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

• 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
• 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

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.

正在研究IL-2受体和相关的细胞因子受体系统，以阐明正常，肿瘤和免疫缺陷态的T细胞免疫反应。通过抗原激活T细胞后，T细胞免疫反应的大小和持续时间由产生的IL-2量，表达的受体水平以及每个事件的时间过程确定。 IL-2受体包含三个链IL-2RA，IL-2RB和GC。伦纳德（Leonard）博士于1984年克隆了IL-2RA，我们在1986年发现了IL-2RB，并在1993年报道说，GC链的突变导致X链的严重合并免疫缺陷（XSCID（XSCID），在人类中具有t-b+nk-nk-nk-emotype）。我们在1995年报道说，与GC相关激酶JAK3的突变导致SCID的常染色体隐性形式与XSCID无法区分，并在1998年无法区分t-b+ NK+ SCID是由IL7R基因中的突变引起的。根据我们实验室和其他实验室的工作，GC先前被IL-2，IL-4，IL-7，IL-9，IL-9，IL-15和IL-21的受体共享。 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辅助细胞分化对于正常的免疫反应至关重要，Th1的分化对于宿主防御对病毒和其他细胞内病原体至关重要，Th2分化在过敏性疾病中至关重要，并且与舵机有关，TH17分化对于包括牛皮癣和炎症性弓箭疾病在内的炎症性疾病中至关重要。我们先前表明，IL-2对于TH2分化很重要，并报告IL-2以STAT5依赖性方式调节IL-4受体的表达，并严格控制细胞的TH2分化启动。此外，使用基于全基因组光明的芯片seq（染色质免疫沉淀与DNA测序结合）分析，我们发现了通过STAT5A和STAT5B对TH2分化的广泛调控，从而大大扩展了针对STAT5A的早期研究。此外，我们发现IL-2介导的IL-4RA诱导对于Th2分化的启动细胞至关重要。在上一年中，我们通过表明IL-2通过STAT5诱导IL-12RB1和IL-12RB2的表达来大大扩展了这些发现，并且IL-12RB2的诱导对于TH1的分化至关重要，我们定义了IL-12RB2调节的机制。此外，我们表明IL-2通过STAT5还调节T盒蛋白T-bet。有趣的是，与诱导IL-12R蛋白的诱导相反，IL-2抑制了IL-6RA和GP130的表达，有助于解释Th17分化的抑制作用。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的表达降低。这些结果表明IL-2通过STAT5对T辅助细胞分化的影响非常广。 在当前的审查年中，我们继续研究IL-2在TH分化中的作用，将发现扩展到TH9分化。我们还继续研究体内STAT5四聚体的作用，我们的初步报告已在上一年。 在过去的一年中，我们与斯坦福大学的K. Christopher Garcia博士合作研究了IL-2型与新型IL-2变体的行为，这是一个潜在的临床后果的项目。这些研究部分使用了与NCI的Nicholas Restifo博士合作的PMEL-1 T细胞受体转基因模型的癌症。我们还与Garcia博士合作进行了一个项目，在该项目中，将IL-2的三维结构与其受体相比，将其与其受体结合的项目进行了比较。这些研究在过去一年中报道，对IL-2和IL-15之间的功能差异提供了关键的机理和结构见解，这些功能差异高度相关，并将IL-2RBETA和GC作为受体成分共享，但仍然具有独特的生物学功能。尽管IL-2主要通过顺式信号和IL-15信号通过反式信号，但这些细胞因子基本上具有STAT，PI3K/AKT和RAS/MAPK信号通路的STAT基本相同激活。此外，基因表达谱非常相似，尽管并不相同。因此，尽管生物学反应不同，这些细胞因子几乎具有几乎无法区分的信号传导特性。这项研究已实质上阐明了IL-2和IL-15信号的结构和机械方面。 我们先前报道了IL-2/IL-21双重报告基因BAC转基因小鼠的产生。这些小鼠可以通过替代荧光标记观察到这些细胞因子在不同细胞中的相对诱导。使用这些小鼠，我们先前在用于实验性自身免疫性葡萄膜炎的模型小鼠系统中证明了IL-21的关键作用，我们已经证明了IL-2，IL-21和IL-2/IL-2/IL-21双表达细胞在视网膜中的存在，与该细胞因子在疾病过程中的作用一致。我们正在继续与这些有价值的动物进行研究，并有更多关于体内IL-2和IL-21的作用的意图。 我们还具有确定人类免疫缺陷和其他血液学疾病的新原因的长期目标。 总体而言，这些研究有助于提高我们对GC家族细胞因子的信号传导的理解。这些发现阐明了与正常和病理免疫细胞功能有关的基本分子机制，包括过敏，自身免疫性和癌症。