Influenza A Virus Infection of Human Nasal Epithelial Cells

甲型流感病毒感染人鼻上皮细胞

基本信息

批准号：
8484347
负责人：
Andrew S. Pekosz
金额：
$ 48.07万
依托单位：
JOHNS HOPKINS UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-06-11 至 2016-05-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8484347
关键词：
Affect Antiviral Agents Attention Attenuated Attenuated Live Virus Vaccine Cell Culture System Cell Culture Techniques Cells Data Defect Disease Disease Outcome Effectiveness Elderly Engineering Epithelial Cells Evaluation Genes Genetic Goals Human Immune Immune Response Genes Immune response Immunity In Vitro Inactivated Vaccines Infection Influenza Influenza A Virus, H1N1 Subtype Influenza A virus Intramuscular Kinetics Life Mediating Morbidity - disease rate Morphology Mutation Nose Population Populations at Risk Production Proteins Public Health Relative (related person)Respiratory System Respiratory tract structure Signal Pathway Signal Transduction System Therapeutic Ursidae Family Vaccination Vaccines Viral Viral Genes Viral Genome Viral Proteins Virion Virus Virus Diseases Virus Replication anti-influenza attenuation chemokine cytokine improved influenza epidemic influenza virus strain influenza virus vaccine influenzavirus insight mortality novel pandemic disease respiratory response seasonal influenza vaccine efficacy viral RNA

项目摘要

DESCRIPTION (provided by applicant): Influenza A virus infection is an important cause of annual morbidity and mortality in the human population. Both inactivated and live attenuated vaccines are available but their effectiveness could be improved substantially to provide broader and longer lasting immunity. We have established a system for isolating and differentiating primary human nasal epithelial cell (hNECs) cultures with the goal of gaining a better understanding of the viral and cellular factors that are critical for virus replication. Our data demonstrate that live, attenuated influenza vaccine strains (LAIV) show a much greater degree of attenuation in hNEC cultures than they display in other cell culture systems, suggesting there are nasal epithelial cell specific factors contributing to the attenuated replication that have yetto be identified. This attenuation appears to be at the level of producing infectious virus particles because there appears to be little difference in secreted viral proteins but a 10,000 fold reduction in infectious virus production after LAIV infection of hNEC cultures. In Aim1 we will assess the innate immune factors that are differentially induced after infection of hNECs with wild type or LAIV strains of influenza to identify cellular factors that might be mediating this difference. In Aim 2 we will focus on identifying the antiviral signaling pathways and cytokines/chemokines that are differentially induced. In both Aims we will not only identify factors but use a variety of genetic and protein manipulations to demonstrate the relative contribution of each factor to LAIV attenuation. Finally, in Aim 3 we will characterize the defect in infectious virus production and determine what viral gene segments and mutations are contributing to the hNEC specific defect in LAIV infectious virus particle production. Our data wil allow us to identify novel cellular and viral factors that will provide targets for influenza virus therapeutics and that can be used to adjust the replication of LAIV in a manner that will improve its effectiveness as a vaccine.

描述（由申请人提供）：流感病毒感染是人口年度发病率和死亡率的重要原因。可灭活和活衰减的疫苗都可以使用，但可以大大提高其有效性，以提供更广泛，更持久的免疫力。我们已经建立了一个系统，用于隔离和区分原发性人鼻上皮细胞（HNEC）培养物，目的是更好地了解对病毒复制至关重要的病毒和细胞因子。我们的数据表明，与其他细胞培养系统相比，HNEC培养物中活性，减弱的流感疫苗菌株（LAIV）表明，在HNEC培养物中的衰减程度要大得多，这表明存在鼻中皮细胞的特异性因子，这些因素有助于减弱的复制，尚待鉴定出来。这种衰减似乎处于产生传染性病毒颗粒的水平，因为分泌的病毒蛋白似乎没有差异，但是在HNEC培养物的LAIV感染后，感染性病毒产生的降低了10,000倍。在AIM1中，我们将评估具有野生型或LAIV菌株感染流感的HNEC后差异诱导的先天免疫因子，以鉴定可能介导这种差异的细胞因子。在AIM 2中，我们将专注于鉴定差异诱导的抗病毒信号通路和细胞因子/趋化因子。在这两个目标中，我们都将不仅识别因素，还可以使用各种遗传和蛋白质操纵来证明每个因素对LAIV衰减的相对贡献。最后，在AIM 3中，我们将表征传染性病毒产生的缺陷，并确定哪些病毒基因片段和突变有助于LAIV传染性病毒颗粒产生的HNEC特异性缺陷。我们的数据将使我们能够鉴定出新型的细胞和病毒因素，这些因素将为流感病毒提供靶标可以用来以提高其作为疫苗有效性的方式来调整LAIV的复制。