Dys/Regulation of the Immune System in Autoimmunity

描述(申请人提供)：本课程的目标是了解自身免疫中免疫系统的调节和失调。该计划涉及三个部门成员之间的协作互动，并由三个核心设施支持的四个项目组成。免疫学、分子生物学和生物化学领域的专业知识将侧重于启动和维持自身免疫反应的事件，以及包含自身免疫的调节过程。我们将解决以下问题。启动自身免疫反应的要求是什么？自身免疫反应受调控吗？如果是的话，是通过什么机制调控的？在正常情况下，免疫调节是否包含自身免疫反应？最后，持续的自身免疫反应是否仍然是慢性的，因为它们不同于单一的启动反应，而不是对来自同一组织的其他自身抗原的反应？这些问题将通过这些项目的首席调查员之间的协作互动来解决，这些互动如下： (1)R.A.Flavell-Dr.Flavell将阐述IL-10在自身免疫性疾病中的作用。首先，他将开发一种新的模型系统，以可视化体外和体内IL-10的产生。使用GFP敲入策略，可以通过GFP表达检测到产生IL-10的细胞。这将被用来可视化EAE和IBD期间IL-10的产生，并可用于监测一些干预策略对IL-10的影响，包括Janeway博士使用皮肤贴片进行的“治疗性疫苗接种”。他还将通过转基因阻断IL-10作用的受体的小鼠，研究IL-10是否作用于T细胞或抗原提呈细胞，以介导其对EAE和IBD的影响。此前，他曾成功地将这一方法用于另一种调节细胞因子转化生长因子-β。因此，这种方法似乎很有希望。 (2)小C.A.Janeway-这个项目将检验在B10.PL背景下转基因MBP特异性克隆的TCR的小鼠产生抑制性或调节性T细胞的能力。他们最近观察到，在诱发和自发疾病中，它们都可以产生这样的细胞。他们将确定他们是否能在正常的B10.PI小鼠和对其他自身抗原有反应的小鼠中观察到相同的表型。 (3)M.J.Shlomchik转基因小鼠模型将用于研究B细胞在正常小鼠和自身免疫小鼠中表达疾病相关自身抗体类风湿因子(RF)的调节。这些B细胞在自身免疫倾向的小鼠中自发激活，而在正常小鼠中保持静止。这一系统将用于：1)确定疾病的发生和早期传播所需的环境和遗传因素；2)确定从最初的自身反应性B细胞增殖的结节到慢性持续疾病的一连串事件；以及3)在表型和分子水平上确定逃脱外周耐受的独特B细胞系的身份。 (4)M.J.Mamula--总的来说，这项建议将研究系统性自身免疫性疾病中B细胞和T细胞的相互作用。这项工作将利用自身抗体转基因小鼠来确定它们在维持正常动物品系的免疫耐受方面的作用，以及在自身免疫倾向小鼠的诱导自身免疫和病理方面的作用。其他研究将检验体内自身抗原的水平和暴露如何控制B和T细胞自身免疫的诱导和持续。 这四个项目将得到一个行政核心(核心A)的支持，以协调整个项目；一个转基因小鼠核心(核心B)，以提供大多数这些研究所必需的基因靶向和转基因啮齿动物；以及一个细胞分离核心(核心C)，使我们能够分离和分析所有这些项目中的细胞。该计划由计划教职员工和研究人员的频繁会议协调，将这些不同的方法聚集在一起，以解决共同的目标。

DESCRIPTION (provided by applicant): The goal of this program is to understand the regulation and disregulation of the immune system in autoimmunity. The program involves collaborative interactions between members of three departments, and is organized into four projects supported by three core facilities. Expertise in the field of immunology, molecular biology, and biochemistry will focus on the events that initiate and sustain autoimmune responses, and the regulatory processes, which contain autoimmunity. We will address the following questions. What are the requirements to initiate autoimmune responses? Are autoimmune responses regulated, and if so, by what mechanisms? Does immune regulation contain autoimmune responses under normal circumstances? Finally, do sustained autoimmune responses remain chronic because they diversify from a single initiating response to responses to other autoantigens from the same tissue? These questions will be addressed by collaborative interactions between the Principal Investigators of these projects, which are as follows: (1) R.A. Flavell -Dr. Flavell will address the role of IL-10 in autoimmune disease. First, he will develop a novel model system to visualize IL-10 production in vitro and in vivo. Using a GFP knock-in strategy, IL-10 producing cells will be detectable through GFP expression. This will be used to visualize IL-10 production during EAE and IBD and can be used to monitor the effects on IL-10 of a number of interventional strategies, including Dr. Janeway's "therapeutic vaccination" using skin patch. He will also address whether IL-10 acts on T cells or antigen presenting cells in mediating its effects on EAE and IBD using mice transgenic for receptors that block IL-10 action. He has previously used this approach successfully for the other regulatory cytokine TGF-beta. This approach therefore appears promising. (2) C.A. Janeway Jr. -This project will examine the ability to generate suppressor or regulatory T cells in mice that are transgenic for the TCR of the MBP specific clone on a B10.PL background. They have recently observed that they can generate such cells in both induced and spontaneous disease. They will determine if they can observe the same phenotype in normal B10.PI mice, and mice reactive to other autoantigens. (3) M.J. Shlomchik -Transgenic mouse models will be used to study the regulation of B cells expressing a disease- related autoantibody, rheumatoid factor (RF), in normal and autoimmune mice. These B cells are activated spontaneously in autoimmune-prone mice, yet remain quiescent in normal mice. This system will be used to: 1 ) determine the factors -environmental and genetic -that are required for the onset and early propagation of disease; 2) define the cascade of events that leads from an initial nidus of proliferating autoreactive B cells to chronic, ongoing disease; and 3) determine the identities, at the phenotypic and molecular level, of the unique B lineage cells that have escaped peripheral tolerance. (4) M.J. Mamula -Overall, this proposal will examine B cell- T cell interactions in systemic autoimmune disease. The work will make use of autoantibody transgenic mice to determine their role in the maintenance of immune tolerance in normal strains of animals versus the induction of autoimmunity and pathology in autoimmune prone mice. Additional studies will examine how the levels and exposure to self antigens in vivo control the induction and perpetuation of B and T cell autoimmunity. These four projects will be supported by an Administrative Core (Core A) to coordinate the project as a whole, a genetically modified mouse core (Core B) to provide gene targeted and transgenic rodents essential to most of these studies, and a Cell Separation Core (Core C) to allow us to isolate cells and to analyze cells in all of these projects. The program is coordinated by frequent meetings of the program faculty and research workers bringing together these diverse approaches to address a common goal.