In Vitro and in Vivo Lung Studies

这个细胞培养和动物核心的设计是为了支持本项目中4个项目中的每个项目的具体目标 修订后的计划项目申请。有两个具体目标。第一个目标是供应肺泡上皮 来自人肺、大鼠肺和小鼠肺的II型细胞，以支持项目1(莫斯托夫)的特定目标， 项目2(Engel)和项目3(Rosen)。我们的研究小组在隔离方面有丰富的经验， 人和啮齿动物肺泡上皮II型细胞的培养和研究 正常和病理状态。这项工作在研究人类免疫缺陷的机制方面尤其富有成效。 铜绿假单胞菌ExoT和ExOS对肺泡上皮细胞的损伤以及ADP核糖如何 铜绿假单胞菌ExoT转移酶结构域与其生物活性的关系 生物体(项目2，恩格尔)。此外，我们最近的工作已经证实，人类肺泡上皮细胞 II型细胞可以在基质细胞中培养，并被诱导形成类似真实细胞的三维结构 肺泡状结构。这一成就为肺的新研究提供了一个重大机遇。 正常和病理条件下的上皮功能(项目1，莫斯托夫)。这个核心的第二个目标， 这代表了一个扩展的方向，就是利用小鼠的病理模型进行体内研究 肺损伤以检验项目3(ROSEN)和项目4(WERB)中的特定假设。这些活体小鼠 实验将使用感染性和非感染性刺激来测试对肺功能和总体的影响 生死存亡。我们的研究小组在活体研究方面有丰富的经验，有相当多的 在测试特定假设时生成的初步数据，即项目3的一部分(罗森)和 项目4(WERB)。

This Cell Culture and Animal Core is designed to support specific aims in each of the 4 projects in this revised Program Project Application. There are two specific aims. The first aim is to supply alveolar epithelial type II cells from human lungs, rat lungs, and mouse lungs to support the specific aims of Project 1 (Mostov), Project 2 (Engel), and Project 3 (Rosen). Our research group has had extensive experience in isolating, culturing, and studying pulmonary alveolar epithelial type II cells from human and rodent lungs under both normal and pathological conditions. This work has been especially productive in studying mechanisms of alveolar epithelial injury from P.aeruginosa ExoT and ExoS as well as establishing how the ADP ribosyl transferase domain of P.aeruginosa ExoT contributes to the biologic activities of the Pseudomonas organisms (Project 2, Engel). In addition, our recent work has established that human alveolar epithelial type II cells can be cultured in matrigel and induced to form 3-dimentional structures that resemble authentic alveolar-like structures. This accomplishment provides a major opportunity for novel studies of lung epithelial function under normal and pathologic conditions (Project 1, Mostov). The second aim of this core, which represents an expanded direction, is to carry out in vivo mouse studies using pathologic models of lung injury to test specific hypotheses in Project 3 (Rosen) and Project 4 (Werb). These in vivo mouse experiments will use both infectious and non-infectious stimuli to test the effects on lung function and overall survival. Our research group has extensive experience with in vivo studies and there is considerable preliminary data that has been generated in testing specific hypotheses that a part of Project 3 (Rosen) and Project 4 (Werb).