Inhibition of Influenza Virus Ribonucleic Acid Polymerase by Ribavirin Triphosphate

Eriksson, Barbro; Helgstrand, Erik; Johansson, Nils Gunnar; Larsson, Anna‐Karin; Misiorny, Alfons; Norén, J. O.; Philipson, Lennart; Stenberg, K; Stening, G.; Stridh, S.; Öberg, Bo

doi:10.1128/aac.11.6.946

Cited by 251 publications

(163 citation statements)

References 20 publications

Supporting

Mentioning

156

Contrasting

Unclassified

Order By: Relevance

“…24) Furthermore, Eriksson et al found that ribavirin triphosphate, a cellular metabolite of ribavirin, selectively inhibits influenza virus RNA-dependent RNA polymerase. 25) We examined the effect of ribavirin on influenza virus replication in chorion and amnion cells. Virus yields in culture supernatants from both cell types increased about 300-fold in the absence of ribavirin, but increased only 3-or 4-fold at a concentration of 1 mM ribavirin (Table 1).…”

Section: Resultsmentioning

confidence: 99%

Apoptosis in Cultured Human Fetal Membrane Cells Infected with Influenza Virus.

Uchide

Ohyama

Bessho³

et al. 2002

Biological & Pharmaceutical Bulletin

View full text Add to dashboard Cite

When a virus encounters a susceptible cell, the virus enters it to initiate cytocidal, persistent, latent or abortive cell infection. The influenza virus induces cytocidal infection accompanied by virus production and cell death in several types of cultures such as HeLa and Madin-Darby canine kidney (MDCK) cell lines and human peripheral blood monocytes in vitro, all of which die through the mechanism of apoptosis. 1-4)The human influenza virus usually infects the human respiratory organs. However, influenza virus was isolated from blood, [5][6][7][8] as well as extrapulmonary regions such as lymph node, spleen, liver, kidney, adrenal glands, meninges, 7,[9][10][11] and cerebrospinal fluid.12,13) These findings substantiate the occurrence of viremia in influenza virus infection and the extrapulmonary dissemination of the virus. Furthermore, influenza virus has been isolated from fetal heart, 10) placenta 14) and amniotic fluid, 10,15) suggesting the occurrence of fetal, placental and amniotic fluid infections with this virus.The influenza virus has not so far been isolated from the fetal membranes of a mother who died of influenza virus infection to our knowledge. However, the virus has been isolated from the placenta and amniotic fluid. 14,15) Additionally, organ cultures of human placenta allow influenza virus replication.16) The influenza virus may spread from the placenta to fetal membranes because they are continuous with the placenta. Furthermore, since amniotic fluid fills amniotic cavity formed by fetal membranes, influenza virus in amniotic fluid may influence the membrane cells. However, little is understood about the influence of influenza virus on human fetal membrane cells.We investigated the relationship between the influenza virus and cultured human fetal membrane cells by studying virus production and the apoptotic death of these cells in vitro. We found that the influenza virus induced cytocidal infection in chorion cells, persistent infection accompanied by virus production and cell survival in amnion cells, and that influenza virus-infected chorion cells became degraded through the apoptotic pathway. The present study demonstrates that the host cells make the choice to commit to degradation through this pathway. MATERIALS AND METHODSChemicals Ribavirin (1-b-D-ribofuranosyl-1,2,4-triazole-3-carboxamide) was purchased from Sigma (MO, U.S.A.).Influenza Virus Propagation and Plaque-Forming Assay Influenza virus type A (PR/8/34, H1N1) was propagated in the allantoic cavity of 11-d-old embryonal chicken eggs for 48 to 72 h at 35°C as described.2) Plaque-forming capacity was assayed on confluent monolayers of MDCK (NBL-2) cells (Human Science Research Resources Bank, Japan) as described.4) The chorioallantoic fluid contained 8ϫ10 8 plaque-forming units (PFU)/ml and the hemagglutination titer using human erythrocytes was 1 : 256. Cell Cultures and Virus Infection Human fetal membranes were prepared aseptically from placentas obtained by cesarean section in the month of normal parturition. Primary cult...

show abstract

Section: Resultsmentioning

confidence: 99%

Apoptosis in Cultured Human Fetal Membrane Cells Infected with Influenza Virus.

Uchide

Ohyama

Bessho³

et al. 2002

Biological & Pharmaceutical Bulletin

View full text Add to dashboard Cite

show abstract

“…Ribavirin has also been shown to have an inhibitory effect on viral polymerases (9,10). It has been suggested that ribavirin triphosphate could bind to the nucleotide-binding site of polymerases, thereby competitively inhibiting the binding of the nucleotides (11).…”

mentioning

confidence: 99%

Initial Binding of the Broad Spectrum Antiviral Nucleoside Ribavirin to the Hepatitis C Virus RNA Polymerase

