞、树突状细胞和 T 细胞对 MTb 感染的反应中的作用。在目标 1 中，我将确定人群中常见的影响 TOLLIP 功能并影响患结核病风险的特定多态性。此外，我们将更全面地描述 TOLLIP 如何影响巨噬细胞和树突细胞中的吞噬作用、自噬和分枝杆菌复制。我还将把我们的人群研究与 Tollip -/- 小鼠的机制研究结合起来。在目标 2 中，我将使用多种技术，包括小鼠研究、体外婴儿细胞因子对卡介苗反应的研究，以及南非儿童结核病的独特病例对照研究，以研究体外和体内效应 TOLLIP 变异对树突状细胞生物学的影响以及 TOLLIP 变异对适应性免疫反应的先天免疫启动调节的影响。这项工作将在西雅图华盛顿大学医学院进行。申请人和拟议的导师来自过敏和传染病部门，该部门由 73 名全职教员组成，每年的资助总额超过 1.35 亿美元。该部门是培训医师科学家的理想环境，因为超过 80% 的过去的研究员学员获得了学术医学教职。申请人提出的研究利用了他在细胞免疫学方面的强大背景，但需要在分子生物学、先天免疫学和人类遗传学方面接受额外的培训。他通过详细的课程作业、研讨会和会议来满足这些需求，并制定了评估进度的指标。他还组建了一个科学顾问小组来提供一对一的专业知识。最后，拟议的导师在这些技术方面拥有良好的记录，并成功指导了其他几位初级科学家。 叙述：了解预防结核病的免疫和遗传因素对于成功开发疫苗和治疗方法至关重要。 TOLLIP 是最近发现的 Toll 样受体途径的调节剂，该途径对于结核病的有效免疫反应至关重要。我建议通过确定影响 TOLLIP 功能的特定常见变异来重点关注 TOLLIP 调节先天免疫的机制。我还将研究 TOLLIP 在调节细胞摄取和破坏结核病方面发挥的作用。最后，我将研究 TOLLIP 如何影响婴儿对 BCG 的长期免疫反应。我将把这项工作与正在进行的 Tollip 缺陷小鼠研究联系起来这项工作将 提供有关 TOLLIP 在影响结核病免疫反应中的作用的重要信息，深入了解对感染的有效免疫反应的组成部分，并可能导致改进结核病疫苗和疫苗佐剂。