Cytolytic Attack Against Lung Parenchyma in Emphysema

描述（由申请人提供）： 许多无症状的HIV感染者发展为肺泡炎， 肺气肿，但发病机制仍然不明。 我们发现细胞毒素 淋巴细胞（CTL）在发展成淋巴瘤的受试者的气道中升高， 肺气肿，这些患者有大量的形态学 实质组织缺损的证据。 这证实了其他 研究表明，CTL升高与早期下降同时发生， 扩散能力和较低气道上皮通透性的增加。 一 关键的观察结果是，从HIV感染者的肺中分离的CTL 可以完成MHC 1类限制性杀死HIV感染细胞。 的 这次调查的重点是扩大一系列突破性的 申请人的发现揭示了以下内容，1）141 V感染 人肺上皮细胞在体内和体外; 2）肺 上皮允许HIV复制;和3）HIV 肺上皮细胞感染诱导CTL介导的MHC凋亡 限制的方式。 具体目标1将检验以下假设： 原代人肺上皮细胞被HIV-1从顶端感染， 基底外侧表面并允许病毒在细胞（NF-0）后复制 activation. 我们预测，这将导致释放有能力的自由 病毒 具体目标2将检验以下假设： 人肺上皮细胞触发CTL介导的攻击， 上皮细胞凋亡 在这一追求中，我们将通过以下方式确定机制： 发生哪种上皮细胞凋亡，然后确定 上皮细胞有丝分裂原，角质细胞生长因子，可以防止肺 上皮细胞死亡。 在修订后的应用程序中，我们将开发一个生理相关的模型 用于艾滋病研究。 特别地，原代人肺上皮细胞将 在胶原包被的跨孔插入物上生长， 在空气-液体界面处具有紧密连接的分化的单层。 该模型将被用来确定优先艾滋病毒运输，我们相信， 由于上皮细胞可作为HIV进入的门户， 进入肺（基底外侧到顶端），并将病毒重新引入肺内。 循环（顶侧至基底侧）。 后续研究将利用 相同的模型，以确定自体，HIV特异性，CTL如何处理HIV+ 肺上皮细胞 申请人的长期目标是建立一个 在翻译研究的职业生涯，连接他目前的专业知识， 临床医学与基础科学 候选人已经做出了实质性的 致力于从事探索肺实质反应的职业 并将发现转化为对患者有用的解决方案 肺部疾病 申请人已建立了一个优秀的团队， 顾问，以协助在这奋进，并已获得适当的 机构支持，以完成培训。

DESCRIPTION (provided by applicant): Many asymptomatic HIV-infected individuals develop alveolitis followed by emphysema but the pathogenesis remains unknown. We have found that cytotoxic lymphocytes (CTLs) are elevated in the airway of subjects that develop emphysema and that these patients have substantial morphologic evidence of parenchymal tissue loss. This corroborates other studies demonstrating that CTL elevation coincides with an early decline in diffusion capacity and an increase in lower airway epithelial permeability. A key observation is that CTLs isolated from the lung of HIV-infected subjects can accomplish MHC class 1 restricted killing of HIV infected cells. The focus of this investigation is to expand on a series of ground-breaking discoveries made by the applicant that reveal the following, 1) 141V infects human lung epithelial cells in vivo and in vitro; 2) the lung epithelium is permissive to HIV replication; and 3) that HIV infection of lung epithelial cells induces CTL-mediated apoptosis in an MHC restricted manner. Specific Aim 1 will test the hypothesis that primary human lung epithelial cells are infected by HIV-1 from the apical and basolateral surface and permit viral replication following cell (NF-0) activation. We predict that this will result in the release of competent free virus. Specific Aim 2 will test the hypothesis that HIV infection of primary human lung epithelial cells triggers a CTL-mediated assault resulting in epithelial cell apoptosis. In this pursuit we will identify the mechanism by which epithelial cell apoptosis occurs and then determine if a potent epithelial cell mitogen, keratinocyte growth factor, can prevent lung epithelial cell death. In the revised application we will develop a physiologically relevant model for studies with HIV. In particular, primary human lung epithelial cells will be grown on collagen coated trans-well inserts to establish fully differentiated monolayers with tight junctions at an air-liquid interface. The model will be used to identify preferential HIV transport which we believe is highly relevant since the epithelium may serve as a portal for HIV entry into the lung (basolateral to apical) as well as reintroduce virus back into the circulation (apical to basolateral). Subsequent studies will utilize the same model to identify how autologous, HIV-specific, CTLs dispose of the HIV+ lung epithelial cell. The long-term goal of the applicant is to establish a career in translational research that connects his current expertise in clinical medicine with basic science. The candidate has made a substantial commitment to pursue a career that will explore the lung parenchymal response to inflammation and translate discoveries into useful solutions for patients with lung disease. The applicant has established an excellent group of advisors to assist in this endeavor and has secured the appropriate institutional support to complete the training.