Graduate Thesis Or Dissertation
 

Persistent infection of salmonid cell lines with infectious pancreatic necrosis virus : a model for the carrier state in trout

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  • Persistent infections with infectious pancreatic necrosis virus (IPNV) were established in two salmonid cell lines and in live brook trout. Virus inocula containing high concentrations of defective interfering IPNV enhanced but were not a requirement for the establishment of persistent infections in cell lines derived from steelhead trout (STE-137) and chinook salmon (CHSE-214) embryos. Persistent infections were initiated in live brook trout by feeding a ration containing IPNV. Viral persistence was demonstrated by the detection of infectious virus in cell culture fluids and carrier trout feces. A fluctuating release of low levels of virus was characteristic of viral persistence both in vitro and in vivo. Infectious virus was produced by 0.06 to 1.0% of the cells in carrier brook trout tissues and cell lines persistently infected with IPNV. Viral persistence in these cell lines was further characterized by the presence of viral antigens in 1% or less of the cell population. These cells were often in advanced stages of infection indicating that death results from viral replication. The remainder of the cells in the population is protected from superinfection with homologous but not heterologous viruses. Protection is disrupted by passages of these cells in growth medium containing anti-IPNV antibody. Extended propagations with high levels of antibody cures persistent infection. Low levels of infectious virus released during IPNV persistence results from limiting the number of cells as well as the amount of infectious virus produced by persistently infected cells. Interferon, temperature sensitive and defective interfering viruses are known to control infectious virus production in certain persistently infected cell lines. Interferon and temperature sensitive virus were not detected during IPNV persistence in STE-137 and CHSE-214 cell lines in this study. Interferon was also not detected in carrier brook trout sera tested on rainbow trout (RTG -2) cells but this did not preclude its presence. The amount of infectious virus produced during viral persistence seems instead to be controlled by the production of defective interfering virus. Defective interfering virus was isolated from STE-137 cell culture persistently infected with IPNV by isopycnic centrifugation in CsC1. This virus had a density of 1.29 g/cc. Infectious virus (1.33 g/cc) was also produced by these cells. Defective interfering virus effectively suppressed the yields of infectious virus from control STE-137 cells infected with both viruses. The production of defective virus was also supported by electron microscopic examinations of persistently infected STE-137 cells. These results indicated that the production of defective interfering IPNV by persistently infected cell lines may control infectious virus replication and prevent the cytocidal effect that usually is accompanied by IPNV infection. The similarities between viral persistence in these cell lines and carrier brook trout suggest that the same virus-host cell interactions may be involved both in vitro and in vivo. These cell lines can therefore serve as invaluable models for understanding persistent IPNV infections.
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