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HIV感染者患肺泡炎，然后 肺气肿，但发病机理仍然未知。 我们发现细胞毒性 在发展的受试者气道中，淋巴细胞（CTL）升高 肺气肿，这些患者具有大量的形态学 实质损失的证据。 这证实了其他 研究表明，CTL升高与早期下降相吻合 扩散能力和下部气道上皮通透性的增加。 一个 关键观察是，从HIV感染受试者的肺中分离出来的CTL 可以完成MHC 1类限制杀死HIV感染细胞的杀害。 这 这项调查的重点是扩展一系列突破性的 申请人发现以下发现，1）141V感染 人体肺上皮细胞在体内和体外； 2）肺 上皮允许艾滋病毒复制； 3）艾滋病毒 肺上皮细胞的感染诱导MHC中CTL介导的凋亡 受限的方式。 具体目标1将检验以下假设 原发性人肺上皮细胞被顶端和 基底外侧表面并允许细胞后病毒复制（NF-0） 激活。 我们预测这将导致释放有能力的自由 病毒。 特定的目标2将检验以下假设：原发性艾滋病毒感染 人肺上皮细胞触发CTL介导的攻击，导致 上皮细胞凋亡。 在这种追求中，我们将通过 发生哪种上皮细胞凋亡，然后确定是否有效 上皮细胞有丝分裂原，角质形成细胞生长因子，可以防止肺 上皮细胞死亡。 在修订的应用中，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，原发性人肺上皮细胞将 在胶原蛋白涂层的反孔插入物上生长以完全建立 在空气界面处具有紧密连接的分化单层。 该模型将用于识别我们认为的优先艾滋病毒运输 高度相关，因为上皮可以作为HIV进入的门户 进入肺（基底外侧），并将病毒重新引入 循环（顶部至基底外侧）。 随后的研究将利用 相同的模型，以确定自体，艾滋病毒特异性，CTL如何处理HIV+ 肺上皮细胞。 申请人的长期目标是建立 转化研究的职业，将他当前的专业知识联系起来 基础科学的临床医学。 候选人做了大量 致力于追求将探索肺实质反应的职业 发炎并将发现转化为患者有用的解决方案 与肺部疾病。 申请人已经建立了一群很棒的 顾问协助这项工作，并确保了适当的 机构支持以完成培训。