Arbuckle Jesse H, Turner Anne-Marie W, Kristie Thomas M
Laboratory of Viral Diseases, National Institutes of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland.
Curr Protoc Microbiol. 2014 Nov 3;35:14E.6.1-21. doi: 10.1002/9780471729259.mc14e06s35.
As with all Herpesviruses, Herpes simplex virus (HSV) has both a lytic replication phase and a latency-reactivation cycle. During lytic replication, there is an ordered cascade of viral gene expression that leads to the synthesis of infectious viral progeny. In contrast, latency is characterized by the lack of significant lytic gene expression and the absence of infectious virus. Reactivation from latency is characterized by the re-entry of the virus into the lytic replication cycle and the production of recurrent disease. This unit describes the establishment of the mouse sensory neuron model of HSV-1 latency-reactivation as a useful in vivo system for the analysis of mechanisms involved in latency and reactivation. Assays including the determination of viral yields, immunohistochemical/immunofluorescent detection of viral antigens, and mRNA quantitation are used in experiments designed to investigate the network of cellular and viral proteins regulating HSV-1 lytic infection, latency, and reactivation.
与所有疱疹病毒一样,单纯疱疹病毒(HSV)具有裂解复制阶段和潜伏-激活周期。在裂解复制期间,存在有序的病毒基因表达级联反应,导致传染性病毒后代的合成。相比之下,潜伏的特征是缺乏明显的裂解基因表达且不存在传染性病毒。潜伏激活的特征是病毒重新进入裂解复制周期并产生复发性疾病。本单元描述了HSV-1潜伏-激活小鼠感觉神经元模型的建立,该模型是用于分析潜伏和激活相关机制的有用体内系统。在旨在研究调节HSV-1裂解感染、潜伏和激活的细胞和病毒蛋白网络的实验中,使用了包括病毒产量测定、病毒抗原的免疫组织化学/免疫荧光检测以及mRNA定量在内的检测方法。