Grosche Linda, Döhner Katinka, Düthorn Alexandra, Hickford-Martinez Ana, Steinkasserer Alexander, Sodeik Beate
Department of Immune Modulation, Universitätsklinikum Erlangen, Erlangen, Germany.
Institute of Virology, OE5230, Hannover Medical School, Hannover, Germany.
Bio Protoc. 2019 Dec 5;9(23):e3441. doi: 10.21769/BioProtoc.3441.
Given the endemic seroprevalence of herpes simplex viruses (HSV), its associated human diseases, and the emergence of acyclovir-resistant strains, there is a continuous need for better antiviral therapies. Towards this aim, identifying mechanistic details of how HSV-1 manipulates infected cells, how it modulates the immune responses, and how it causes diseases are essential. Measuring titers and growth kinetics of clinical isolates and viral mutants are important for a thorough characterization of viral phenotypes and . We provide protocols for the preparation as well as titration of HSV-1 stocks, and explain how to perform single-step growth curves to characterize the functions of viral proteins or host factors during infection. In particular, we describe methods to prepare and characterize high-titer HSV-1 stocks with low genome to titer ratios that are required for infection studies in cell culture and animal experiments.
鉴于单纯疱疹病毒（HSV）的地方性血清流行率、其相关的人类疾病以及阿昔洛韦耐药菌株的出现，一直需要更好的抗病毒疗法。为了实现这一目标，确定HSV-1如何操纵受感染细胞、如何调节免疫反应以及如何引发疾病的机制细节至关重要。测量临床分离株和病毒突变体的滴度和生长动力学对于全面表征病毒表型很重要。我们提供了HSV-1毒株的制备和滴定方案，并解释了如何进行单步生长曲线以表征感染期间病毒蛋白或宿主因子的功能。特别是，我们描述了制备和表征低基因组与滴度比的高滴度HSV-1毒株的方法，这是细胞培养和动物实验感染研究所需的。
