Viral Genes Essential for MHV-68 Persistent Infection in the Spleen

γ疱疹病毒亚家族的成员在淋巴细胞中建立潜伏感染并在被感染的宿主中引起良性或恶性肿瘤的能力是不同的。在这个亚家族中有两种人类病毒，Epstein-Barr病毒（EBV）和人类疱疹病毒-8/卡波西肉瘤相关疱疹病毒（HHV-8/KSHV）。鼠γ疱疹病毒-68 （MHV-68）是同一亚家族的另一个成员。由于EBV和KSHV不能在细胞培养物或小型模型动物中有效复制，因此MHV-68可以作为研究病毒和宿主在病毒复制和发病机制中的作用的良好模型系统。在MHV-68系统中可以确定病毒基因的功能，这将为人类伽玛疱疹病毒的相关基因提供有用的信息。我们以发现为导向的方法的长期目标是通过整个病毒基因组的突变来确定每个病毒基因的功能。通过将标记标记的转座子随机插入到MHV-68细菌人工染色体中来诱变MHV-68基因组。细胞培养中病毒复制所必需的病毒基因已被确定。这个探索性R21应用程序的目的是鉴定在脾脏中建立长期持续性感染所必需的病毒基因。这项研究将是首次对病毒基因功能进行全面分析。这些结果将使我们能够通过假设驱动的研究来跟踪特定的病毒基因。这将为我们研究病毒持续存在的机制打开一扇新的窗口，为发现潜在的新治疗靶点提供了可能。此外，确定持续感染所必需的病毒基因将使我们能够构建具有复制能力但不能建立持续感染的重组病毒。这些病毒将强烈刺激免疫反应，但不会建立持续感染以引起疾病，因此是一种潜在的疫苗策略。

DESCRIPTION: Viral genes essential for MHV-68 Persistent Infection in the Spleen Members of the gammaherpesvirus subfamily are distinct in their ability to establish latent infections in the lymphocytes and cause benign or malignant tumors in infected hosts. There are two human viruses in this subfamily, Epstein-Barr virus (EBV) and human herpesvirus-8/Kaposi's sarcoma-associated herpesvirus (HHV-8/KSHV). Murine gammaherpesvirus-68 (MHV-68) is another member of the same subfamily. Since EBV and KSHV cannot effectively replicate in cell cultures or small model animals, MHV-68 serves as an excellent model system for studying the role of both the virus and the host in viral replication and pathogenesis. The function of viral genes can be determined in the MHV-68 system, which should provide instrumental information for the related genes in human gammaherpesviruses. The long-term goal of our discovery-oriented approach is to determine the function of each viral gene by mutagenesis of the whole viral genome. MHV-68 genome is mutagenized by random insertion of signature- tagged transposons into an MHV-68 bacterial artificial chromosome. The viral genes essential for viral replication in cell cultures have been identified. The objective of this exploratory R21 application is to identify viral genes essential for establishment of long-term persistent infection in the spleen. This study will be the first comprehensive analysis of viral gene function. The results will allow us to follow up on specific viral genes with hypothesis driven study. This will open a new window for us to look into the mechanism of viral persistence, which provides possibilities of identifying potential new therapeutic targets. Furthermore define the viral genes essential for persistent infection will allow us to construct recombinant viruses that are replication competent but cannot establish persistent infection. These viruses will strongly stimulate immune responses but will not establish persistent infection to cause diseases, thus a potential vaccine strategy.