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）：描述对人类高致病性禽流感（H5N1）大流行的关注已将注意力集中在几个相关问题上：了解甲型流感感染的发病机制和传播，大流行前对潜在大流行毒株的识别，以及大流行前疫苗的开发。尽管分子生物学已经为永生化细胞的复制周期提供了详细的了解，但人类呼吸道表型不同的上皮细胞在完整组织水平上的流感复制仍然知之甚少。我们缺少人体呼吸道的定量动力学计算，以及一种菌株（但不是密切相关的菌株）如何引发人与人之间的传播。我们建议在人气道上皮细胞中建立早期流感病毒粒子生产力模型。体外模型使用在气液界面（ALI）培养中培养的原代人分化气管支气管和细支气管肺泡上皮细胞来记录人类病原体（H3N2株和H5N1禽流感株）病毒粒子产量的动力学。基于智能体的计算机模型是一种二维元胞自动机式仿真，由我们设计并实现。从体外模型中收集的动力学数据将用于验证计算机模型的细节和参数值，然后计算机模型将用于研究体外模型中不可测量的变量和结果。该项目的目标是利用流感病毒在原代人肺上皮细胞中复制的实验导出的动力学数据来计算病毒生长的指标，特别是Ro（繁殖率，一个原代细胞产生的继发感染细胞的平均数量），以便获得与人体组织相关的毒力和传播率指标，能够在没有昂贵的体内模型的情况下区分菌株。利用体外模型的数据，我们将确定计算机模型中的以下参数：每个细胞的单位病毒产生率、病毒粒子扩散率和上皮细胞对病毒粒子的单位吸收率。H3N2人致病性甲型流感毒株的动力学将通过实验和计算机模型与选定的人致病性H5N1禽流感毒株进行比较。我们将通过实验操纵ALI培养的环境来增加或减少先天宿主反应的成分，包括粘蛋白和表面活性剂集合。将在计算机模型中实施类似的操作，并对所有不同菌株的两个模型之间的结果进行比较。然后，我们将使用基于主体的模型系统地研究每种先天性粘膜防御元素的相对贡献，包括个体和协作对每种流感毒株R0的影响。该项目开发了一种测试，旨在区分具有威胁性的禽流感毒株和不具有威胁性的毒株。该测试测量流感病毒在培养的成熟人肺气道细胞中的生长情况，并使用数学分析来计算病毒在培养物中的传播情况。禽流感被许多人认为是对世界公共卫生的下一个潜在最大威胁。