Department of Biology, University of Waterloo, Waterloo, ON, Canada.
Department of Pathobiology, Ontario Veterinary College, University of Guelph, Guelph, ON, Canada.
Fish Shellfish Immunol. 2017 Nov;70:214-227. doi: 10.1016/j.fsi.2017.09.001. Epub 2017 Sep 4.
Heart diseases caused by viruses are major causes of Atlantic salmon aquaculture loss. Two Atlantic salmon cardiovascular cell lines, an endothelial cell line (ASHe) from the heart and a fibroblast cell line (BAASf) from the bulbus arteriosus, were evaluated for their response to four fish viruses, CSV, IPNV, VHSV IVa and VHSV IVb, and the innate immune agonist, double-stranded RNA mimic poly IC. All four viruses caused cytopathic effects in ASHe and BAASf. However, ASHe was more susceptible to all four viruses than BAASf. When comparing between the viruses, ASHe cells were found to be moderately susceptible to CSV and VHSV IVb, but highly susceptible to IPNV and VHSV IVa induced cell death. All four viruses were capable of propagating in the ASHe cell line, leading to increases in virus titre over time. In BAASf, CSV and IPNV produced more than one log increase in titre from initial infection, but VHSV IVb and IVa did not. When looking at the antiviral response of both cell lines, Mx proteins were induced in ASHe and BAASf by poly IC. All four viruses induced Mx proteins in BAASf, while only CSV and VHSV IVb induced Mx proteins in ASHe. IPNV and VHSV IVa suppressed Mx proteins expression in ASHe. Pretreatment of ASHe with poly IC to allow for Mx proteins accumulation protected the culture from subsequent infections with IPNV and VHSV IVa, resulting in delayed cell death, reduced virus titres and reduced viral proteins expression. These data suggest that endothelial cells potentially can serve as points of infections for viruses in the heart and that two of the four viruses, IPNV and VHSV IVa, have mechanisms to avoid or downregulate antiviral responses in ASHe cells. Furthermore, the high susceptibility of the ASHe cell line to IPNV and VHSV IVa can make it a useful tool for studying antiviral compounds against these viruses and for general detection of fish viruses.
病毒引起的心脏病是大西洋鲑鱼养殖损失的主要原因。评估了两种大西洋鲑鱼心血管细胞系,一种来自心脏的内皮细胞系(ASHe)和一种来自动脉球的成纤维细胞系(BAASf),以评估它们对四种鱼类病毒、CSV、IPNV、VHSV IVa 和 VHSV IVb 以及先天免疫激动剂双链 RNA 模拟物 poly IC 的反应。四种病毒均在 ASHe 和 BAASf 中引起细胞病变效应。然而,ASHe 对所有四种病毒的敏感性均高于 BAASf。在比较病毒时,发现 ASHe 细胞对 CSV 和 VHSV IVb 中度敏感,但对 IPNV 和 VHSV IVa 诱导的细胞死亡高度敏感。所有四种病毒均能够在 ASHe 细胞系中繁殖,导致病毒滴度随时间增加。在 BAASf 中,CSV 和 IPNV 的初始感染后病毒滴度增加了一个以上对数级,但 VHSV IVb 和 IVa 则没有。当观察两种细胞系的抗病毒反应时,poly IC 诱导了 ASHe 和 BAASf 中的 Mx 蛋白。所有四种病毒均诱导了 BAASf 中的 Mx 蛋白,而只有 CSV 和 VHSV IVb 诱导了 ASHe 中的 Mx 蛋白。IPNV 和 VHSV IVa 抑制了 ASHe 中 Mx 蛋白的表达。用 poly IC 预处理 ASHe 以允许 Mx 蛋白积累可保护培养物免受随后的 IPNV 和 VHSV IVa 感染,从而导致细胞死亡延迟、病毒滴度降低和病毒蛋白表达减少。这些数据表明,内皮细胞可能是心脏中病毒感染的部位,四种病毒中的两种,即 IPNV 和 VHSV IVa,具有避免或下调 ASHe 细胞中抗病毒反应的机制。此外,ASHe 细胞系对 IPNV 和 VHSV IVa 的高敏感性使其成为研究针对这些病毒的抗病毒化合物的有用工具,并且可用于一般鱼类病毒的检测。