IL-2 Family Cytokines and their Receptors-- Biology of the IL-2 system

IL-2 家族细胞因子及其受体——IL-2 系统的生物学

基本信息

批准号：
8746595
负责人：
Warren J Leonard
金额：
$ 130.4万
依托单位：
NATIONAL HEART, LUNG, AND BLOOD INSTITUTE
依托单位国家：
美国
项目类别：
财政年份：
资助国家：
美国
起止时间：
至
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/8746595
关键词：
Adoptive Immunotherapy Allergic Animals Autoimmunity Binding Biological Biological Process Biology Boxing CD27 Antigens Cell Death Cell Differentiation process Cell physiology Cells ChIP-seq Clinical Collaborations Complex Coupled Cytokine Receptors Cytokine Signaling DNA Sequence Analysis Development Disease Event Family Gene Expression Generations Genes Goals Helminths Hematological Disease Host Defense Human Hypersensitivity IL7R gene Immune Immune response Immune system Immunologic Deficiency Syndromes Inflammatory Inflammatory Bowel Diseases Interleukin 2 Receptor Gamma Interleukin-15 Interleukin-17 Interleukin-2 Interleukin-4 Interleukin-7 Interleukin-9 JAK3 gene Learning MAPK Signaling Pathway Pathway Malignant Neoplasms Mediating Molecular Molecular Profiling Mus Mutate Mutation Phenotype Phosphoric Monoester Hydrolases Phosphotransferases Physiological Process Property Proteins Psoriasis Regulation Relative (related person)Reporter Reporting Retina Role STAT5A gene STAT5B gene Signal Transduction Specificity System T-Cell Receptor T-Lymphocyte T-bet protein Th2 Cells Time Transgenic Mice Transgenic Model Variant Virus Work X-Linked Severe Combined Immunodeficiency autoimmune uveitis base chromatin immunoprecipitation cytokine genome-wide human disease improved in vivo insight interleukin-12 receptor mouse model neoplastic novel pathogen receptor response three dimensional structure

