AIM:To evaluate the antiviral potency of a new antihepatitis C virus(HCV)antiviral agent targeting the cellular autophagy machinery.METHODS:Non-infected liver slices,obtained from human liver resection and cut in 350...AIM:To evaluate the antiviral potency of a new antihepatitis C virus(HCV)antiviral agent targeting the cellular autophagy machinery.METHODS:Non-infected liver slices,obtained from human liver resection and cut in 350μm-thick slices(2.7×106 cells per slice)were infected with cell culture-grown HCV Con1b/C3 supernatant(multiplicity of infection=0.1)cultivated for up to ten days.HCV infected slices were treated at day 4 post-infection with GNS-396 for 6 d at different concentrations.HCV replication was evaluated by strand-specific real-time quantitative reverse transcription-polymerase chain reaction.The infectivity titers of supernatants were evaluated by foci formation upon inoculation into naive Huh-7.5.1 cells.The cytotoxic effect of the drugs was evaluated by lactate dehydrogenase leakage assays.RESULTS:The antiviral efficacy of a new antiviral drug,GNS-396,an autophagy inhibitor,on HCV infection of adult human liver slices was evidenced in a dosedependent manner.At day 6 post-treatment,GNS-396 EC50 was 158 nmol/L without cytotoxic effect(compared to hydroxychloroquine EC50=1.17μmol/L).CONCLUSION:Our results demonstrated that our ex vivo model is efficient for evaluation the potency of autophagy inhibitors,in particular a new quinoline derivative GNS-396 as antiviral could inhibit HCV infection in a dosedependent manner without cytotoxic effect.展开更多
基金Supported by The Institut National de la Sante et de la Recherche Medicale(INSERM,France)the personal support of Professor Jean-Francois Delfraissy as Director of the French Agency,Agence Nationale de Recherches sur le Sida et les hepatites virales(ANRS)
文摘AIM:To evaluate the antiviral potency of a new antihepatitis C virus(HCV)antiviral agent targeting the cellular autophagy machinery.METHODS:Non-infected liver slices,obtained from human liver resection and cut in 350μm-thick slices(2.7×106 cells per slice)were infected with cell culture-grown HCV Con1b/C3 supernatant(multiplicity of infection=0.1)cultivated for up to ten days.HCV infected slices were treated at day 4 post-infection with GNS-396 for 6 d at different concentrations.HCV replication was evaluated by strand-specific real-time quantitative reverse transcription-polymerase chain reaction.The infectivity titers of supernatants were evaluated by foci formation upon inoculation into naive Huh-7.5.1 cells.The cytotoxic effect of the drugs was evaluated by lactate dehydrogenase leakage assays.RESULTS:The antiviral efficacy of a new antiviral drug,GNS-396,an autophagy inhibitor,on HCV infection of adult human liver slices was evidenced in a dosedependent manner.At day 6 post-treatment,GNS-396 EC50 was 158 nmol/L without cytotoxic effect(compared to hydroxychloroquine EC50=1.17μmol/L).CONCLUSION:Our results demonstrated that our ex vivo model is efficient for evaluation the potency of autophagy inhibitors,in particular a new quinoline derivative GNS-396 as antiviral could inhibit HCV infection in a dosedependent manner without cytotoxic effect.
