Probing the contribution of viral heterogeneity to interferon induction

项目摘要 细胞固有的先天免疫途径是几乎所有病原体在免疫过程中必须克服的障碍。 才能顺利完成它们的传染性生命周期甲型流感病毒（IAV）没有什么不同， 几种不同的策略，以逃避或以其他方式抑制宿主的识别。这对于 病毒存活，作为干扰素的产生，先天反应的中心信号分子，导致 甚至IAV也易感染的高度抗病毒状态的传播。无论如何，IAV仍然在一个 感染细胞的一小部分。为了了解并潜在地操纵宿主对IAV的反应，我们 必须首先了解那些传播干扰素反应的罕见感染的性质。是否 在IAV感染中表现出的令人难以置信的遗传和行为多样性使它们易于被识别？如果是的话， 是否存在单一的免疫刺激变异模式，或者存在多个病毒失效点？ 在两年的资助期内，罗素博士将定量衡量 IAV群体的异质性影响它们通过先天免疫途径的识别。这将需要 对大量的、感染的细胞群体以及单细胞转录组学测量进行测序-这两种方法都是 这是罗素博士在杰西·布鲁姆博士的实验室里开发和试验的。超越了 通过这些测序方法实现的观察科学，罗素博士将探索新的和 先前确定的免疫刺激性IAV变异，以确定因果关系。这将包括试图 确定所有免疫刺激变异是否通过相同的、共享的机制起作用。这将是 主要通过使用反向遗传方法重现纯病毒群体中的变异来完成， 允许独立探索免疫刺激异质性的个体来源。最后，博士。 罗素将寻求将他的工作从IAV扩展到对人类重要的相关病毒，即流感B病毒。 不同病毒谱系之间的比较将为病毒与先天性 免疫力在进化过程中是静态，还是相对动态的， 可推广的治疗干预。总的来说，这项工作将产生一个强大的框架，描述 先天免疫识别这些病毒的总和，作为一个实质性的资源，以外地移动 性新

Project Summary Cell intrinsic innate immune pathways present a barrier that nearly all pathogens must overcome in order to successfully complete their infectious lifecycle. Influenza A virus (IAV) is no different, and engages several different strategies in order to evade or otherwise suppress recognition by the host. This is crucial to viral survival, as the production of interferons, the central signaling molecules of the innate response, results in the propagation of a highly antiviral state to which even IAV is susceptible. Regardless, IAV is still detected in a small subset of infected cells. In order to understand, and potentially manipulate, the host response to IAV, we must first understand the nature of those rare infections that propagate an interferon response. Does the incredible genetic and behavioral diversity exhibited in IAV infections predispose their recognition? And, if so, is there a single stereotyped pattern of immunostimulatory variation, or do multiple points of viral failure exist? Over the course of the two-year funding period, Dr. Russell will quantitatively measure how heterogeneity in IAV populations impacts their recognition by innate immune pathways. This will require sequencing bulk, infected, populations of cells as well as single-cell transcriptomic measurements – both of which have been developed and piloted by Dr. Russell while in Dr. Jesse Bloom's lab. Moving beyond the observational science enabled by these sequencing approaches, Dr. Russell will explore both newly- and previously-identified immunostimulatory IAV variation to establish causality. This will include attempts to ascertain whether all immunostimulatory variation acts through the same, shared, mechanisms. This will be primarily accomplished by recapitulating variation in pure viral populations using a reverse-genetic approach, permitting the independent exploration of individual sources of immunostimulatory heterogeneity. Lastly, Dr. Russell will seek to expand his work beyond IAV to a related virus of human importance, Influenza B virus. Comparisons across distinct viral lineages will provide vital insight into whether viral interactions with innate immunity are static over evolutionary time, or whether they are relatively dynamic and thus intractable to a generalizable therapeutic intervention. Overall, this work will generate a robust framework describing the totality of innate immune recognition of these viruses, serving as a substantial resource to the field moving forward.