ChloroquineV1, Main, Exploration, bibRecord, 000D36

A Safe and Sensitive Antiviral Screening Platform Based on Recombinant Human Coronavirus OC43 Expressing the Luciferase Reporter Gene.

Identifieur interne : 000D36 ( Main/Exploration ); précédent : 000D35; suivant : 000D37

A Safe and Sensitive Antiviral Screening Platform Based on Recombinant Human Coronavirus OC43 Expressing the Luciferase Reporter Gene.

Auteurs : Liang Shen [République populaire de Chine] ; Yang Yang [République populaire de Chine] ; Fei Ye [République populaire de Chine] ; Gaoshan Liu [République populaire de Chine] ; Marc Desforges [Canada] ; Pierre J. Talbot [Canada] ; Wenjie Tan [République populaire de Chine]

Source :

Antimicrobial Agents and Chemotherapy [ 0066-4804 ] ; 2016-07-01.

RBID : Hal:pasteur-01351547

Abstract

Human coronaviruses (HCoVs) cause 15-30% of mild upper respiratory tract infections. However, no specific antiviral drugs are available to prevent or treat HCoV infections to date. Here, we developed four infectious recombinant HCoVs-OC43 (rHCoVs-OC43), which express the Renilla luciferase (Rluc) reporter gene. Among these four rHCoVs-OC43, rOC43-ns2DelRluc (generated by replacing ns2 with the Rluc gene) showed robust luciferase activity with only a slight impact on its growth characteristics. Additionally, this recombinant virus remained stable for at least 10 passages in BHK-21 cells. The rOC43-ns2DelRluc was comparable with its parental wild-type virus (HCoV-OC43-WT) with respect to the quantity of the antiviral activity of chloroquine and ribavirin. We showed that chloroquine strongly inhibited HCoV-OC43 replication in vitro, with an IC50 of 0.33 μM. However, ribavirin showed inhibition on HCoV-OC43 replication only at high concentrations which may not be applicable to humans in clinical treatment, with an IC50 of 10 μM. Furthermore, using a luciferase-based small interfering RNA (siRNA) screening assay, we identified double-stranded RNA-activated protein kinase (PKR) and DEAD-box RNA helicases (DDX3X) that exhibited antiviral activities, which were further verified by the use of HCoV-OC43-WT. Therefore, rOC43-ns2DelRluc represents a promising safe and sensitive platform for high throughput antiviral screening and quantitative analysis of viral replication.

Url:

https://hal-riip.archives-ouvertes.fr/pasteur-01351547

DOI: 10.1128/aac.00814-16

Affiliations:

Canada, République populaire de Chine

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to stream Hal, to step Checkpoint: 000071
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to stream Main, to step Curation: 000D36

Le document en format XML

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<front><div type="abstract" xml:lang="en">              <p>Human coronaviruses (HCoVs) cause 15-30% of mild upper respiratory tract infections. However, no specific antiviral drugs are available to prevent or treat HCoV infections to date. Here, we developed four infectious recombinant HCoVs-OC43 (rHCoVs-OC43), which express the Renilla luciferase (Rluc) reporter gene. Among these four rHCoVs-OC43, rOC43-ns2DelRluc (generated by replacing ns2 with the Rluc gene) showed robust luciferase activity with only a slight impact on its growth characteristics. Additionally, this recombinant virus remained stable for at least 10 passages in BHK-21 cells. The rOC43-ns2DelRluc was comparable with its parental wild-type virus (HCoV-OC43-WT) with respect to the quantity of the antiviral activity of chloroquine and ribavirin. We showed that chloroquine strongly inhibited HCoV-OC43 replication in vitro, with an IC50 of 0.33 μM. However, ribavirin showed inhibition on HCoV-OC43 replication only at high concentrations which may not be applicable to humans in clinical treatment, with an IC50 of 10 μM. Furthermore, using a luciferase-based small interfering RNA (siRNA) screening assay, we identified double-stranded RNA-activated protein kinase (PKR) and DEAD-box RNA helicases (DDX3X) that exhibited antiviral activities, which were further verified by the use of HCoV-OC43-WT. Therefore, rOC43-ns2DelRluc represents a promising safe and sensitive platform for high throughput antiviral screening and quantitative analysis of viral replication.</p>
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A Safe and Sensitive Antiviral Screening Platform Based on Recombinant Human Coronavirus OC43 Expressing the Luciferase Reporter Gene.

A Safe and Sensitive Antiviral Screening Platform Based on Recombinant Human Coronavirus OC43 Expressing the Luciferase Reporter Gene.

Source :

Abstract

Links toward previous steps (curation, corpus...)

Le document en format XML

Pour manipuler ce document sous Unix (Dilib)

Pour mettre un lien sur cette page dans le réseau Wicri