THE IMPACT OF IRF-3-DEPENDENT MECHANISMS ON THE REPLICATION AND VIRULENCE OF HSV

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目及 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者的研究机构。 干扰素调节因子3（IRF 3）在IFN应答的诱导阶段至关重要。然而，它在控制某些病毒（包括HSV-1）复制中的作用是神秘的。 许多HSV基因在体外干扰IRF-3的定位和功能，表明IRF-3对控制HSV复制至关重要。 因此，IRF-3的缺陷将被预测为排除早期识别途径的功能，从而影响HSV-1的复制。奇怪的是，IRF-3缺陷型小鼠（IRF 3-/-）对HSV-1的易感性没有增加，并且在原发性IRF-3-/-MEF中没有观察到HSV复制的变化。与此一致，我们已经表明，主要MEF不施加这样的控制。然而，其他最近的初步数据显示HSV在IRF 3-/-树突细胞中的生长显著增加，表明来自免疫系统的细胞对HSV复制发挥IRF-3依赖性控制。 我们推断有两种可能的假设来解释这些观察结果。 首先，HSV完全控制IRF-3活性，以至于在某些细胞中，IRF-3变得多余。第二，存在通过IRF-3依赖性机制控制HSV体外和体内复制以及体内毒力的组织特异性。 因此，本提案的首要目标是测试这些假设并区分它们。 具体目标1：测量和评估IRF-3依赖性机制对体外免疫系统原代细胞中HSV复制的影响。具体目标2：在HSV发病机制和体内潜伏期小鼠模型中，测量和评估IRF-3依赖性机制对HSV复制和毒力的影响。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Interferon regulatory factor 3 (IRF3) is critical in the induction phase of the IFN response. Its role, however, in the control of replication of certain viruses, including HSV-1, is enigmatic. A number of HSV genes interfere with the localization and function of IRF-3 in vitro, suggesting that IRF-3 is critical for controlling HSV replication. The deficiency ofIRF-3 would therefore be predicted to preclude the function of early recognition pathways and thereby impact HSV-1 replication. Paradoxically,IRF-3 deficient mice (IRF3-/-) show no increased susceptibility to HSV-1,and no changes in HSV replication in primary IRF-3-/- MEFs have been observed. In agreement with this, we have shown that primary MEFs do not exert such control. Other recent preliminary data, however, showed significantly increased growth of HSV in IRF3-/- dendritic cells, suggesting that cells derived from the immune system exert IRF-3 dependent control over HSV replication. We reasoned that there were two possible hypotheses to explain these observations. First, that HSV controls IRF-3 activity so completely that, in certain cells, IRF-3 is rendered redundant. Second, that there is a tissue-specificity to the control of HSV replication in vitro and in vivo, and virulence in vivo by IRF-3- dependent mechanisms. The over-arching goal of this proposal is therefore, to test these hypotheses and distinguish between them. Specific aim 1: To measure and assess the impact of IRF-3- dependent mechanisms upon the replication of HSV in primary cells derived from the immune system in vitro. Specific aim 2: To measure and assess the impact of IRF-3- dependent mechanisms upon the replication and virulence of HSV in mouse models of HSV pathogenesis and latency in vivo.