Genetic Analysis of Interactions Between Oral Innate Immunity and a Herpesvirus

描述（由申请人提供）：口腔是与外界接触的独特的身体部位，因此不断遇到各种各样的微生物。疱疹病毒是人类重要的常驻口腔病原体。然而，对于宿主在口腔中的初次感染、潜伏期和再激活过程中如何感知和响应疱疹病毒，尤其是先天免疫系统，知之甚少。病毒及其免疫活性宿主的共存需要病毒复制和宿主清除之间的微妙平衡。阐明先天免疫系统和疱疹病毒之间的相互作用将为了解如何实现和调节平衡提供见解。出于多种原因，鼠伽马疱疹病毒 68 (MHV-68) 感染的小鼠模型为探索口腔中病毒与宿主的相互作用提供了一个极好的系统。首先，口服给药后，MHV-68 在口鼻腔和唾液腺中复制。其次，可以使用合适的遗传系统来诱变 MHV-68 病毒基因组。第三，MHV-68感染多种实验室小鼠品系，这使得可以在体内研究病毒感染期间细胞基因的作用。 我们的发现驱动方法的长期目标是通过使用基因敲除小鼠和一组 MHV-68 突变病毒来确定病毒和细胞蛋白在病毒相关发病机制中的作用。 MHV-68 基因组因转座子的插入而被随机破坏。有 32 个病毒 ORF 不是在成纤维细胞中复制所必需的，并且可能参与调节病毒与宿主的相互作用。这一探索性 R21 应用的目的是使用口腔 MHV-68 感染的小鼠模型来定义 TLR 的体内传感和防御作用及其信号通路。此外，我们将系统地筛选能够抑制细胞中抗病毒反应激活的病毒基因，这是了解病毒与宿主相互作用的重要一步。

DESCRIPTION (provided by applicant): The oral cavity represents a unique body part that is in contact with the external world and therefore constantly encounters a vast diverse of microbes. Herpesviruses are significant resident oral pathogens in humans. However, little is known about how the host senses and responds to herpesviruses during primary infection, latency and reactivation in the oral cavity, especially in regards to the innate immune system. Coexistence of a virus and its immunocompetent host entails a delicate balance between viral replication and host clearance. Elucidating the interactions between the innate immune system and herpesviruses will provide insights to how the balance is accomplished and regulated. A mouse model of murine gammaherpesvirus-68 (MHV-68) infection provides an excellent system to explore virus-host interactions in the oral cavity for several reasons. First, following oral administration, MHV-68 replicates in the oronasal cavity and the salivary gland. Second, an amenable genetic system is available to mutagenize the MHV-68 viral genome. Third, MHV-68 infects a variety of laboratory strains of mice, which allows the role of cellular genes during viral infection to be studied in vivo. The long-term goal of our discovery driven approach is to determine the role of viral and cellular proteins in virus-associated pathogenesis by employing knock-out mice and a collection of MHV-68 mutant viruses. The MHV-68 genome is randomly disrupted by an insertion of a transponson. There are 32 viral ORFs that are not required for replication in fibroblast cells and potentially involved in modulating virus-host interactions. The object of this exploratory R21 application is to define the in vivo sensing and defending role of TLR and its signaling pathway using a mouse model of oral MHV-68 infection. Furthermore, we will systematically screen viral genes that are capable of inhibiting the activation of anti-viral responses in cells, an essential step for understanding virus-host interactions.