Developing and optimizing cell lines to study and control emerging viral diseases in farmed Atlantic Salmon: the HSMI and PRV case

鱼类是世界上最健康的食物来源之一;然而，捕捞渔业正在下降。 在未来，水产养殖可能会提供大多数鱼类。 因此，鱼类养殖已成为一项有价值的经济活动，但维持水产养殖业具有挑战性。 一个挑战是新出现的病毒性疾病。 大西洋鲑鱼（AS）的一个例子是心脏和骨骼肌炎症（HSMI）。 病原体被认为是鱼呼肠孤病毒（PRV）。 PRV在体外仅少量繁殖，且仅在1个细胞系中繁殖，妨碍了诊断、病毒分离和疫苗制备的容易性。 因此，该提案有五个相互作用的目标。 第一个是从心脏和脾中培养产生血细胞的培养物中开发AS细胞系。 病毒需要细胞才能复制，但对于它们将在哪些细胞中复制和暴露自己是有选择性的。 PRV的两个潜在复制位点是心脏细胞和血细胞。 细胞系可以冷冻保存，为将来研究新出现的病毒性疾病提供稳定的试剂。二是筛选研究HSMI病毒的最佳细胞培养物。 这些细胞系将是新的细胞系加上来自一系列鱼类和组织的现有细胞系。 试验材料将是来自HSMI鱼的样品。 第三是改变样品制备和暴露条件，以提高细胞培养的病毒检测。第四个目标是优化一种诊断工具，该工具可专门用于本研究的目标2至5，并通常用于研究HSMI。 这是一种聚合酶链反应，用于定量培养物和鱼类中的病毒数量。 最后，将使用细胞系生产PRV，使鱼类可以接触PRV，并测试PRV和HSMI之间的因果关系，并描述疾病的发病机制。 如果PRV导致HMSI，疫苗可以被开发出来，这些疫苗可能成为我们的合作伙伴制药公司有价值的商业产品。

Fish represent one of the most healthy sources of food in the world; yet, capture fisheries are declining. In the future, aquaculture will likely supply most fish. As a result, fish farming has become a valuable economic activity, but maintaining aquaculture is challenging. One challenge is emerging viral diseases. An example for Atlantic salmon (AS) is heart and skeletal muscle inflammation (HSMI). The causative agent is thought to be piscine reovirus (PRV). PRV reproduces only marginally in vitro and only in 1 cell line, preventing easy diagnosis, virus isolation and vaccine preparation. Therefore, this proposal has 5 interacting goals. The 1st is to develop AS cell lines from hearts and blood cell producing cultures from spleens. Viruses need cells in order to replicate but are selective as to what cells they will replicate in and reveal themselves. Two potential replication sites for PRV are cardiac cells and blood cells. Cell lines can be cryopreserved, providing stable reagents to study emerging viral diseases in the future. The 2nd is to identify the best cell cultures for studying viruses of HSMI. The cell lines will be the new ones plus existing ones from a range of fish and tissues. The test material will be samples from fish with HSMI. The 3rd is varying sample preparation and exposure conditions to improve viral detection by cell cultures. The 4th goal will be to optimize a diagnostic tool that can be used specifically in this research for goals 2 to 5 and generally to study HSMI. This is the polymerase chain reaction to quantify the amount of virus in cultures and in fish. Finally, cell lines will be used to produce PRV so that fish can be exposed and the causal relationship between PRV and HSMI tested and pathogenesis of the disease described. If PRV causes HMSI, vaccines could be developed, and these could become valuable commercial products for our partner, a pharmaceutical company.