Rega Institute for Medical Research, Department of Microbiology and Immunology, Minderbroedersstraat 10, 3000 Leuven, Belgium.
Antiviral Res. 2013 May;98(2):325-31. doi: 10.1016/j.antiviral.2013.03.016. Epub 2013 Mar 23.
Three different antiviral assays were developed for the in vitro screening of inhibitors of the hepatitis A virus (HAV) of which (i) a cytopathic effect reduction assay suitable for medium-to-high-throughput screening and (ii) two virus yield reduction assays (based on quantification of viral RNA) for genotypes IB and IIIA. The assays were validated for antiviral studies with interferon-alpha (IFNα) and amantadine HCl, two known inhibitors of HAV replication. IFNα effectively inhibited HAV replication, whereas the activity of amantadine HCl appeared to be strain-dependent. Employing these assays, we assessed the effect of the known enterovirus inhibitors pleconaril, rupintrivir and enviroxime on HAV replication. Pleconaril exhibited some very moderate activity, the effect of rupintrivir proved to be strain-dependent. Enviroxime did not inhibit HAV replication, suggesting that phosphatidylinositol-4-kinase IIIβ is not crucial in the HAV life cycle.
开发了三种不同的抗病毒测定法用于体外筛选甲型肝炎病毒 (HAV) 的抑制剂，其中 (i) 适合中高通量筛选的细胞病变效应减少测定法，以及 (ii) 两种基于病毒 RNA 定量的基因型 IB 和 IIIA 的病毒产量减少测定法。该测定法已通过干扰素-α (IFNα) 和金刚烷胺盐酸盐（两种已知的 HAV 复制抑制剂）的抗病毒研究得到验证。IFNα 可有效抑制 HAV 复制，而金刚烷胺盐酸盐的活性似乎取决于毒株。利用这些测定法，我们评估了已知肠道病毒抑制剂普乐可复、鲁比那韦和恩替诺特对 HAV 复制的影响。普乐可复表现出一些非常温和的活性，鲁比那韦的效果证明取决于毒株。恩替诺特不抑制 HAV 复制，表明磷脂酰肌醇-4-激酶 IIIβ 并非 HAV 生命周期中的关键酶。