项目摘要

The IL-2 receptor and related cytokine receptor systems are being studied to clarify the T cell immune response in normal, neoplastic, and immunodeficient states. Following T-cell activation by antigen, the magnitude and duration of the T-cell immune response is determined by the amount of IL-2 produced, levels of receptors expressed, and time course of each event. The IL-2 receptor contains three chains, IL-2Ra, IL-2Rb, and gc. Dr. Leonard cloned IL-2Ra in 1984, we discovered IL-2Rb in 1986, and reported in 1993 that mutation of the gc chain results in X-linked severe combined immunodeficiency (XSCID, which has a T-B+NK- phenotype) in humans. We reported in 1995 that mutations of the gc-associated kinase, JAK3, result in an autosomal recessive form of SCID indistinguishable from XSCID and in 1998 that T-B+NK+ SCID results from mutations in the IL7R gene. Based on work in our lab and others, gc was previously shown to be shared by the receptors for IL-2, IL-4, IL-7, IL-9, IL-15, and IL-21. We also previously characterized genes that were induced or repressed by IL-2, IL-4, IL-7, and IL-15, including showing the negative regulation of the IL-7 receptor alpha chain, a finding with implications in understanding how IL-2 can promote cell death, and the positive regulation of a dual specificity phosphatase, DUSP5, that negatively regulates IL-2-mediated activation of ERK kinases. T helper cell differentiation is critical for normal immune responses, with Th1 differentiation being important for host defense to viruses and other intracelllular pathogens, Th2 differentiation being vital in allergic disorders and related to helminths, and Th17 differentiation being vital in a range of inflammatory disorders, including psoriasis and inflammatory bowel disease. We previously showed that IL-2 is important for Th2 differentiation and reported that IL-2 regulates expression of the IL-4 receptor in a STAT5-dependent manner and critically controls priming of cells for Th2 differentiation. Moreover, using genome-wide Illumina-based ChIP-Seq (chromatin immunoprecipitation coupled to DNA sequencing) analysis, we had discovered broad regulation of Th2 differentiation via STAT5A and STAT5B, substantially extending earlier studies focused on STAT5A. Moreover, we had discovered that IL-2-mediated IL-4Ra induction was critical in priming cells for Th2 differentiation. In the prior year, we substantially extended these findings by showing that IL-2 via STAT5 induces expression of IL-12Rb1 and IL-12Rb2 and that the induction of IL-12Rb2 is critical for Th1 differentiation and we defined the mechanism of regulation of IL-12Rb2. Additionally, we showed that IL-2 via STAT5 also regulates the T box protein, T-bet. Interestingly, in contrast to the induction of IL-12R proteins, IL-2 inhibits expression of IL-6Ra and gp130, helping to explain the inhibition of Th17 differentiation. Consistent with the ability of Tbx21 to inhibit Th17 differentiation, expression of Tbx21 in Th17 cells resulted in increased IFNg but decreased expression of IL-17A. These results indicated a very broad effect of IL-2 via STAT5 on T helper cell differentiation. In the current review year, we have continued to study the role of IL-2 in Th differentiation, extending findings to Th9 differentiation. We also have continued our studies of the role of STAT5 tetramerization in vivo, with our initial report having been in the prior year. During the past year, we collaborated with Dr. K. Christopher Garcia at Stanford, studying the actions of wild type IL-2 versus novel IL-2 variants, a project with potential clinical ramifications. These studies in part use the pmel-1 T cell receptor transgenic model of adoptive immunotherapy for cancer in collaboration with Dr. Nicholas Restifo, NCI. We also collaborated with Dr. Garcia on a project in which the three dimensional structure of IL-2 complexed to its receptor was compared to that of IL-15 bound to its receptor. These studies, reported in the past year, have provided key mechanistic and structural insights into the functional differences between IL-2 and IL-15, which are highly related and share IL-2Rbeta and gc as receptor components but nevertheless possess distinctive biological functions. Although IL-2 primarily signals via cis-signaling and IL-15 via trans-signaling, these cytokines have essentially identical activation of STAT, PI3K/Akt, and Ras/MAPK signaling pathways. Moreover, gene expression profiles are very similar, although not identical. Thus, these cytokines have almost indistinguishable signaling properties despite different biological responses. This study has substantially elucidated structural and mechanistic aspects of IL-2 and IL-15 signaling. We previously reported the generation of IL-2/IL-21 dual reporter BAC transgenic mice. These mice allow one to observed through surrogate fluorescent markers the relative induction of these cytokines in different cells. Using these mice, we previously demonstrated a key role for IL-21 in a model mouse system for experimental autoimmune uveitis, and we had demonstrated the existence of IL-2, IL-21, and IL-2/IL-21 double expressing cells in the retina, consistent with a role for this cytokine in the disease process. We are continuing to develop studies with these valuable animals, with the intent of learning more about the roles of IL-2 and IL-21 in vivo. We also have the long-term goal of identifying new causes of human immunodeficiency and other hematological diseases. Overall, these studies help to improve our understanding of signaling by gc family cytokines. These findings clarify basic molecular mechanisms that are relevant to normal and pathological immune cell function, including allergy, autoimmunity, and cancer.

