Ansarah-Sobrinho Camilo, Nelson Steevenson, Jost Christiane A, Whitehead Stephen S, Pierson Theodore C
Viral Pathogenesis Section, Laboratory of Viral Diseases, National Institutes of Health, Bethesda, MD, USA.
Virology. 2008 Nov 10;381(1):67-74. doi: 10.1016/j.virol.2008.08.021. Epub 2008 Sep 17.
Dengue virus (DENV) is a mosquito-borne flavivirus responsible for 50 to 100 million human infections each year, highlighting the need for a safe and effective vaccine. In this study, we describe the production of pseudoinfectious DENV reporter virus particles (RVPs) using two different genetic complementation approaches, including the creation of cell lines that release reporter viruses in an inducible fashion. In contrast to studies with West Nile virus (WNV), production of infectious DENV RVPs was temperature-dependent; the yield of infectious DENV RVPs at 37 degrees C is significantly reduced in comparison to experiments conducted at lower temperatures or with WNV. This reflects both a significant reduction in the rate of infectious DENV RVP release over time, and the more rapid decay of infectious DENV RVPs at 37 degrees C. Optimized production approaches allow the production of DENV RVPs with titers suitable for the study of DENV entry, assembly, and the analysis of the humoral immune response of infected and vaccinated individuals.
登革病毒（DENV）是一种由蚊子传播的黄病毒，每年导致5000万至1亿人感染，凸显了对安全有效疫苗的需求。在本研究中，我们描述了使用两种不同的基因互补方法生产假感染性登革病毒报告病毒颗粒（RVP），包括创建以可诱导方式释放报告病毒的细胞系。与西尼罗河病毒（WNV）的研究不同，感染性登革病毒RVP的生产是温度依赖性的；与在较低温度下进行的实验或WNV实验相比，37摄氏度时感染性登革病毒RVP的产量显著降低。这既反映了随着时间推移感染性登革病毒RVP释放速率的显著降低，也反映了37摄氏度时感染性登革病毒RVP更快的衰减。优化的生产方法能够生产出滴度适合研究登革病毒进入、组装以及分析感染和接种个体体液免疫反应的登革病毒RVP。
