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

先天免疫反应是动物抵抗入侵病毒的第一道防线，并提供广谱抗病毒保护。鱼类也不例外，事实上，刺激先天性抗病毒免疫是开发新的抗病毒疗法以保护养殖鱼类的一种很好的方法，这对加拿大来说具有重要的经济意义。拟议的研究计划旨在调查三种先天免疫反应，这些反应尚未在一般鱼类和虹鳟鱼（安大略最常见的养殖鱼类）中确定。 这项工作的重点是双链（ds）RNA，由病毒在复制过程中产生，这是一个强大的诱导剂的关键先天性抗病毒免疫，I型干扰素（IFN）。IFN刺激IFN刺激基因（ISG）的产生，这些基因在细胞内积累以形成抗病毒状态，这是一种使细胞抵抗病毒感染的保护性状态。该途径被称为dsRNA-IFN-抗病毒轴。本发明的第一个目的将研究如何使用最近发现的天然方法，即使用含有dsRNA的细胞外囊泡，用dsRNA刺激干扰素产生。这种方法可能被证明是最佳的，因为它将保护dsRNA免受降解，并可能降低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，特别是观察两个ISG，vig-1和vig-3的功能。这些基因是令人感兴趣的，因为vig-3本身和vig-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它们的先天抗病毒活性。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的免疫力，这是先天免疫系统的免疫记忆。抗病毒训练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接受训练或记住病毒感染，以便它们在以后的生活中增强抗病毒反应。训练免疫的力量在于它提供了广谱疫苗策略，因为训练免疫通常保护动物免受病毒的侵害，而不是传统疫苗提供对特定病毒株的保护。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型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.