Zhu Qin-Chang, Wang Yi, Peng Tao
State Key Laboratory for Respiratory Disease, Laboratory of Viral Immunology, Guangzhou Institute of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou Science Park, Guangzhou, China.
J Biomol Screen. 2010 Sep;15(8):1016-20. doi: 10.1177/1087057110372804. Epub 2010 Jun 14.
Most of the current antiherpetics target viral DNA polymerase, but with the emergence of drug-resistant viruses, antiherpetics with different targets have become necessary. Inhibition of herpes simplex virus (HSV) replication at the early stages of infection minimizes cytotoxicity and immune suppression induced by HSV infection. In this report, quantitative reporter systems that use recombinant HSV and a stably transfected cell line were developed for the screening of agents targeting the early stages of HSV infection. The reporter genes in both systems were directed by HSV immediate-early (IE) promoters, so considerably less time was required for the quantification of HSV infection than the traditional plaque reduction assay. The results show that both reporter assays were sensitive to antiherpetic screening. Both assays were quantitative, rapid, easy to perform, and highly adaptable for automatic high-throughput screening. Exploiting the flexibility of these 2 assays, modified assays were also proposed for the detailed analysis of antiherpetic mechanisms.
目前大多数抗疱疹病毒药物都靶向病毒DNA聚合酶,但随着耐药病毒的出现,具有不同靶点的抗疱疹病毒药物变得必不可少。在感染早期抑制单纯疱疹病毒(HSV)复制可将HSV感染诱导的细胞毒性和免疫抑制降至最低。在本报告中,开发了使用重组HSV和稳定转染细胞系的定量报告系统,用于筛选靶向HSV感染早期阶段的药物。两个系统中的报告基因均由HSV立即早期(IE)启动子指导,因此与传统的蚀斑减少试验相比,定量HSV感染所需的时间大大缩短。结果表明,两种报告基因检测方法对抗疱疹病毒药物筛选均敏感。两种检测方法均具有定量、快速、易于操作且高度适用于自动高通量筛选的特点。利用这两种检测方法的灵活性,还提出了改进的检测方法用于抗疱疹病毒药物作用机制的详细分析。