Understanding the dsRNA-IFN-antiviral immunity axis in rainbow trout

先天免疫反应是动物抵御病毒入侵的第一道防线，提供广谱抗病毒保护。鱼类也不例外，事实上，刺激先天抗病毒免疫是开发新型抗病毒疗法以保护养殖鱼类的绝佳方法，这对加拿大的经济至关重要。拟议的研究计划旨在调查三种尚未在鱼类中确定的先天免疫反应，特别是虹鳟鱼（安大略省最常见的养殖鱼类）。这项工作的重点是病毒在复制过程中产生的双链RNA，它是先天抗病毒免疫的基石，I型干扰素（ifn）的强诱导剂。ifn刺激ifn刺激基因（isg）的产生，这些基因在细胞内积累形成抗病毒状态，这是一种使细胞对病毒感染产生折射的保护状态。这一途径被称为dsrna - ifn -抗病毒轴。本提案的第一个目标将研究如何使用最近发现的“自然”方法，即使用含有dsRNA的细胞外囊泡来刺激干扰素的产生。这种方法可能被证明是最佳的，因为它可以保护dsRNA免受降解，并可能减少dsRNA的细胞毒性作用。第二个目标将集中在了解干扰素在细胞内的作用，特别是研究两个isg， vig1和vig3的功能。这些基因之所以引起人们的兴趣，是因为维格-3本身和维格-1活性的产物3'-脱氧-3',4'-二脱氢- ctp （ddhCTP）可以被添加到细胞中，以增强其固有的抗病毒活性。因此，了解这两个isg的功能可能有助于开发新的抗病毒治疗方法，可以给鱼治疗病毒感染。第三个目标将集中在训练免疫，这是先天免疫系统的免疫记忆。抗病毒训练后的免疫力还没有得到很好的理解；然而，如果适当激活，它将允许年轻的动物被“训练”或记住病毒感染，以便它们在以后的生活中产生增强的抗病毒反应。训练免疫的力量在于它提供了广谱的疫苗策略，因为训练免疫可以保护动物免受一般病毒的侵害，而传统疫苗只能保护特定病毒株。拟议的研究计划侧重于发现dsrna - ifn -抗病毒轴内的新途径和反应，这不仅将提供基本的见解，而且将为在经济相关的鱼类中应用新的抗病毒疗法提供知识基础。

The innate immune response is the first line of defense against invading viruses in animals, and provides broad-spectrum antiviral protection. Fish are no exception, indeed stimulating innate antiviral immunity is an excellent approach to developing novel antiviral therapies to protect farmed fish, which are economically important for Canada. The proposed research program aims to investigate three innate immune responses that have yet to be identified in fish in general and rainbow trout (Ontario's most common farmed fish) specifically.  This work focuses on double-stranded (ds)RNA, produced by viruses during replication, which is a strong inducer of the keystone of innate antiviral immunity, type I interferons (IFNs). IFNs stimulate the production of IFN-stimulated genes (ISGs) that accumulate within the cell to form the antiviral state, a protective state that makes a cell refractive to virus infection. This pathway is known as the dsRNA-IFN-antiviral axis. The first aim of the present proposal will examine how to stimulate interferon production with dsRNA using a `natural' method just recently discovered, namely using extracellular vesicles containing dsRNA . This method may prove to be optimal, as it would protect dsRNA from degradation and may reduce dsRNA's cytotoxic effects. The second aim will focus on understanding interferon effects within the cell, specifically looking at the function of two ISGs, vig-1 and vig-3. These genes are of interest because vig-3 itself and the product of vig-1 activity, 3'-deoxy-3', 4'-didehydro-CTP (ddhCTP), can be added to cells to enhance their innate antiviral activity. Thus understanding the function of these two ISGs may help develop novel antiviral treatments that could be given to fish to treat virus infections. The third aim will focus on trained immunity, which is immunological memory of the innate immune system. Antiviral trained immunity is not well understood; however, if properly activated, it would allow for young animals to be `trained' or remember virus infections in order for them to mount enhanced antiviral responses later in life. The power of trained immunity is that it provides broad-spectrum vaccine strategies, as trained immunity protects an animal against viruses in general, as opposed to traditional vaccines that provide protection to a specific strain of virus. The proposed research program focuses on discovering novel pathways and responses within the dsRNA-IFN-antiviral axis that will not only provide fundamental insights but the knowledge base for the application of novel antiviral therapies in an economically relevant fish species.