IL-2 受体和相关细胞因子受体系统正在研究中，以阐明正常、肿瘤和免疫缺陷状态下的 T 细胞免疫反应。 T 细胞被抗原激活后，T 细胞免疫反应的强度和持续时间取决于产生的 IL-2 量、表达的受体水平以及每个事件的时间进程。 IL-2受体包含3条链：IL-2Ra、IL-2Rb和gc。 Leonard 博士于 1984 年克隆了 IL-2Ra，我们于 1986 年发现了 IL-2Rb，并于 1993 年报道了 gc 链突变导致 X 连锁严重联合免疫缺陷（XSCID，具有 T-B+NK- 表型）在人类中。我们于 1995 年报道，gc 相关激酶 JAK3 的突变导致与 XSCID 无法区分的常染色体隐性 SCID，并于 1998 年报道 T-B+NK+ SCID 由 IL7R 基因突变引起。根据我们实验室和其他实验室的工作，gc 先前被证明由 IL-2、IL-4、IL-7、IL-9、IL-15 和 IL-21 受体共享。我们之前还鉴定了 IL-2、IL-4、IL-7 和 IL-15 诱导或抑制的基因，包括显示 IL-7 受体 α 链的负调节，这一发现对于理解 IL-2 -2 可以促进细胞死亡，并正向调节双特异性磷酸酶 DUSP5，从而负向调节 IL-2 介导的 ERK 激酶激活。 T辅助细胞分化对于正常免疫反应至关重要，Th1分化对于宿主防御病毒和其他细胞内病原体很重要，Th2分化在过敏性疾病和与蠕虫相关的疾病中至关重要，而Th17分化在一系列炎症性疾病中至关重要，包括牛皮癣和炎症性肠病。我们之前表明IL-2对于Th2分化很重要，并报道IL-2以STAT5依赖性方式调节IL-4受体的表达，并关键控制细胞的Th2分化启动。此外，使用基于 Illumina 的全基因组 ChIP-Seq（染色质免疫沉淀与 DNA 测序相结合）分析，我们发现 STAT5A 和 STAT5B 对 Th2 分化有广泛的调节，从而大大扩展了早期针对 STAT5A 的研究。此外，我们发现 IL-2 介导的 IL-4Ra 诱导对于启动细胞进行 Th2 分化至关重要。去年，我们进一步扩展了这些发现，表明 IL-2 通过 STAT5 诱导 IL-12Rb1 和 IL-12Rb2 的表达，并且 IL-12Rb2 的诱导对于 Th1 分化至关重要，并且我们定义了 IL-2 的调节机制-12Rb2。此外，我们还发现 IL-2 通过 STAT5 还可以调节 T 盒蛋白 T-bet。有趣的是，与IL-12R蛋白的诱导相反，IL-2抑制IL-6Ra和gp130的表达，有助于解释Th17分化的抑制。与 Tbx21 抑制 Th17 分化的能力一致，Th17 细胞中 Tbx21 的表达导致 IFNg 增加，但 IL-17A 表达减少。这些结果表明 IL-2 通过 STAT5 对 T 辅助细胞分化产生非常广泛的影响。在当前的回顾年中，我们继续研究 IL-2 在 Th 分化中的作用，并将研究结果扩展到 Th9 分化。我们还继续研究 STAT5 四聚化在体内的作用，我们的初步报告是在去年。去年，我们与斯坦福大学的 K. Christopher Garcia 博士合作，研究野生型 IL-2 与新型 IL-2 变体的作用，这是一个具有潜在临床影响的项目。这些研究部分使用了与 NCI 的 Nicholas Restifo 博士合作的癌症过继免疫疗法的 pmel-1 T 细胞受体转基因模型。我们还与 Garcia 博士合作开展了一个项目，其中将与其受体复合的 IL-2 的三维结构与与其受体结合的 IL-15 的三维结构进行比较。去年报道的这些研究为 IL-2 和 IL-15 之间的功能差异提供了关键的机制和结构见解，它们高度相关，并共享 IL-2Rbeta 和 gc 作为受体成分，但仍然具有独特的生物学功能。尽管 IL-2 主要通过顺式信号传导而 IL-15 主要通过反式信号传导发出信号，但这些细胞因子对 STAT、PI3K/Akt 和 Ras/MAPK 信号传导途径的激活作用基本相同。此外，基因表达谱虽然不相同，但非常相似。因此，尽管生物反应不同，但这些细胞因子具有几乎无法区分的信号传导特性。这项研究充分阐明了 IL-2 和 IL-15 信号传导的结构和机制。我们之前报道过 IL-2/IL-21 双报告基因 BAC 转基因小鼠的产生。这些小鼠允许通过替代荧光标记观察这些细胞因子在不同细胞中的相对诱导。我们之前使用这些小鼠证明了 IL-21 在实验性自身免疫性葡萄膜炎模型小鼠系统中的关键作用，并且我们还证明了 IL-2、IL-21 和 IL-2/IL-21 双表达细胞的存在在视网膜中，与该细胞因子在疾病过程中的作用一致。我们正在继续对这些有价值的动物进行研究，目的是更多地了解 IL-2 和 IL-21 在体内的作用。我们的长期目标是确定人类免疫缺陷和其他血液疾病的新病因。总的来说，这些研究有助于提高我们对 gc 家族细胞因子信号传导的理解。这些发现阐明了与正常和病理免疫细胞功能相关的基本分子机制，包括过敏、自身免疫和癌症。