Cytolytic Attack Against Lung Parenchyma in Emphysema

描述(由申请人提供)： 许多无症状的艾滋病毒感染者发展为肺泡炎，随后 肺气肿，但发病机制尚不清楚。我们发现它具有细胞毒性 淋巴细胞(CTL)在发育受试者的呼吸道中升高。 肺气肿和这些患者有实质性的形态 实质组织丢失的证据。这证实了其他 研究表明，CTL的上升与早期的下降相一致 扩散能力和较低的呼吸道上皮通透性增加。一个 关键的观察是从HIV感染者的肺中分离出CTL 可完成MHC-1类限制性杀伤HIV感染细胞。这个 此次调查的重点是展开一系列突破性的 申请人的发现表明：1)141V感染 人肺上皮细胞在体内和体外；2)肺 上皮允许HIV复制；以及3)HIV 肺上皮细胞感染诱导巨噬细胞集落刺激因子介导的细胞凋亡 受限的方式。具体目标1将检验以下假设 原代人肺上皮细胞感染HIV-1的根尖细胞和 基底面并允许病毒跟随细胞复制(NF-0) 激活。我们预测，这将导致释放合格的免费 病毒。特定目标2将检验假设，即艾滋病病毒感染的初级 人肺上皮细胞触发CTL介导的攻击导致 上皮细胞凋亡。在这一过程中，我们将通过以下方式确定机制 哪个上皮细胞会发生凋亡，然后确定一个有效的 上皮细胞有丝分裂原，角质形成细胞生长因子，可预防肺 上皮细胞死亡。 在修改后的应用程序中，我们将开发一个与生理相关的模型 用于艾滋病毒的研究。特别是，原代人肺上皮细胞将 生长在胶原蛋白涂层的跨孔插件上，以充分建立 在气-液界面具有紧密连接的差异化单分子层。 该模型将被用来识别我们认为优先的艾滋病毒运输 是高度相关的，因为上皮可能是HIV进入的门户 进入肺(基侧至心尖)，以及将病毒重新引入 循环(心尖至基底外侧)。后续研究将利用 识别自体的、HIV特异性的CTL如何处置HIV+的相同模型 肺上皮细胞。申请者的长期目标是建立一个 在翻译研究方面的职业生涯，将他目前在 具有基础科学的临床医学。这位候选人已经做出了很大的贡献 致力于从事研究肺实质反应的职业 炎症，并将发现转化为对患者有用的解决方案 患有肺病。申请者已经建立了一支优秀的 顾问协助这一努力，并已获得适当的 机构支持，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.