6-methylmercaptopurine riboside, a thiopurine nucleoside with antiviral activity against canine distemper virus in vitro

Propolis is a resin that honey bees (Apis mellifera) produce by mixing wax, exudates collected from tree shoots, pollen, and enzymes. It has been used for its biological properties against pathogenic microorganisms including those of viral origin. In the present study, we demonstrate the antiviral effect of Mexican propolis, as well as of the three commercial flavonoids (quercetin, naringenin, and pinocembrin) present in its composition, in cell cultures infected with Canine Distemper Virus. The treatments were carried out with propolis, flavonoids individually, and a mixture of the three flavonoids at three different times. Antiviral activity was evaluated by the inhibition of the relative expression of the virus nucleoprotein gene (Real-Time qPCR) and by the determination of cellular viability (MTT assay). Propolis applied before infection decreased viral expression (0.72 versus 1.0, 1.65, and 1.75 relative expressions) and correlated with increased cell viability (0.314 versus 0.215, 0.259, and 0.237 absorbance units (AU)). The administration of a flavonoid mixture containing the three commercial flavonoids before infection induces a slight decrease in viral expression (0.93 versus 1, 1.42, and 1.82 relative expressions); however, it does not improve cellular viability (0.255 versus 0.247, 0.282, and 0.245 AU). Quercetin administrated at the same time of infection decreases viral expression (0.90 versus 1.0, 3.25, and 1.02 relative expressions) and improves cellular viability (0.294 versus 0.240, 0.250, and 0.245 AU). Pinocembrin and naringenin individually did not show any antiviral activity at the administration times evaluated in this study. The present work is the first in vitro study of the effect of propolis in Canine Distemper Virus and demonstrated the antiviral activity of Mexican propolis, in addition to the synergy that exists between the three flavonoids on cell viability and the expression of the nucleoprotein virus gene.

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In Vitro Evaluation of Antiviral Activity of Ribavirin Against Canine Distemper Virus

Veterinary Research Communications ◽

10.1007/s11259-006-0058-0 ◽

2006 ◽

Vol 30 (S1) ◽

pp. 269-272 ◽

Cited By ~ 8

Author(s):

A. Scagliarini ◽

F. Vaccari ◽

L. Gallina ◽

F. Dal Pozzo ◽

S. Prosperi

Keyword(s):

Antiviral Activity ◽

Canine Distemper Virus ◽

Canine Distemper ◽

In Vitro Evaluation

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Virulence of tissue culture-propagated canine distemper virus

Infection and Immunity ◽

10.1128/iai.29.3.940-944.1980 ◽

1980 ◽

Vol 29 (3) ◽

pp. 940-944 ◽

Cited By ~ 1

Author(s):

A E Metzler ◽

R J Higgins ◽

S Krakowka ◽

A Koestner

Keyword(s):

Tissue Culture ◽

Neurological Disease ◽

Canine Distemper Virus ◽

In Vitro Growth ◽

Canine Distemper ◽

Passage Level ◽

Pulmonary Macrophage

Virulence of canine distemper virus (CDV) adapted to in vitro growth in Vero or bovine cells was determined by inoculation into CDV-susceptible neonatal gnotobiotic dogs. When compared with dogs given virulent R252-CDV, Vero R252-CDV was attenuated at passage level 14. In contrast, dogs inoculated with bovine R252-CDV at the same passage level experienced rapid fatal neurological disease. Virulence was not linked to ability to infect or replicate in canine pulmonary macrophage cultures. Retention of virulence by bovine R252-CDV is unique and worthy of further study.

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Virulent and attenuated canine distemper virus infects multiple dog brain cell types in vitro

Glia ◽

10.1002/glia.440040409 ◽

1991 ◽

Vol 4 (4) ◽

pp. 408-416 ◽

Cited By ~ 14

Author(s):

Susan Pearce-Kelling ◽

William J. Mitchell ◽

Brian A. Summers ◽

Max J. G. Appel

Keyword(s):

Canine Distemper Virus ◽

Cell Types ◽

Brain Cell ◽

Canine Distemper ◽

Dog Brain ◽

Brain Cell Types

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Canine distemper virus-induced glial cell changes in vitro

Acta Neuropathologica ◽

10.1007/bf00684920 ◽

1983 ◽

Vol 62 (1-2) ◽

pp. 51-58 ◽

Cited By ~ 12

Author(s):

A. Zurbriggen ◽

M. Vandevelde

Keyword(s):

Glial Cell ◽

Canine Distemper Virus ◽

Canine Distemper ◽

Cell Changes

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Complement-Mediated Neutralization of Canine Distemper Virus In Vitro: Cross-Reaction between Vaccine Onderstepoort and Field KDK-1 Strains with Different Hemagglutinin Gene Characteristics

Clinical and Vaccine Immunology ◽

10.1128/cdli.9.4.921-924.2002 ◽

2002 ◽

Vol 9 (4) ◽

pp. 921-924 ◽

Cited By ~ 2

Author(s):

Masami Mochizuki ◽

Megumi Motoyoshi ◽

Ken Maeda ◽

Kazunari Kai

Keyword(s):

Guinea Pig ◽

Canine Distemper Virus ◽

Field Isolate ◽

Neutralization Assay ◽

Cross Reaction ◽

Canine Distemper ◽

Hemagglutinin Gene

ABSTRACT The properties of neutralization of antigens of canine distemper virus Onderstepoort and a recent field isolate, KDK-1, were investigated with strain-specific dog sera. A conventional neutralization assay indicated antigenic dissimilarity between the strains; however, when guinea pig complement was included in the reaction mixture, the strains were neutralized with not only the homologous but also the heterologous antibodies.

