Characterization of a novel rabbit herpes simplexvirus: a possible model for unde

描述（由申请方提供）：发现一种疱疹病毒（称为LHV-4）与一种疾病相关，该疾病影响阿拉斯加州安克雷奇附近一个农场的55只兔子中的一半以上。大约28%的受影响动物死亡或因疾病严重而被安乐死。感染兔皮肤提取物在培养的兔皮肤和肾细胞以及Vero细胞中产生合胞体和细胞溶解。使用针对保守疱疹病毒基因的变性引物的PCR用于从纯化的病毒DNA扩增序列。这些产物的测序允许设计特异性引物，从而可以确定许多基因的完整序列数据。对该序列数据的分析表明，该病毒是疱疹病毒单属的成员，与牛疱疹病毒2型、HSV-1、HSV-2和猕猴疱疹病毒最密切相关。新西兰白色家兔实验性暴露于分离株导致严重眼病以及脾和淋巴结坏死。此外，在急性感染期间（感染后5天内），通过PCR在多种组织中鉴定出病毒DNA，但仅在感染后14天后在三叉神经节（TG）中可持续检测到。Swiss韦伯斯特小鼠以1 × 103 PFU/眼的剂量实验性暴露于分离株导致轻度眼病和发热。急性感染时脾组织中可检测到病毒DNA，感染后20天，TG组织中可检测到病毒DNA。α疱疹病毒的独特特征之一是它们在TG中潜伏，除了潜伏相关转录基因（LAT）外没有基因表达。LAT在潜伏期期间的作用仍然没有很好地理解。LAT基因可能是理解HSV-1或HSV-2潜伏期再激活机制的关键。本提案中概述的研究目的是研究这种新型疱疹病毒的LAT基因。为了实现这一点，我们将测序和注释LHV-4的基因组，并表征LAT区域。由于其与灵长类病毒谱系的密切关系，以及其感染兔子的能力，关于该病毒潜伏期的信息可能为理解人类疱疹病毒感染的潜伏期提供自然宿主模型系统。公共卫生相关性：在这项研究应用中，我们建议对最近发现的一种与HSV-1和2密切相关的兔单纯疱疹病毒的基因组进行测序。本研究的目的是表征潜伏相关转录基因（LAT）区域，并研究LAT在自然宿主模型系统中的潜伏期中的作用。.

DESCRIPTION (provided by applicant): A herpesvirus, referred to as LHV-4, was found to be associated with a disease affecting over half of the 55 rabbits on a farm near Anchorage, Alaska. About 28% of the affected animals died or were euthanized because of the severity of the illness. Extracts from the skin of infected rabbits produced syncytia and cell lysis in cultured rabbit skin and kidney cells, and in Vero cells. PCR using degenerative primers to conserved herpesvirus genes was used to amplify sequences from purified viral DNA. Sequencing of these products allowed the design of specific primers so that complete sequence data for a number of genes could be determined. Analysis of this sequence data indicates that the virus is a member of the simplex genus of herpesviruses and is most closely related to bovine herpesvirus 2, HSV-1, HSV-2, and cercopithecine herpesvirus. Experimental exposure of New Zealand White rabbits to the isolate resulted in severe eye disease and necrosis of the spleen and lymph nodes. In addition, viral DNA was identified in a variety of tissues by PCR during the acute infection (within 5 days post- infection), but was only detectable consistently in trigeminal ganglia (TG) after 14 days post- infection. Experimental exposure of Swiss Webster mice to the isolate at 1X103 PFU/eye resulted in mild eye disease and fever. Viral DNA was detected in spleen tissue during the acute infection, and was detectable in the TG after 20 day post-infection. One of the unique features of alphaherpes virus is that they become latent in the TG with no gene expression, except for the latency associated transcript gene (LAT). The role of LAT during latency is still not well understood. The LAT gene may be the key to understand the mechanism of HSV-1 or HSV-2 latency reactivation. The purpose of the research outlined in this proposal is to investigate LAT gene of this novel herpesvirus. To accomplish this we will sequence and annotate the genomes of LHV-4 and characterize the LAT region. Because of its close relationship to the primate virus lineage, and its ability to infect rabbits, information on latency in this virus may provide a natural host model system for understanding latency in herpesvirus infections of humans. PUBLIC HEALTH RELEVANCE: In this research application we propose to sequence the genome of a recently discovered Simplex herpesvirus pathogenic for rabbits that is closely related to HSV-1 and 2. The goal of this research is to characterize the latency associated transcript gene (LAT) region and investigate the role of LAT during latency in a natural host model system. .