THE ROLE OF CYTOMEGALOVIRUS PHOSPHOPROTEIN 65 IN VIRUS REPLICATON IN VIVO

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 巨细胞病毒(CMV)pp65蛋白调节宿主免疫反应，同时也是细胞免疫的主要抗病毒靶点。Pp65是恒河猴CMV(RhCMV)和人CMV(HCMV)被膜中含量最丰富的成分，是衣壳和被膜之间的富含蛋白质的一层，释放到细胞质中。有趣的是，pp65对于这两种病毒的体外生长都不是必需的。这使得我们能够产生pp65缺陷的RhCMV，以检测1)pp65‘S的免疫调节功能是否对病毒在体内的存活至关重要；或2)pp65的免疫原性是否限制了病毒在健康动物中的复制。用突变的RhCMV感染恒河猴(RMS)，可以检测pp65在体内CMV感染中的重要性。我们将用pp65缺失的RhCMV感染CMV-NA猕猴，并评估该病毒是否能够建立原发和慢性感染，如果是的话，病毒学和免疫学参数与之前在感染野生型RhCMV期间所观察到的比较。根据结果，我们将评估pp65缺失的病毒是否能够再次感染这些动物。如果生产性感染需要pp65编码的功能，我们预计突变的复制要么无法检测到，要么会减少。相反，如果pp65的免疫原性对宿主免疫反应控制病毒感染的能力至关重要，我们预测pp65缺失的病毒的复制将比野生型更大。这一结果对CMV疫苗和抗病毒药物的设计以及CMV作为疫苗载体的发展都有意义。

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. Cytomegalovirus (CMV) protein pp65 modulates the host immune response yet simultaneously is the primary antiviral target of cellular immunity. pp65 is the most abundant component of the Rhesus CMV (RhCMV) and human CMV (HCMV) tegument, a protein-rich layer between the capsid and the envelope that is released into the cytoplasm. Interestingly, pp65 is not required for in vitro growth of either virus. This allowed us to generate a pp65-deficient RhCMV to examine whether 1) pp65's immunomodulatory functions are critical for virus survival in vivo; or 2) pp65 immunogenicity limits virus replication in healthy animals. Infection of rhesus macaques (RMs) with mutant RhCMV will allow examination of the importance of pp65 for CMV infection in vivo. We will infect CMV-na¿ve macaques with pp65-deleted RhCMV and evaluate whether this virus is able to establish primary and chronic infection and, if so, how virological and immunological parameters compare to previous observations made during infection with wild type RhCMV. Depending on the outcome, we will evaluate whether pp65-deleted virus is able to re-infect these animals. If pp65-encoded functions are required for productive infection, we expect replication of the mutant to be either undetectable or diminished. Conversely, if pp65 immunogenicity is crucial to the ability of the host immune response to control virus infection we predict that replication of the pp65-deleted virus will be greater than that for wild type. The outcome has implications both for the design of CMV vaccines and antivirals as well as the development of CMV as a vaccine vector.