Bougie

Bisaillon

2003

Journal of Biological Chemistry

View full text Add to dashboard Cite

Ribavirin is a broad spectrum antiviral nucleoside that displays activity against a variety of RNA and DNA viruses. Ribavirin is currently used in combination with interferon-␣ for the treatment of hepatitis C virus (HCV) infection and was recently shown to be directly incorporated by the HCV RNA polymerase into RNA products. This capacity ultimately leads to increased mutation rates and drastically reduces the viral fitness. As a first step toward elucidating the nature of the specific interaction between ribavirin and the HCV polymerase, we have utilized fluorescence spectroscopy to monitor precisely the binding of ribavirin triphosphate (RTP) to the viral polymerase. This spectroscopic approach allowed us to clearly separate the RTP binding activity from the concomitant catalytic steps. We report here the first detailed study of the binding kinetics and thermodynamic parameters involved in the interaction between RTP and an RNA polymerase. We demonstrate that RTP binds to the same active site as nucleotides. Furthermore, we provide evidence that the HCV polymerase cannot only bind to RTP but also to nonphosphorylated ribavirin, albeit with less affinity. By using various combinations of template-primers, we also demonstrate that base pairing is not involved in the initial binding of RTP to the HCV polymerase. Based on the results of circular dichroism and denaturation studies, we show that the RNA polymerase undergoes subtle conformational changes upon the binding of RTP, although the interaction does not significantly modify the stability of the protein. Finally, although metal ions are required for catalytic activity, they are not required for the initial binding of RTP to the polymerase. Such quantitative analyses are of primary importance for the rational design of new ribavirin analogues of potential therapeutic value and provide crucial insights on the interaction between RTP and the HCV RNA polymerase.Ribavirin is a broad spectrum antiviral nucleoside that displays activity against a variety of RNA and DNA viruses (1, 2). Once inside the cells, ribavirin is phosphorylated by cellular kinases, with ribavirin triphosphate as the major intracellular metabolite (3, 4). A number of possible mechanisms have been proposed over the years to account for the antiviral activity of ribavirin. Ribavirin has been shown to inhibit the host cell inosine monophosphate dehydrogenase, an enzyme involved in the de novo synthesis of GTP (5, 6). Because GTP is required for the transcription of viral genomes, and replication of RNA viruses, it has been assumed that the decrease in cytosolic concentration of GTP could affect the multiplication of viruses. Ribavirin has also been shown to modulate the host immune system by engendering a bias toward helper T-cells type 1 cytokine response (7,8). This would ultimately lead to an enhanced immune response against viral infections.Ribavirin has also been shown to have an inhibitory effect on viral polymerases (9, 10). It has been suggested that ribavirin triphosphate could bind to ...

show abstract

“…In fact, it has been shown that ribavirin inhibits influenza virus infection ex vivo and in vivo by inhibition of the viral polymerase (15,50). Through in vitro assays, the influenza A virus infection is inhibited by RTP but is not inhibited by RMP or by nonphosphorylated ribavirin (15).…”

Section: Discussionmentioning

confidence: 99%

“…Ribavirin has proven to be an inhibitor of several ortho-and paramyxoviruses (42). Furthermore, ribavirin inhibits in vitro and in vivo replication of influenza virus (15,33,49,50), the most typical member of the Orthomyxoviridae family.…”

mentioning

confidence: 99%

“…Ribavirin can be incorporated into nascent genomic RNA, causing an early termination of RNA synthesis or an error catastrophe through the misincorporation of cytidine and uridine instead of guanidine or adenine, which results in lethal mutagenesis (11,47). Ribavirin also inhibits HIV (35,40), vesicular stomatitis virus (VSV) (16), and La Crosse virus (7) polymerase; reovirus transcriptase (36); vaccinia virus guanylyltransferase (19); and influenza virus polymerase (15,50).…”

mentioning

confidence: 99%

See 1 more Smart Citation

Inhibitory Effect of a Nucleotide Analog on Infectious Salmon Anemia Virus Infection

Rivas‐Aravena

Vallejos‐Vidal²,

Martı́n

et al. 2011

J Virol

View full text Add to dashboard Cite

The infectious salmon anemia virus (ISAV), which belongs to the Orthomyxoviridae family, has been responsible for major losses in the salmon industry, with mortalities close to 100% in areas where Atlantic salmon (Salmo salar) is grown. This work studied the effect of ribavirin (1-␤-D-ribofuranosyl-1,2,3-triazole-3-carbaxaide), a broad-spectrum antiviral compound with proven ability to inhibit the replicative cycle of the DNA and RNA viruses. The results show that ribavirin was able to inhibit the infectivity of ISAV in in vitro assays. In these assays, a significant inhibition of the replicative viral cycle was observed with a 50% inhibitory concentration (IC 50 ) of 0.02 g/ml and an IC 90 of 0.4 g/ml of ribavirin. After ribavirin treatment, viral proteins were not detectable and a reduction of viral mRNA association with ribosomes was observed. Ribavirin does not affect the levels of EF1a, nor its association with polysomes, suggesting that the inhibition of RNA synthesis occurs specifically for the virus mRNAs and not for cellular mRNAs. Moreover, ribavirin caused a significant reduction in genomic and viral RNA messenger levels. The study of the inhibitory mechanism showed that it was not reversed by the addition of guanosine. Furthermore, in vivo assays showed a reduction in the mortality of Salmo salar by more than 90% in fish infected with ISAV and treated with ribavirin without adverse effects. In fact, these results show that ribavirin is an antiviral that could be used to prevent ISAV replication either in vitro or in vivo.

show abstract

Inhibition of Influenza Virus Ribonucleic Acid Polymerase by Ribavirin Triphosphate

Cited by 251 publications

References 20 publications

Apoptosis in Cultured Human Fetal Membrane Cells Infected with Influenza Virus.

Apoptosis in Cultured Human Fetal Membrane Cells Infected with Influenza Virus.

Initial Binding of the Broad Spectrum Antiviral Nucleoside Ribavirin to the Hepatitis C Virus RNA Polymerase

Inhibitory Effect of a Nucleotide Analog on Infectious Salmon Anemia Virus Infection

Contact Info

Product

Resources

About