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

项目1的长期目标是确定TIM-1如何调节哮喘的发展，TIM-1与凋亡细胞上的磷脂酰丝氨酸(PtdSer)结合。在之前的资助期间，我们证明了TIML是一个与环境相互作用的重要特应性易感基因，而TIM-1是PtdSer的重要受体。在下一个授权期，我们将延长这些观察，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的凋亡的支气管上皮细胞激活。我们将展示由呼吸道中的凋亡细胞激活的NKT细胞可以放大先天免疫和获得性免疫，并导致哮喘的一个基本特征--气道高反应性(AHR)。 在特定的目标1中，我们建议阐明TIM-1共刺激激活的机制。 NKT细胞。我们将证明凋亡细胞以TIM-1和PtdSer特异性的方式与NKT细胞结合并激活。此外，我们将通过建立体内模型来证明这种途径可以在体内发生，在体内模型中，经抗Fas单抗处理而导致凋亡的肝细胞激活NKT细胞，NKT细胞大量存在于肝脏中。 在特定目标2中，我们假设甲型肝炎病毒(HAV)通过与人类TIM-1结合 NKT细胞，可以激活NKT细胞。我们将证明甲型肝炎病毒激活的NKT细胞对 并提示这些甲型肝炎病毒激活的NKT细胞稍后可预防哮喘的发生。 我们还将展示TIML是哮喘的易感基因，也是重型甲型肝炎和肝炎的易感基因。此外，通过研究甲型肝炎病毒感染的猴子模型，我们将证明甲型肝炎病毒感染确实与肝脏NKT细胞的激活和扩张有关。在特定的目标3中，我们将研究NKT细胞如何通过TIM-1对肺内凋亡的呼吸道上皮细胞做出反应，介导AHR。我们推测，呼吸道氧化应激导致呼吸道上皮细胞凋亡，进而激活NKT细胞，导致AHR的发生。 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 封闭抗Tim-1单抗、Tim-1Tg小鼠、Tim-/-小鼠、Tim-3-/-小鼠、Nrf2-/-小鼠和恒河猴。 这些研究将对一个重要的人类特应性易感基因(TIML)提供重要的见解。 此外，我们将证明NKT细胞上的TIM-1作为模式识别受体发挥作用，将PtdSer感知为潮湿(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，从而深刻影响呼吸道炎症和哮喘。

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.