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Inhibition of canine distemper virus replication by blocking pyrimidine nucleotide synthesis with A77 1726, the active metabolite of the anti-inflammatory drug leflunomide

Journal of General Virology ◽

10.1099/jgv.0.001534 ◽

2021 ◽

Author(s):

Yao Li ◽

Li Yi ◽

Sipeng Cheng ◽

Yongshan Wang ◽

Jiongjiong Wang ◽

...

Keyword(s):

Antiviral Activity ◽

Active Metabolite ◽

Canine Distemper Virus ◽

De Novo ◽

Specific Inhibitor ◽

Vero Cells ◽

Canine Distemper ◽

Nucleotide Synthesis ◽

Pyrimidine Nucleotide ◽

Rate Limiting

Canine distemper virus (CDV) is the aetiological agent that causes canine distemper (CD). Currently, no antiviral drugs have been approved for CD treatment. A77 1726 is the active metabolite of the anti-rheumatoid arthritis (RA) drug leflunomide. It inhibits the activity of Janus kinases (JAKs) and dihydroorotate dehydrogenase (DHO-DHase), a rate-limiting enzyme in de novo pyrimidine nucleotide synthesis. A77 1726 also inhibits the activity of p70 S6 kinase (S6K1), a serine/threonine kinase that phosphorylates and activates carbamoyl-phosphate synthetase (CAD), a second rate-limiting enzyme in the de novo pathway of pyrimidine nucleotide synthesis. Our present study focuses on the ability of A77 1726 to inhibit CDV replication and its underlying mechanisms. Here we report that A77 1726 decreased the levels of the N and M proteins of CDV and lowered the virus titres in the conditioned media of CDV-infected Vero cells. CDV replication was not inhibited by Ruxolitinib (Rux), a JAK-specific inhibitor, but by brequinar sodium (BQR), a DHO-DHase-specific inhibitor, and PF-4708671, an S6K1-specific inhibitor. Addition of exogenous uridine, which restores intracellular pyrimidine nucleotide levels, blocked the antiviral activity of A77 1726, BQR and PF-4708671. A77 1726 and PF-4708671 inhibited the activity of S6K1 in CDV-infected Vero cells, as evidenced by the decreased levels of CAD and S6 phosphorylation. S6K1 knockdown suppressed CDV replication and enhanced the antiviral activity of A77 1726. These observations collectively suggest that the antiviral activity of A77 1726 against CDV is mediated by targeting pyrimidine nucleotide synthesis via inhibiting DHO-DHase activity and S6K1-mediated CAD activation.

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Recovery of NanoLuc Luciferase-Tagged Canine Distemper Virus for Facilitating Rapid Screening of Antivirals in vitro

Frontiers in Veterinary Science ◽

10.3389/fvets.2020.600796 ◽

2020 ◽

Vol 7 ◽

Author(s):

Fuxiao Liu ◽

Qianqian Wang ◽

Yilan Huang ◽

Ning Wang ◽

Youming Zhang ◽

...

Keyword(s):

High Throughput ◽

High Throughput Screening ◽

Canine Distemper Virus ◽

Reverse Genetics ◽

Rapid Screening ◽

Canine Distemper ◽

Virus Propagation ◽

The Family ◽

Conventional Methods

Canine distemper virus (CDV), belonging to the genus Morbillivirus in the family Paramyxoviridae, is a highly contagious pathogen, affecting various domestic, and wild carnivores. Conventional methods are too cumbersome to be used for high-throughput screening of anti-CDV drugs. In this study, a recombinant CDV was rescued using reverse genetics for facilitating screening of anti-CDV drug in vitro. The recombinant CDV could stably express the NanoLuc® luciferase (NLuc), a novel enzyme that was smaller and “brighter” than others. The intensity of NLuc-catalyzed luminescence reaction indirectly reflected the anti-CDV effect of a certain drug, due to a positive correlation between NLuc expression and virus propagation in vitro. Based on such a characteristic feature, the recombinant CDV was used for anti-CDV assays on four drugs (ribavirin, moroxydine hydrochloride, 1-adamantylamine hydrochloride, and tea polyphenol) via analysis of luciferase activity, instead of via conventional methods. The result showed that out of these four drugs, only the ribavirin exhibited a detectable anti-CDV effect. The NLuc-tagged CDV would be a rapid tool for high-throughput screening of anti-CDV drugs.

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