Interferon mediated antiviral activity against salmonid fish viruses in BF-2 and other cell lines

Abstract

Double-stranded RNA and type I interferon-like activity induce an antiviral state in vertebrate cells and in several fish cell lines by increasing the expression of proteins that inhibit virus replication. We compared the protection induced by the polyinosinic:polycytidylic acid (poly I:C) or poly I:C plus transfection agents against the infectious pancreatic necrosis virus (IPNV) and the infectious hematopoietic necrosis virus (IHNV) in BF-2 cells, with that induced in RTG-2, CHSE-214, or SAF cells. In addition, we examined the reduction in the infective titers of these viruses and the correlation with Mx protein expression as IFN marker. Furthermore, the suitability of BF-2 cells for the evaluation and optimization of immune responses in an IPNV-IHNV co-infection was assessed. The results demonstrated strong anti-IPNV and anti-IHNV activity (around 90% of infected cells surviving) in BF-2 cells transfected with poly I:C, in which a loss of 1 log 10 or 3 log 10 of the IPNV or IHNV infective titers, respectively, was observed. No antiviral activity was evident in the cells incubated with poly I:C alone. The protection recorded in the co-infection experiments was comparable with those of the single infections. The SAF cell line exhibited the lowest antiviral capacity (45%), which was also increased after transfection with poly I:C. In addition, medium from transfected BF-2 provided protection against IPNV (1 log 10 loss of infective titer) and IHNV (2 log 10 loss of infective titer) in new monolayers, indicating that these cells secreted the factors that induce antiviral activity. A correlation between antiviral activity and Mx protein expression was observed in all the cells. These results indicate that poly I:C transfection could improve IFN-like production in these cell lines. However, the antiviral effectiveness of poly I:C differed between cell lines. On the basis of our findings, we conclude that the BF-2 cell line is a useful model in which to study the role of IFN-induced cytokines in resistance against single or double infections with salmonid fish viruses.