INTERLEUKIN-11 MEDIATED SIGNAL TRANSDUCTION

INTERLEUKIN-11 介导的信号转导

• 批准号: 3367984
• 负责人: YU-CHUNG YANG
• 金额: $ 15.13万
• 依托单位: INDIANA UNIV-PURDUE UNIV AT INDIANAPOLIS
• 依托单位国家: 美国
• 财政年份: 1992
• 资助国家: 美国
• 起止时间: 1992-08-01 至 1997-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3367984
• 关键词: G protein; adenylate cyclase; arachidonate; autoradiography; biological signal transduction; cell cycle; clone cells; complementary DNA; cyclic AMP; cytokine; cytokine receptors; diacylglycerols; genetic library; genetic mapping; hematopoiesis; hematopoietic growth factor; inositol phosphates; interleukin 3; interleukin 6; molecular cloning; northern blottings; nuclear runoff assay; pertussis toxin; phorbols; phosphatase inhibitor; phosphatidate; phosphorylation; protein tyrosine kinase; receptor binding; regulatory gene; second messengers; transfection /expression vector; western blottings

The cDNA encoding IL-11 was isolated from a primate bone marrow stromal cell line through mammalian cell expression cloning method based on its ability to stimulate the proliferation of an IL-6-dependent mouse plasmacytoma cell line, T1165. Like most of the cytokines, IL-11 exerts various biological activities through the interactions-between the ligand and the receptors present on the target cells. IL-11 has been shown to be able to synergize with IL-3 in supporting human and murine megakaryocyte colony formation, stimulate the T-cell dependent development of immunoglobulin-producing B cells and act synergistically with IL-3 to shorten the Go period of early hematopoietic progenitors. Recent studies have shown that IL-11 can also induce the synthesis of acute-phase proteins and inhibits the process of adipocyte differentiation. The fact that an adipocyte cell line, 3T3-Ll, can respond to a stromal fibroblast-derived growth factor like IL-11, may .provide a useful system to study the cell-cell interactions in the hematopoietic microenvironment. The long term objective of this proposal is to understand the role of IL-11 in regulating hematopoiesis at the molecular and cellular level. Since most of the biological activities of IL-11 overlaps with those of IL-6, we hypothesize that these two cytokines may utilize similar signal transduction pathways or may even share common receptors. The proposed work aims to (1) molecularly clone the IL-11 receptor: this will be achieved by (i) cloning the receptor based on the functional interaction between the i ligand and the receptor by screening a 3T3-Ll cDNA expression library based on the ability of the IL-11 receptor cDNA expressing IL-3-dependent FDCP-1 cells to grow in the presence of IL-11 and in the absence of IL-3; (ii) cloning the receptor based on the ligand binding by screening a 3T3-Ll cDNA expression library using microscopic emulsion autoradiography with radioiodinated IL-11 and (2) study IL-11-mediated signal transduction pathways in comparison with those of IL-6: this will be accomplished by (i) analyzing the significance of protein tyrosine phosphorylation, G protein, protein kinase C and adenyl cyclase in IL11-mediated sinal transduction pathways; (ii) identifying phospholipid-derived second messengers involved in the IL-11-mediated signaling; (iii) examining the primary response genes activated by IL-11-mediated signal transduction. The accomplishment of this proposal will not only help us better understand the biological relevance of IL-11 and cell-cell interactions in the normal hematopoietic microenvironment but also uncover some of the mechanisms involved in the IL-11-mediated signal transduction.

从灵长类的骨髓基质中分离出编码IL-11的cDNA 通过哺乳动物细胞表达克隆方法基于其细胞系 刺激IL-6依赖性小鼠的增殖的能力 浆细胞瘤细胞系，T1165。 像大多数细胞因子一样，IL-11施加 通过配体之间的相互作用的各种生物学活动 以及靶细胞上存在的受体。 IL-11已显示为 能够与IL-3协同支持人和鼠 巨核细胞菌落形成，刺激T细胞依赖性 开发产生免疫球蛋白的B细胞并协同作用 IL-3可以缩短早期造血祖细胞的期限。 最近的研究表明，IL-11也可以诱导合成 急性期蛋白质并抑制脂肪细胞的过程 分化。 脂肪细胞细胞系3T3-​​ll可以 响应基质成纤维细胞衍生的生长因子，例如IL-11，可能 。 造血微环境。 该提议的长期目标 是要了解IL-11在调节造血中的作用 分子和细胞水平。 由于大多数生物学活动 IL-11与IL-6重叠，我们假设这两个 细胞因子可能使用类似的信号转导途径，甚至可能 共享共同的受体。 提出的工作旨在（1）分子克隆 IL-11受体：这将通过（i）克隆受体来实现 基于I配体与受体之间的功能相互作用 通过筛选3T3-LL cDNA表达式库 IL-11受体cDNA表达IL-3依赖性FDCP-1细胞生长在 IL-11的存在，在不存在IL-3的情况下； （ii）克隆受体 基于配体结合，通过筛选3T3-LL cDNA表达式文库 使用微观乳液放射自显影和放射性固定的IL-11和 （2）研究IL-11介导的信号转导途径与 IL-6的那些：这将通过（i）分析来完成 蛋白酪氨酸磷酸化，G蛋白，蛋白质的重要​​性 IL11介导的辛纳尔转导途径中的激酶C和腺基环化酶； （ii）识别参与磷脂的第二个使者 IL-11介导的信号传导； （iii）检查主要响应基因 被IL-11介导的信号转导激活。 成就 该建议不仅将帮助我们更好地了解生物学 IL-11和细胞 - 细胞相互作用在正常造血的相关性 微环境，但也发现了涉及的一些机制 IL-11介导的信号转导。