Cytolytic Attack Against Lung Parenchyma in Emphysema

肺气肿中对肺实质的细胞溶解攻击

• 批准号: 6621326
• 负责人: DAREN Lee KNOELL
• 金额: $ 12.69万
• 依托单位: OHIO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-01-01 至 2006-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6621326
• 关键词: HIV infections; MHC class I antigen; apoptosis; bioassay; cytolysis; cytotoxic T lymphocyte; emphysema; enzyme linked immunosorbent assay; fibroblast growth factor; flow cytometry; gel mobility shift assay; host organism interaction; human tissue; immunocytochemistry; lung; mitogens; model; model design /development; nuclear factor kappa beta; pathologic process; postdoctoral investigator; respiratory epithelium; tissue /cell culture; virus infection mechanism; virus replication

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.

描述（由申请人提供）： 许多无症状的艾滋病毒感染者会出现肺泡炎​​，然后 肺气肿但发病机制仍不清楚。 我们发现细胞毒性 受试者气道中的淋巴细胞（CTL）升高 肺气肿并且这些患者有明显的形态学变化 实质组织损失的证据。 这证实了其他 研究表明，CTL 升高与早期下降相一致 扩散能力和下气道上皮通透性增加。 一个 关键观察结果是从 HIV 感染者的肺部分离出 CTL 可以完成MHC 1类限制性杀伤HIV感染细胞。 这 此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) 141V 感染 体内和体外人肺上皮细胞； 2）肺 上皮细胞允许 HIV 复制； 3）艾滋病毒 肺上皮细胞感染诱导 MHC 中 CTL 介导的细胞凋亡 限制方式。 具体目标 1 将检验以下假设： 原代人肺上皮细胞从顶端和顶端被 HIV-1 感染 基底外侧表面并允许病毒在细胞（NF-0）后复制 激活。 我们预测这将导致有能力的自由释放 病毒。 具体目标 2 将检验原发性 HIV 感染的假设 人肺上皮细胞触发 CTL 介导的攻击，导致 上皮细胞凋亡。 在这一追求中，我们将通过以下方式确定该机制： 哪些上皮细胞发生凋亡，然后确定是否存在有效的细胞凋亡 上皮细胞有丝分裂原、角质细胞生长因子，可预防肺 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模型 用于艾滋病毒研究。 特别是，原代人肺上皮细胞将 在胶原蛋白涂层的跨孔插入物上生长，以充分建立 在气液界面处具有紧密连接的差异化单层。 该模型将用于确定我们认为的优先艾滋病毒运输 高度相关，因为上皮可能作为艾滋病毒进入的门户 进入肺部（从基底外侧到心尖）并将病毒重新引入肺部 循环（心尖至基底外侧）。 后续研究将利用 使用同一模型来确定自体 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 转化研究的职业生涯将他目前的专业知识联系起来 临床医学与基础科学。 候选人做出了重大贡献 致力于探索肺实质反应的职业 炎症并将发现转化为对患者有用的解决方案 患有肺部疾病。 申请人建立了一支优秀的团队 顾问协助这项工作，并已获得适当的 机构支持以完成培训。