NKT cells activated by apoptotie cells through TIM-1 regulate asthma

凋亡细胞通过 TIM-1 激活 NKT 细胞调节哮喘

• 批准号: 8507122
• 负责人: DALE T UMETSU
• 金额: $ 31.69万
• 依托单位: BOSTON CHILDREN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-09-30 至 2014-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8507122
• 关键词: Affect; Apoptosis; Apoptotic; Asthma; Binding; Cell Line; Cell physiology; Cells; Development; Disease; Environment; Epithelial Cells; General Population; Goals; Grant; Hepatic; Hepatitis; Hepatitis A Virus; Hepatocyte; Human; Hypersensitivity; Inflammation; Lead; Liver; Lung; Macaca mulatta; Mediating; Modeling; Molecular; Monkeys; Mus; Natural Immunity; Oxidative Stress; Pathway interactions; Pattern; Pattern recognition receptor; Phosphatidylserines; Public Health; Reagent; Signal Transduction; Susceptibility Gene; Virus Diseases; adaptive immunity; airway hyperresponsiveness; airway inflammation; atopy; cytotoxic; in vivo; in vivo Model; insight; killer T cell; novel; phosphatidylserine receptor; receptor

The long-term goals of Project 1 are to determine how TIM-1, which binds phsophatidylserine (PtdSer) on apoptotic cells, regulates the development of asthma. In the previous grant period, we showed that TIMl is an important atopy susceptibility gene interacting with the envirormient, and that TIM-1 is an important receptor for PtdSer. In the next grant period, we will extend these observations, and hypothesize that asthma is regulated by Natural Killer T (NKT) cells expressing TIM-1, and activated by apoptotic bronchial epithelial cells expressing PtdSer. We will show that NKT cells activated by apoptotic cells in the airways can amplify innate and adaptive immunity, and cause airway hyperreactivity (AHR), a cardinal feature of asthma. In Specific Aim 1, we propose to clarify the mechanisms by which TIM-1 costimulates the activation of NKT cells. We will demonstrate that apoptotic cells bind to and activate NKT cells, in a TIM-1 and PtdSer specific manner. Moreover, we will show that such a pathway can occur in vivo, by establishing an in vivo model in which liver cells, made apoptotic by treatment with anti-Fas mAb, activate NKT cells, which are present in large numbers in the liver. In Specific Aim 2, we hypothesize that the hepatitis A virus (HAV), by binding to TIM-1 on human NKT cells, can activate the NKT cells. We will show that that HAV-activated NKT cells are cytotoxic for hepatocytes, and suggest that these HAV-activated NKT cells later protect against the development of asthma. We will also show that TIMl, a susceptibility gene for asthma, is also a susceptibility gene for severe HAV infection and hepatitis. Moreover, by examining a monkey model of HAV infection, we will demonstrate that HAV infection is indeed associated with the activation and expansion of hepatic NKT cells. In Specific Aim 3, we will examine how NKT cells, responding through TIM-1 to apoptotic airway epithelial cells in the lung, mediate AHR. We hypothesize that oxidative stress in the airways causes airway epithelial cell apoptosis, which can then activate NKT cells, resulting in the development of AHR. Our studies will be facilitated by unique reagents, including primary NKT cell lines, activating and blocking anti-TIM-1 mAb, TIM-1 Tg mice, TIM -/- mice, TIM-3-/- mice, Nrf2 -/- mice and Rhesus monkeys. These studies will provide significant insight into an important human atopy susceptibility gene (TIMl). Moreover, we will demonstrate that TIM-1 on NKT cells functions as a pattern recognition receptor that senses PtdSer as a DAMP (damage associated molecular pattern), and that apoptotic bronchial epithelial cells provide a "danger" signal that activates NKT cells, profoundly affecting airway inflammation and asthma.

项目1的长期目标是确定TIM-1如何与凋亡细胞上的磷脂酰丝氨酸（PtdSer）结合，调节哮喘的发展。在前期研究中，我们发现TIM-1是一个与环境相互作用的重要特应性易感基因，TIM-1是PtdSer的重要受体。在下一个资助期，我们将扩展这些观察，并假设哮喘是由表达TIM-1的自然杀伤T（NKT）细胞调节的，并由表达PtdSer的凋亡支气管上皮细胞激活。我们将证明气道中凋亡细胞激活的NKT细胞可以放大先天性和适应性免疫，并导致气道高反应性（AHR），这是哮喘的一个主要特征。 在具体目标1中，我们建议阐明TIM-1共刺激激活 NKT细胞。我们将证明凋亡细胞以TIM-1和PtdSer特异性方式结合并激活NKT细胞。此外，我们将表明，这样的途径可以在体内发生，通过建立一个体内模型，其中肝细胞，通过与抗Fas单克隆抗体治疗凋亡，激活NKT细胞，这是在肝脏中大量存在。 在特异性目的2中，我们假设甲型肝炎病毒（HAV）通过与人类细胞表面的TIM-1结合， NKT细胞，可以激活NKT细胞。我们将证明HAV激活的NKT细胞具有细胞毒性， 肝细胞，并表明这些HAV激活的NKT细胞后来保护免受哮喘的发展。 我们还将表明，TIM 1，哮喘的易感基因，也是严重的HAV感染和肝炎的易感基因。此外，通过检查HAV感染的猴模型，我们将证明HAV感染确实与肝NKT细胞的活化和扩增相关。在具体目标3中，我们将研究NKT细胞如何通过TIM-1对肺中凋亡的气道上皮细胞作出反应，介导AHR。我们假设气道中的氧化应激导致气道上皮细胞凋亡，然后激活NKT细胞，导致AHR的发展。 我们的研究将通过独特的试剂来促进，包括原代NKT细胞系，激活和 阻断抗TIM-1 mAb、TIM-1 Tg小鼠、TIM -/-小鼠、TIM-3-/-小鼠、Nrf 2-/-小鼠和恒河猴。 这些研究将为重要的人类特应性易感基因（TIM 1）提供重要的见解。 此外，我们将证明NKT细胞上的TIM-1作为模式识别受体，将PtdSer感知为DAMP（损伤相关分子模式），并且凋亡的支气管上皮细胞提供激活NKT细胞的“危险”信号，深刻影响气道炎症和哮喘。