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Ribavirin Inhibits Parrot Bornavirus 4 Replication in Cell Culture.

Musser JM, Heatley JJ, Koinis AV, Suchodolski PF, Guo J, Escandon P, Tizard IR - PLoS ONE (2015)

Bottom Line: We tested the ability of ribavirin, an antiviral nucleoside analog with antiviral activity against a range of RNA and DNA viruses, to inhibit parrot bornavirus 4 replication in duck embryonic fibroblast cells.The addition of guanosine diminished the antiviral activity of ribavirin suggesting that one possible mechanism of action against parrot bornavirus 4 may likely be through inosine monophosphate dehydrogenase inhibition.This study demonstrates parrot bornavirus 4 susceptibility to ribavirin in cell culture.

View Article: PubMed Central - PubMed

Affiliation: Schubot Exotic Bird Health Center, College of Veterinary Medicine, Texas A&M University, College Station, Texas, United States of America; Department of Veterinary Pathobiology, College of Veterinary Medicine, Texas A&M University, College Station, Texas, United States of America.

ABSTRACT
Parrot bornavirus 4 is an etiological agent of proventricular dilatation disease, a fatal neurologic and gastrointestinal disease of psittacines and other birds. We tested the ability of ribavirin, an antiviral nucleoside analog with antiviral activity against a range of RNA and DNA viruses, to inhibit parrot bornavirus 4 replication in duck embryonic fibroblast cells. Two analytical methods that evaluate different products of viral replication, indirect immunocytochemistry for viral specific nucleoprotein and qRT-PCR for viral specific phosphoprotein gene mRNA, were used. Ribavirin at concentrations between 2.5 and 25 μg/mL inhibited parrot bornavirus 4 replication, decreasing viral mRNA and viral protein load, in infected duck embryonic fibroblast cells. The addition of guanosine diminished the antiviral activity of ribavirin suggesting that one possible mechanism of action against parrot bornavirus 4 may likely be through inosine monophosphate dehydrogenase inhibition. This study demonstrates parrot bornavirus 4 susceptibility to ribavirin in cell culture.

No MeSH data available.


Related in: MedlinePlus

Digital image analysis to determine stained PaBV-4 positive cell foci and area of infected cells.(A) Photograph after indirect immunocytochemistry of wells containing infected duck embryonic fibroblasts infected with PaBV-4 and incubated with or without ribavirin. PaBV-4 nucleoprotein detected as dense shades of grey. (B) Digital 8-bit grey-scale images of the same wells after digital scanning. (C) The same wells after 8-bit grey scale images were enhanced and converted to a binary image for analyses using ImageJ Analyze. The areas of N protein immunocomplex are in black. The area of interest that was analyzed is the same for each well and is within the superimposed yellow circles.
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pone.0134080.g001: Digital image analysis to determine stained PaBV-4 positive cell foci and area of infected cells.(A) Photograph after indirect immunocytochemistry of wells containing infected duck embryonic fibroblasts infected with PaBV-4 and incubated with or without ribavirin. PaBV-4 nucleoprotein detected as dense shades of grey. (B) Digital 8-bit grey-scale images of the same wells after digital scanning. (C) The same wells after 8-bit grey scale images were enhanced and converted to a binary image for analyses using ImageJ Analyze. The areas of N protein immunocomplex are in black. The area of interest that was analyzed is the same for each well and is within the superimposed yellow circles.

Mentions: An equally sized area of interest, maximized to include as much of the well as possible, was delineated in each well. The number of PaBv-4 positive cell foci per area of interest was counted using the ImageJ Analyze Particles tool, with particle size gated between 0.0001 to infinity square inch. The area of infected cells, measured as the percent area of black pixels per area of interest, was determined using the macro function Calculate_Black_to_White_Ratio. A representation of the wells and images is provided in Fig 1.


Ribavirin Inhibits Parrot Bornavirus 4 Replication in Cell Culture.

Musser JM, Heatley JJ, Koinis AV, Suchodolski PF, Guo J, Escandon P, Tizard IR - PLoS ONE (2015)

Digital image analysis to determine stained PaBV-4 positive cell foci and area of infected cells.(A) Photograph after indirect immunocytochemistry of wells containing infected duck embryonic fibroblasts infected with PaBV-4 and incubated with or without ribavirin. PaBV-4 nucleoprotein detected as dense shades of grey. (B) Digital 8-bit grey-scale images of the same wells after digital scanning. (C) The same wells after 8-bit grey scale images were enhanced and converted to a binary image for analyses using ImageJ Analyze. The areas of N protein immunocomplex are in black. The area of interest that was analyzed is the same for each well and is within the superimposed yellow circles.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4519282&req=5

pone.0134080.g001: Digital image analysis to determine stained PaBV-4 positive cell foci and area of infected cells.(A) Photograph after indirect immunocytochemistry of wells containing infected duck embryonic fibroblasts infected with PaBV-4 and incubated with or without ribavirin. PaBV-4 nucleoprotein detected as dense shades of grey. (B) Digital 8-bit grey-scale images of the same wells after digital scanning. (C) The same wells after 8-bit grey scale images were enhanced and converted to a binary image for analyses using ImageJ Analyze. The areas of N protein immunocomplex are in black. The area of interest that was analyzed is the same for each well and is within the superimposed yellow circles.
Mentions: An equally sized area of interest, maximized to include as much of the well as possible, was delineated in each well. The number of PaBv-4 positive cell foci per area of interest was counted using the ImageJ Analyze Particles tool, with particle size gated between 0.0001 to infinity square inch. The area of infected cells, measured as the percent area of black pixels per area of interest, was determined using the macro function Calculate_Black_to_White_Ratio. A representation of the wells and images is provided in Fig 1.

Bottom Line: We tested the ability of ribavirin, an antiviral nucleoside analog with antiviral activity against a range of RNA and DNA viruses, to inhibit parrot bornavirus 4 replication in duck embryonic fibroblast cells.The addition of guanosine diminished the antiviral activity of ribavirin suggesting that one possible mechanism of action against parrot bornavirus 4 may likely be through inosine monophosphate dehydrogenase inhibition.This study demonstrates parrot bornavirus 4 susceptibility to ribavirin in cell culture.

View Article: PubMed Central - PubMed

Affiliation: Schubot Exotic Bird Health Center, College of Veterinary Medicine, Texas A&M University, College Station, Texas, United States of America; Department of Veterinary Pathobiology, College of Veterinary Medicine, Texas A&M University, College Station, Texas, United States of America.

ABSTRACT
Parrot bornavirus 4 is an etiological agent of proventricular dilatation disease, a fatal neurologic and gastrointestinal disease of psittacines and other birds. We tested the ability of ribavirin, an antiviral nucleoside analog with antiviral activity against a range of RNA and DNA viruses, to inhibit parrot bornavirus 4 replication in duck embryonic fibroblast cells. Two analytical methods that evaluate different products of viral replication, indirect immunocytochemistry for viral specific nucleoprotein and qRT-PCR for viral specific phosphoprotein gene mRNA, were used. Ribavirin at concentrations between 2.5 and 25 μg/mL inhibited parrot bornavirus 4 replication, decreasing viral mRNA and viral protein load, in infected duck embryonic fibroblast cells. The addition of guanosine diminished the antiviral activity of ribavirin suggesting that one possible mechanism of action against parrot bornavirus 4 may likely be through inosine monophosphate dehydrogenase inhibition. This study demonstrates parrot bornavirus 4 susceptibility to ribavirin in cell culture.

No MeSH data available.


Related in: MedlinePlus