Role of SHP-1 in T cell activation and development

SHP-1 在 T 细胞激活和发育中的作用

• 批准号: 6749457
• 负责人: Ulrike Lorenz
• 金额: $ 33.3万
• 依托单位: UNIVERSITY OF VIRGINIA
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-07-01 至 2006-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6749457
• 关键词: T cell receptor; T lymphocyte; biological signal transduction; cell age; cell growth regulation; cell line; cell membrane; cytotoxic T lymphocyte; embryo /fetus tissue /cell culture; enzyme activity; enzyme mechanism; interleukin 2; laboratory mouse; leukocyte activation /transformation; ligands; peptides; phenotype; protein localization; protein tyrosine phosphatase; receptor expression; thymus; tissue /cell culture; transfection; viral rescue

DESCRIPTION (provided by applicant): Tyrosyl phosphorylation is a key regulatory mechanism for normal cell growth, differentiation, and death. The steady state level of tyrosyl phosphorylation on any protein is determined by he opposing actions of protein tyrosine kinases and phosphatases. SHP-1 is a protein tyrosine phosphatase that has been shown to be involved in the negative regulation of signaling events induced by cytokines, growth factors and antigens. Mutations in the SHP-1 gene in mice cause the motheaten (me/me) phenotype. Mice homozygous for the me allele, which results in the absence of any detectable SHP-1 protein, display a panoply of disorders in 11 hematopoietic lineages. These mice provide a valuable tool to combine in vitro biochemical assays with in vivo and ex vivo biological studies. Our long range goal is to understand how SHP-1 influences the growth and differentiation of hematopoietic cells. This application will attempt to elucidate the involvement of SHP-1 in immature and mature cell functions. Recently, we and others have shown a clear role for SHP-1 in T-cell development. However, the underlying molecular mechanism remains unknown. In the first Aim, we propose to address this question using fetal thymic organ cultures (FTOCs). Analyses of me/me:DO11.10 thymocytes indicate at SHP-1 helps to set TCR signaling thresholds in positive and negative selection. Studies of FTOCs from these mice will be applied to follow thymic T-cell development in detail. In particular, we propose to rescue the motheaten phenotype by reconstituting FTOC from me/me:DO11.10 mice with wild type and mutants of SHP-1 via retroviral gene transfer. While SHP-1' negative regulation of TCR-mediated signaling has been shown, its mechanism of action ha yet to be defined. Recently, one type of specialized membrane microdomains (referred to as lipid-rafts) has been recognized for its importance for TCR signaling. The rafts localization of several key players of early T-cell activation has be n shown to be critical for signaling. Our working hypothesis is that SHP-1 localizes to he rafts and that this localization is important for its function. Indeed, we observe that a fraction of SHP-1 localizes to the rafts. In the second Aim, we propose to study the mechanism of SHP-1's localization to the rafts and its functional consequences. In the third Aim, we will test the hypothesis that SHP-1 plays a role in altered peptide ligand signaling. While SHP-1's role in T-cell development has been recognized, its role in mature cells is less understood. We will use several approaches to address this question with a focus on agonist/partial agonist/antagonist signaling. We will attempt to characterize the responses to altered peptide ligands in wild type and SHP-1-deficient CD4+ DO11.10 TCR-expressing lines as well as CD8+ cytotoxic T-cell clones.

描述（申请人提供）：酪酶磷酸化是关键 正常细胞生长，分化和死亡的调节机制。这 任何蛋白质上酪酶磷酸化的稳态水平由 他反对蛋白酪氨酸激酶和磷酸酶的作用。 SHP-1是一个 蛋白质酪氨酸磷酸酶已被证明与阴性有关 调节细胞因子，生长因子和 抗原。小鼠SHP-1基因中的突变导致motheaten（ME/ME） 表型。 Me纯合子的小鼠为ME等位基因，这导致没有 任何可检测的SHP-1蛋白 造血谱系。这些小鼠提供了一个有价值的工具来结合体外 体内和体内生物学研究的生化测定。我们的远距离 目标是了解SHP-1如何影响 造血细胞。此应用程序将尝试阐明参与 SHP-1的未成熟和成熟细胞功能。 最近，我们和其他人在T细胞中表现出SHP-1的明确作用 发展。但是，基本的分子机制仍然未知。在 第一个目的，我们建议使用胎儿胸腺器官解决这个问题 文化（FTOC）。我/我的分析：do11.10胸腺细胞在SHP-1上指示 将TCR信号传导阈值设置为正选择和负选择。研究 这些小鼠的FTOC将用于遵循胸腺T细胞发育 细节。特别是，我们建议通过 从我/我重新建立ftoc：do11.10小鼠，野生型和SHP-1突变体 通过逆转录病毒基因转移。 虽然已经显示了TCR介导的信号传导的SHP-1'负调控，但 动作机理尚未定义。最近，一种专业 膜微区域（称为脂质生效）已被确认 对于TCR信号的重要性。木筏本地化的几个关键参与者 早期的T细胞激活已被证明对于信号传导至关重要。我们的 工作假设是SHP-1本地化在木筏上，这是 本地化对于其功能很重要。确实，我们观察到这是一部分 SHP-1定位于木筏。在第二个目标中，我们建议研究 SHP-1定位到木筏及其功能后果的机制。 在第三个目标中，我们将检验SHP-1在 改变肽配体信号传导。而SHP-1在T细胞开发中的作用 被认可，其在成熟细胞中的作用尚不清楚。我们将使用 几种方法可以解决这个问题，重点是激动剂/部分 激动剂/拮抗剂信号传导。我们将尝试表征对 野生型和SHP-1缺陷CD4+ DO11.10的肽配体改变了肽配体 表达TCR线以及CD8+细胞毒性T细胞克隆。