Early Responses to Influenza A Virus Replication In Vivo

对甲型流感病毒体内复制的早期反应

• 批准号: 10579883
• 负责人: Ryan Langlois
• 金额: $ 50.13万
• 依托单位: UNIVERSITY OF MINNESOTA
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-03-01 至 2025-02-28
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10579883
• 关键词: Alveolar Cell; Animals; Antiviral Response; Autocrine Communication; Cell Shape; Cells; Cellular Tropism; Chromatin; Complex; Cytoprotection; Data; Environment; Epigenetic Process; Epithelial Cells; Event; Flow Cytometry; Fluorescence; Gene Cluster; Genes; Genetic Engineering; Goals; Hemagglutinin; Heterogeneity; Human; Immune response; In Vitro; Infection; Influenza; Influenza A virus; Innate Immune Response; Integration Host Factors; Interferons; Knowledge; Label; Laboratories; Life Cycle Stages; Lung; Microscopy; Mus; Natural Immunity; Paracrine Communication; Positioning Attribute; Process; Proteins; RNA Interference; Reporter; Research; Respiratory System; Role; Seasons; Secondary to; Sentinel; Series; Shapes; Supporting Cell; System; Techniques; Testing; Therapeutic; Time; Tropism; Vaccines; Viral; Viral Genes; Virus; Virus Diseases; Virus Replication; Work; antiviral immunity; cell type; design; experimental study; fluorophore; in vivo; influenza infection; influenzavirus; innovation; mutant; pandemic disease; pathogen; receptor; response; tool; viral detection; virus host interaction; virus tropism

Project Summary Influenza A virus is a seasonal pathogen with the potential to unpredictably cause devastating pandemics. Influenza has a broad tropism within the respiratory tract infecting many different subsets of epithelial cells. These epithelial cells are the primary target of influenza infection and are responsible for amplifying and spreading the infection. These cells are also critical sentinels detecting the virus and initiating antiviral immune responses. It is difficult to determine early replication events because it is hard to label infected cells rapidly after infection. Additionally, the virus spreads within the lung making it difficult to disentangle low replication levels from new infections. To overcome these caveats, we used a single cycle reporter system that robustly labels infected cells and is restricted to the first cell types infected. Our preliminary data demonstrate that cells supporting different levels of virus replication express distinct sets of interferon-stimulated genes. These data demonstrate that the antiviral response is tuned to level of replication. Using a sequential infection strategy where the single cycle viruses express discrete fluorophores to specifically label primary and secondary infected cells we have demonstrated that ciliated epithelial cells are specifically protected during the second wave of virus replication. These data demonstrate that virus tropism is significantly shaped by innate immune responses during virus dissemination. The objective of this research is to determine responses that inhibit influenza virus replication and to determine how virus tropism is altered during innate immune responses in vivo. Our central hypothesis is that cellular responses to infection become tailored to the stages of viral replication and the round of infection which impacts overall viral replication levels and cellular tropism. Our studies will address this hypothesis through use of a combination of reporter viruses that can determine the degree, stage, and round of replication to elucidate fundamental antiviral processes in vivo. We will address this hypothesis in two aims. Aim 1 will be to determine how varying levels of replication induce unique ISGs and the role of the resulting effector proteins in the antiviral response. Aim 2 will focus on elucidating the mechanisms of protection of epithelial cell subsets during virus spread. Results from this proposal will uncover fundamental mechanisms in virus-host interactions and antiviral immunity.

项目概要 甲型流感病毒是一种季节性病原体，有可能不可预测地引起毁灭性的流行病。 流感在呼吸道内具有广泛的趋向性，可感染许多不同的上皮细胞亚群。 这些上皮细胞是流感感染的主要目标，负责扩增和 传播感染。这些细胞也是检测病毒和启动抗病毒免疫的关键哨兵 回应。由于很难快速标记受感染的细胞，因此很难确定早期复制事件 感染后。此外，病毒在肺部传播，因此很难解决低复制问题 新感染的水平。为了克服这些问题，我们使用了单循环报告系统，该系统可以稳健地 标记受感染的细胞并仅限于第一个受感染的细胞类型。我们的初步数据表明，细胞 支持不同水平的病毒复制，表达不同的干扰素刺激基因组。这些数据 证明抗病毒反应根据复制水平进行调节。使用顺序感染策略 其中单循环病毒表达离散荧光团以特异性标记初级和次级 我们已经证明，纤毛上皮细胞在感染细胞的第二次感染过程中受到特别保护。 病毒复制波。这些数据表明，病毒的趋向性很大程度上受到先天免疫的影响 病毒传播过程中的反应。这项研究的目的是确定抑制反应 流感病毒复制并确定病毒向性在先天免疫反应过程中如何改变 体内。我们的中心假设是，细胞对感染的反应是根据病毒的阶段而定制的 复制和感染轮次影响整体病毒复制水平和细胞向性。我们的 研究将通过使用报告病毒的组合来解决这一假设，这些报告病毒可以确定 复制的程度、阶段和轮次，以阐明体内基本的抗病毒过程。我们将解决 这个假设有两个目的。目标 1 是确定不同的复制水平如何诱导独特的 ISG 以及由此产生的效应蛋白在抗病毒反应中的作用。目标 2 将重点阐明 病毒传播过程中上皮细胞亚群的保护机制。该提案的结果将揭示 病毒与宿主相互作用和抗病毒免疫的基本机制。