The role of IL-4 and IL-4Ralpha in the multi-step process of Th2 development

IL-4 和 IL-4Ralpha 在 Th2 发育的多步骤过程中的作用

基本信息

批准号：
7725827
负责人：
MARKUS MOHRS
金额：
$ 38.9万
依托单位：
TRUDEAU INSTITUTE, INC.
依托单位国家：
美国
项目类别：
财政年份：
2006
资助国家：
美国
起止时间：
2006-11-15 至 2011-10-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7725827
关键词：
A Mouse Adjuvant Allergic Reaction Animals Antigens Asthma Attention Bone Marrow CD4 Positive T Lymphocytes Cell Lineage Cells Chimera organism Competence Complex Cytokine Receptors Data Development Helminths Human Hypersensitivity Immune response Immunity Immunization In Vitro Infection Interleukin-4 Intervention Larva Mediating Medical Modeling Mouse Strains Mus Nematoda Nematode infections Nematospiroides dubius Parasites Parasitic nematode Pathway interactions Play Population Positioning Attribute Process Production Publications Reaction Reporter Research Design Role Signal Transduction Soil Source T-Lymphocyte Testing Th2 Cells Therapeutic Intervention Transgenic Organisms Work airway hyperresponsiveness atopy base cytokine design gastrointestinal immunopathology in vivo insight interest mouse model novel research study response selective expression

项目摘要

Soil transmitted parasitic nematodes are world wide one of the most commonly acquired infections with multicellular parasites. The adaptive host response in humans and mice is characterized by the presence of CD4+ Th2 cells and their signature cytokine IL-4. Th2 cells and IL-4 are associated with protective immune responses against helminth parasites they are detrimental in certain immunopathologies such as antigen-induced asthmatic reactions, atopy and allergy. While Th2 cells are associated with nematode infections; however, it is not clear how Th2 cells develop from naive CD4+ T cells in response to infection/ Numerous studies have shown that IL-4Ra-mediated signals and IL-4 are required for the Th2 development of naive CD4+ T cells in vitro, however, very little is known about these processes in vivo. This is largely due to the difficulty to identify and track Th2 cells defined by IL-4 expression. To overcome these limitations we have developed bicistronic IL-4 reporter (4get) mice. In contrast to in vitro studies, our preliminary data show that Th2 priming occurs surprisingly efficient in IL4Ra-/- 4get mice infected with the murine helminth H. polygyrus. Thus, the role of IL-4R and IL-4 for Th2 development is controversial. Recently we have generated novel IL-4 dual-reporter mice (4get/KN2) and revealed that IL-4 competence and IL-4 production are distinct steps. Here we propose a novel model whereby Th2 development and IL-4 production occur in several distinct, identifiable steps: 1. activation, 2. differentiation, 3. expansion, 4. IL-4 production, 5. dissemination. In the current application we will revisit the role of IL-4 and IL-4Ra-mediated signals for Th2 differentiation and IL-4production using our unique dual-reporter mouse model. We hypothesize that IL-4R functions are not required to nucleate the Th2 priming of CD4+ T cells but play a role in multiple other steps of this model. In Aim 1 we will analyze which step(s) in Th2 differentiation of the endogenous T cell population are regulated by IL-4R signals in response to infection. In Aim 2 we will determine which step(s) in Th2 differentiation are regulated by IL-4R signals mediated directly on naive, antigen-specific CD4+ T cells. We will adoptively transfer apTCR transgenic CD4+ T cells and immunize with the cognate antigen using H. polygyrus larvae as a Th2 adjuvant. In Aim 3 we will dissect the role of IL-4 and IL-4Ra expression by selective lineages in the multi-step process of Th2 differentiation. We will generate various bone marrow chimeras which lack the respective components in selective cellular lineages and follow the development of the endogenous T helper response to infection. Collectively the proposed Aims will reveal the mechanism by which IL-4R and IL-4 regulate the multiple steps of Th2 development in vivo. These insights are valuable for designing agonistic and antagonistic intervention strategies targeting the IL-4R and IL-4.

土壤传播的寄生线虫是世界范围内最常见的感染之一多细胞寄生虫。人类和小鼠的适应性宿主反应的特点是 CD4+ Th2 细胞及其标志性细胞因子 IL-4 的存在。 Th2 细胞和 IL-4 与针对蠕虫寄生虫的保护性免疫反应，它们在某些免疫病理学中是有害的例如抗原诱发的哮喘反应、特应性和变态反应。虽然 Th2 细胞与线虫感染；然而，尚不清楚 Th2 细胞如何从初始 CD4+ T 细胞发育而来，以响应感染/大量研究表明，Th2 细胞需要 IL-4Ra 介导的信号和 IL-4 幼稚 CD4+ T 细胞在体外发育，然而，人们对体内这些过程知之甚少。这很大程度上是由于难以识别和追踪由 IL-4 表达定义的 Th2 细胞。为了克服这些局限性我们开发了双顺反子 IL-4 报告基因 (4get) 小鼠。与体外研究相比，我们的初步数据表明，Th2 启动在感染了鼠蠕虫 H. polygyrus。因此，IL-4R 和 IL-4 在 Th2 发育中的作用存在争议。最近，我们培育了新型 IL-4 双报告基因小鼠 (4get/KN2)，并揭示了 IL-4 的能力和 IL-4 的生产是不同的步骤。在这里，我们提出了一种新模型，其中 Th2 发育和 IL-4 生产发生在几个不同的、可识别的步骤中：1. 激活，2. 分化，3. 扩增，4. IL-4 生产，5.传播。在当前的应用中，我们将重新审视 IL-4 和 IL-4Ra 介导的作用使用我们独特的双报告基因小鼠模型检测 Th2 分化和 IL-4 产生的信号。我们假设 IL-4R 功能不需要使 CD4+ T 细胞的 Th2 启动成核，但发挥在此模型的多个其他步骤中的作用。在目标 1 中，我们将分析 Th2 分化的哪些步骤内源性 T 细胞群受到 IL-4R 信号的调节以响应感染。在目标 2 中，我们将确定 Th2 分化中的哪些步骤受到直接介导的 IL-4R 信号的调节，抗原特异性 CD4+ T 细胞。我们将过继转移apTCR转基因CD4+ T细胞并进行免疫使用 H.polygyrus 幼虫作为 Th2 佐剂与同源抗原。在目标 3 中，我们将剖析以下角色：在 Th2 分化的多步过程中选择性谱系表达 IL-4 和 IL-4Ra。我们将产生各种骨髓嵌合体，这些嵌合体缺乏选择性细胞谱系中的相应成分并跟踪内源性 T 辅助细胞对感染反应的发展。集体提出的目标将揭示IL-4R和IL-4调节Th2发育多个步骤的机制体内。这些见解对于设计针对目标的对抗性和对抗性干预策略非常有价值 IL-4R和IL-4。