Integrated Research Facility, Division of Clinical Research, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Frederick, Maryland, United States of America.
Emerging Viral Pathogens Section, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Frederick, Maryland, United States of America.
PLoS One. 2018 Mar 22;13(3):e0194880. doi: 10.1371/journal.pone.0194880. eCollection 2018.
Identifying effective antivirals for treating Ebola virus disease (EVD) and minimizing transmission of such disease is critical. A variety of cell-based assays have been developed for evaluating compounds for activity against Ebola virus. However, very few reports discuss the variable assay conditions that can affect the results obtained from these drug screens. Here, we describe variable conditions tested during the development of our cell-based drug screen assays designed to identify compounds with anti-Ebola virus activity using established cell lines and human primary cells. The effect of multiple assay readouts and variable assay conditions, including virus input, time of infection, and the cell passage number, were compared, and the impact on the effective concentration for 50% and/ or 90% inhibition (EC50, EC90) was evaluated using the FDA-approved compound, toremifene citrate. In these studies, we show that altering cell-based assay conditions can have an impact on apparent drug potency as measured by the EC50. These results further support the importance of developing standard operating procedures for generating reliable and reproducible in vitro data sets for potential antivirals.
确定有效的抗病毒药物来治疗埃博拉病毒病（EVD）并最大程度减少此类疾病的传播至关重要。已经开发了多种基于细胞的测定法来评估针对埃博拉病毒的化合物的活性。然而，很少有报道讨论可能影响从这些药物筛选中获得的结果的可变测定条件。在这里，我们描述了在开发用于鉴定具有抗埃博拉病毒活性的化合物的基于细胞的药物筛选测定法过程中测试的可变条件，这些测定法使用已建立的细胞系和人原代细胞。比较了多种测定读出值和可变测定条件（包括病毒输入，感染时间和细胞传代数）的影响，并使用经美国食品和药物管理局批准的化合物枸橼酸托瑞米芬评估了对 50%和/或 90%抑制有效浓度（EC50，EC90）的影响。在这些研究中，我们表明，改变基于细胞的测定条件会对通过 EC50 测量的药物效力产生影响。这些结果进一步支持为潜在的抗病毒药物制定生成可靠和可重复的体外数据集的标准操作程序的重要性。
