Modeling influenza virus replication in primary human lung cells

模拟流感病毒在原代人肺细胞中的复制

• 批准号: 7241240
• 负责人: FREDERICK T KOSTER
• 金额: $ 30.59万
• 依托单位: LOVELACE BIOMEDICAL & ENVIRONMENTAL RESEARCH INSTITUTE
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-04-15 至 2009-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7241240
• 关键词: Accounting; Adherent Culture; Agonist; Air; Alveolar; Apical; Attention; Avian Influenza; Biological; Birds; Cataloging; Catalogs; Cells; Collectins; Computer Simulation; Computer software; Data; Diffusion; Elements; Environment; Epithelial; Epithelial Cells; Event; Goals; Growth; Human; Immune response; Immunohistochemistry; In Vitro; Indium; Individual; Infection; Influenza; Influenza A Virus, H5N1 Subtype; Interferon Activation; Kinetics; Label; Lateral; Liquid substance; Location; Lung; Measures; Mediating; Metric; Modeling; Molecular Biology; Mucins; Neuraminidase; Numbers; Oseltamivir; Outcome; Pathogenesis; Pattern; Peptides; Persons; Phenotype; Plaque Assay; Production; Productivity; Protein Array; Public Health; Pulmonary Surfactant-Associated Protein A; Rate; Relative (related person); Relative Risks; Research; Respiratory System; Structure; Surface; Testing; Time; Tissues; Tracheobronchial; Viral; Virion; Virulence; Virus; Virus Diseases; Virus Receptors; Virus Replication; Work; absorption; apical membrane; base; design; extracellular; flu transmission; human data; human tissue; immortalized cell; in vitro Model; in vivo Model; influenzavirus; inhibitor/antagonist; pandemic disease; pathogen; receptor binding; reproductive; research study; simulation; spatial relationship; surfactant; transmission process; two-dimensional; vaccine development

DESCRIPTION (provided by applicant): Description Concern about a human pandemic of highly pathogenic avian (H5N1) influenza has focused attention on several related problems: understanding the pathogenesis and transmission of influenza A infections, pre-pandemic recognition of potential pandemic strains, and pre-pandemic vaccine development. Although molecular biology has provided a detailed understanding of the replication cycle in immortalized cells, influenza replication at the intact tissue level among phenotypically diverse epithelial cells of the human respiratory tract remains poorly understood. We are missing the quantitative kinetics accounting in the human airway and how one strain, but not a closely related strain, can initiate person-to-person transmission. We propose to model early influenza virion productivity in human airway epithelial cells. The in vitro model uses primary human differentiated tracheobronchial and bronchiolo-alveolar epithelial cells grown in air-liquid interface (ALI) culture to document the kinetics of virion productivity of human pathogens, both H3N2 strains and H5N1 avian influenza strains. The agent-based computer model is a two-dimensional cellular automata style simulation, designed and implemented by one of us (CB). Kinetic data collected from the in vitro model will be used validate the details and parameter values of the computer model, and then the computer model will be used to study unmeasurable variables and outcomes in the in vitro model. The goal of the project is to use experimentally derived kinetic data of influenza virus replication in primary human lung epithelial cells to calculate metrics of viral growth, particularly the Ro (reproductive rate, average number of secondary infected cells produced by one primary cell), in order to obtain a human tissue-relevant indicator of virulence and transmissibility, capable of distinguishing strains without expensive in vivo models. Using data from the in vitro model, we will determine the following parameters in the computer model: unit rate of viral production per cell, virion diffusion rate, and the unit rate of virion absorption by epithelial cells. The kinetics of strains of H3N2 human pathogenic influenza A will be compared to those of selected human pathogenic H5N1 avian influenza strains, both experimentally and in the computer model. We will manipulate the environment of the ALI culture experimentally to augment or diminish components of the innate host response including mucins and surfactant collectins. Analogous manipulations will be implemented in the computer model and results compared between the two models for all the different strains. We will then use the agent-based model to systematically study the relative contribution of each innate mucosal defense element, both in terms of individual and collaborative impacts on the R0 for each influenza strain. This project develops a test designed to distinguish threatening bird flu strains from non-threatening strains. The test measures influenza virus growth in cultured mature human lung airway cells and uses mathematical analysis to calculate the spread of the virus through the culture. Bird flu is considered by many to be potentially the next greatest threat to public health in the world.

描述（由申请人提供）：描述对高度致病性禽（H5N1）流感的人类大流行的关注，将注意力集中在几个相关问题上：了​​解流感和传播感染的发病和传播，对潜在大流血菌株的大流行识别，以及perdecial病前疫苗的发育。尽管分子生物学对永生细胞中的复制周期提供了详细的了解，但人类呼吸道的表型多样性上皮细胞之间完整的组织水平的流感复制仍然很众所周知。我们缺少人类气道中的定量动力学会计，以及一种菌株（而不是密切相关的压力）如何引发人与人的传播。我们建议在人气道上皮细胞中建模早期流感病毒体生产力。体外模型使用在空气界面（ALI）培养物中生长的原发性人体分化的气管和支气管 - 肺泡上皮细胞，以记录人类病原体的病毒体生产力，H3N2菌株和H5N1 AVIAN流感菌株的病毒性生产力。基于代理的计算机模型是由我们一个人（CB）设计和实施的二维蜂窝自动机仿真模拟。将使用从体外模型中收集的动力学数据验证计算机模型的详细信息和参数值，然后计算机模型将用于研究体外模型中不可衡量的变量和结果。该项目的目的是使用人类原代人肺上皮细胞中流感病毒复制的实验得出的动力学数据来计算病毒生长的指标，尤其是RO（生殖速率，一个原代细胞产生的二次感染细胞的平均数量），以使人体组织的模型具有较高的能力，以使能力稳定的能力稳定性，以使能力稳定。使用来自体外模型的数据，我们将确定计算机模型中的以下参数：每个细胞的病毒生产速率，病毒体扩散速率和上皮细胞的病毒体吸收率。 H3N2人类致病性流感菌株的动力学将与精选的人类致病性H5N1鸟类流感菌株的动力学相提并论，无论是在实验和计算机模型中。我们将通过实验来操纵ALI培养的环境，以增强或减少先天宿主反应的组成部分，包括粘蛋白和表面活性剂集群。类似的操作将在计算机模型中实现，并在所有不同菌株的两个模型之间进行比较。然后，我们将使用基于代理的模型来系统地研究每个先天粘膜防御元件的相对贡献，无论是个人和协作对每种流感菌株R0的影响而言。该项目开发了一项旨在区分威胁性鸟类流感菌株和无威胁性菌株的测试。该测试测量培养成熟的人肺气道细胞中流感病毒的生长，并使用数学分析来计算病毒通过培养物的扩散。许多人认为鸟流感可能是对世界上公共卫生的下一个最大威胁。