MODULATION OF INNATE IMMUNE RESPONSES BY CYTOMEGALOVIRUS

这个子项目是许多利用资源的研究子项目之一 由NIH/NCRR资助的中心拨款提供。子项目的主要支持 而子项目的主要调查员可能是由其他来源提供的， 包括其它NIH来源。 列出的子项目总成本可能 代表子项目使用的中心基础设施的估计数量， 而不是由NCRR赠款提供给子项目或子项目工作人员的直接资金。 在一个有竞争力的补充父母补助金（AI 070890），我们研究是否RhCMV缺乏pp 65和pp 71是能够在恒河猴的原发性和再感染。产生pp 65缺失的RhCMV并检查其体外和体内复制。未观察到缺陷，这表明尽管pp 65是病毒体的主要结构组分，但其对于体外生长不是必需的。我们还表明，pp 65缺失的病毒可以保护免受缺乏T细胞逃避基因的病毒的攻击。这表明pp 65特异性T细胞不需要保护。我们还产生了pp 71缺失的RhCMV。由于已知pp 71启动立即早期基因表达的功能，该缺失突变体在体外高度减毒。有趣的是，尽管有这种主要的生长缺陷，pp 71缺失的病毒在感染恒河猴后诱导了针对CMV以及针对来自SIV的异源抗原的正常免疫应答。然而，在感染动物的分泌物中检测不到pp 71缺失的病毒。这一结果表明，pp 71缺失的HCMV可用作活复制疫苗和疫苗载体的安全替代物。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. In a competitive supplement to the parent grant (AI070890), we examine whether RhCMV lacking pp65 and pp71 is capable of primary and re-infection in rhesus macaques. A pp65-deleted RhCMV was generated and examined for replication in vitro and in vivo. No defect was observed suggesting that, despite being a major structural component of the virion, pp65 is not essential for growth in vitro. We also showed that the pp65-deleted virus can protect against challenge with a virus that lacks T cell evasion genes. This suggests that that pp65-specific T cells are not required for protection. We also generated a pp71-deleted RhCMV. This deletion mutant is highly attenuated in vitro due to the known function of pp71 to kick-start immediate early gene expression. Interestingly, despite this major growth defect, the pp71-deleted virus induced a normal immune response against CMV as well as against heterologous antigens from SIV upon infection of rhesus macaques. However, pp71-deleted virus was undetectable in the secretions of infected animals. This result suggests that a pp71-deleted HCMV could be used as a safe alternative for live replicating vaccines and vaccine vectors.