ENDOTHELIAL CELL TROPISM IN THE PATHOGENESIS AND HOST RESPONSE AGAINST INFLUENZA VIRUSES - RESUBMISSION 01

发病机制中的内皮细胞趋向性和宿主对流感病毒的反应 - 重新提交 01

• 批准号: 9769437
• 负责人: Balaji Manicassamy
• 金额: $ 55.63万
• 依托单位: UNIVERSITY OF IOWA
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-12-08 至 2022-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9769437
• 关键词: Adult; Adult Respiratory Distress Syndrome; Antiviral Response; Attenuated Live Virus Vaccine; Avian Influenza A Virus; Biological Models; Blood Vessels; Cell Adhesion Molecules; Cells; Characteristics; Coculture Techniques; Development; Disease; Disease Progression; Down-Regulation; Elderly; Endothelial Cells; Endothelium; Engineering; Epidemic; Epithelial Cells; Extravasation; Ferrets; Generations; Goals; H7N7; Health; Hematopoietic; Human; Immune; Immune response; Immunocompromised Host; Individual; Infection; Inflammatory; Inflammatory Response; Influenza; Influenza A Virus, H1N1 Subtype; Influenza A Virus, H5N1 Subtype; Influenza A Virus, H7N9 Subtype; Influenza A virus; Injury; Innate Immune Response; Integration Host Factors; Knowledge; Link; Lower respiratory tract structure; Lung; Mediating; MicroRNAs; Modeling; Molecular; Morbidity - disease rate; Mus; Nature; Neutrophil Infiltration; Nose; Pathogenesis; Pathogenicity; Positioning Attribute; Predisposition; Production; Pulmonary Edema; Reporter; Reporting; Research Personnel; Role; Severity of illness; Site; Survival Rate; Symptoms; Testing; Therapeutic; Transgenic Organisms; Tropism; Upper Respiratory Infections; Vascular Permeabilities; Viral; Viral Genes; Viral Genome; Viral Load result; Viral Pathogenesis; Viral Pneumonia; Virulence; Virulent; Virus; Virus Diseases; Virus Replication; Vulnerable Populations; age group; burden of illness; cell type; cytokine; design; experimental study; immunopathology; improved; in vivo; influenza virus strain; influenzavirus; lung injury; member; monocyte; mortality; mouse model; neutrophil; novel; novel therapeutics; pandemic disease; pathogen; prevent; recombinant virus; recruit; seasonal influenza; tissue tropism; tool

ABSTRACT Influenza A viruses pose a serious threat to human health, causing seasonal epidemics and occasional pandemics that result in significant morbidity and mortality worldwide. The virulence and pathogenic nature of influenza virus strains vary greatly, and is often determined by the constellation of viral genes. While seasonal influenza viruses cause a mild upper respiratory infection in healthy individuals, highly pathogenic avian influenza viruses (HPAIV) of the H5N1 subtype replicate in the lower respiratory tract, causing fatal viral pneumonia characterized by pulmonary edema and vascular leakage. In addition, H5N1 infections are associated with an uncontrolled activation of host immune responses, which contributes to the severity of disease. The role of viral and host factors in the enhanced virulence and pulmonary immunopathology characteristic of HPAIV infections remain unknown. To this end, we engineered H5N1 viruses with restricted tropism through the incorporation of microRNA (miRNA) target sites into the viral genome. Specifically, we generated an H5N1 virus carrying endothelial cell specific miR-126 target sites (H5N1-126T), such that viral replication was abrogated in endothelial cells without impeding replication in other cell types. In our recent studies, H5N1-126T infected mice and ferrets showed significantly reduced virulence and vascular leakage in the lungs as compared to the control H5N1-ScrbT virus infected group, despite similar viral loads in the lungs of mice and nasal washes of ferrets. To our knowledge, this is the first study to demonstrate the importance of endothelial cell tropism to H5N1 pathogenesis. The current proposal will (1) determine the consequence of endothelial cell infection to barrier integrity and function, (2) investigate the role of endothelial cell tropism in orchestrating immune responses, and (3) assess the contribution of viral tropism to the pathogenesis of other highly virulent strains including H7N7, 1918 H1N1, and H7N9 in mice and ferrets. The knowledge gained from these studies will help to elucidate the mechanism by which virulent influenza viruses cause severe disease and aid in the development of novel therapeutic strategies against virulent influenza strains. Furthermore, these studies will reveal how viral tropism modulates host antiviral responses and will pave the way for the rational design of novel live attenuated vaccines with restricted tropism.

摘要 甲型流感病毒对人类健康构成严重威胁，造成季节性流行， 在全世界造成严重发病率和死亡率的流行病。的毒力和致病性 流感病毒株的差异很大，并且通常由病毒基因群决定。虽然季节性 流感病毒在健康个体、高致病性禽流感病毒、高致病性禽流感病毒和高致病性禽流感病毒中引起轻度上呼吸道感染。 H5 N1亚型的流感病毒（HPAIV）在下呼吸道复制，引起致命的病毒感染。 以肺水肿和血管渗漏为特征的肺炎。此外，H5 N1感染是 与宿主免疫反应的不受控制的激活有关，这有助于严重的 疾病病毒和宿主因子在增强毒力和肺免疫病理学中的作用 HPAIV感染的特征仍然未知。为此，我们设计了H5 N1病毒， 通过将微小RNA（miRNA）靶位点掺入病毒基因组中来增强嗜性。我们特别 产生了携带内皮细胞特异性miR-126靶位点的H5 N1病毒（H5 N1 - 126 T），使得病毒 内皮细胞中的复制被消除，而不妨碍其他细胞类型中的复制。在我们最近 研究表明，H5 N1 - 126 T感染的小鼠和雪貂的毒力和血管渗漏显著降低， 与对照H5 N1-ScrbT病毒感染组相比， 老鼠和雪貂的鼻洗液。据我们所知，这是第一项证明 内皮细胞对H5 N1致病性的嗜性。目前的建议将（1）确定以下后果： 内皮细胞感染对屏障完整性和功能的影响，（2）研究内皮细胞嗜性在 协调免疫反应，和（3）评估病毒嗜性对其他疾病发病机制的贡献。 在小鼠和雪貂中的高毒力毒株包括H7 N7、1918 H1N1和H7N9。知识来自于 这些研究将有助于阐明强毒流感病毒引起严重疾病的机制 并有助于开发针对毒性流感毒株的新治疗策略。此外，委员会认为， 这些研究将揭示病毒的嗜性如何调节宿主的抗病毒反应， 合理设计具有限制性嗜性的新型减毒活疫苗。