CMV VECTOR DESIGN AND DEVELOPMENT

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。 该项目是盖茨基金会和国际艾滋病疫苗倡议(IAVI)赞助的艾滋病疫苗开发大型联盟的一部分。该联盟的一个主要目标是利用恒河猴模型开发艾滋病疫苗的新方法，以评估有效性，特别是比较相同挑战方案中所有方法的有效性。本项目的具体目标是设计、构建和测试经过免疫原性和安全性优化的基因缺失的RhCMV载体，然后在为整个联盟制定的标准疗效方案中对野生型和基因缺失的RhCMV载体进行比较评估。我们将使用腺病毒和腺相关病毒(AAV)载体作为异源启动子或Boost，在异源启动子-Boost协议中单独测试RhCMV载体。因此，这个项目将有3个“活动”。在活动#1(“CMV载体设计和开发”)中，我们将1)构建和评估缺少一个或多个“非必需”宿主调节/免疫逃避基因的RhCMV突变体，2)设计和构建能够同时表达多个SIV基因的表达插入体(和载体)和允许在体内控制载体复制的“安全”插入体(例如，其生长依赖于或被诸如多西环素的高生物利用度药物终止的载体)，3)作为第一次或第二次疗效试验(见下文)的一部分，对使用这些载体的猴子中的RhCMV载体感染进行病毒学和病理学评估，以及4)开发和构建类似于最佳RhCMV设计的人巨细胞病毒(HCMV)的减毒/修饰突变体。在活动#2中，我们将评估“野生型”RhCMV载体的免疫原性、病理生物学和保护效果，无论是否有腺病毒(Ad)5增强。在活动#3中，我们将评估使用或不使用AAV启动的减毒(基因修饰或基因缺失)的RhCMV载体的免疫原性、病理生物学和保护效果。

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. This project is part of a large consortium on AIDS vaccine development sponsored by the Gates Foundation and the International AIDS Vaccine Initiative (IAVI). A major goal of the consortium is to develop new approaches to AIDS vaccine using the rhesus macaque model to assess efficacy and in particular compare efficacy of all approaches in an identical challenge protocol. The specific goal of this project is to design, construct and test gene-deleted RhCMV vectors that have been optimized for immunogenicity and safety, and then comparatively assess both wildtype RhCMV vectors and gene-deleted RhCMV vectors in the standard efficacy protocol developed for the entire consortium. RhCMV vectors will be tested alone as well as in heterologous prime-boost protocols using adenovirus and adeno-associated viral (AAV) vectors as the heterologous prime or boost. Thus this project will have 3 "Activities". In Activity #1 ("CMV Vector Design and Development"), we will 1) construct and evaluate RhCMV mutants that lack one or more "non-essential" host modulation/immune evasion genes, 2) design and construct expression inserts (and vectors) capable of simultaneous expression of multiple SIV genes and "safety" inserts that will allow in vivo control of vector replication with administration of an external drug (e.g., vectors whose growth is either dependent on or terminated by a drug of high bioavailability such as doxycycline), 3) provide virologic and pathologic assessment of RhCMV vector infections in the monkeys administered these vectors as part of the first or second efficacy trials (see below), and 4) develop and construct an attenuated/modified mutant of human cytomegalovirus (HCMV) analogous to the optimal RhCMV design. In Activity #2, we will assess the immunogenicity, pathobiology and protective efficacy of "wildtype" RhCMV vectors, with or without Adenovirus (Ad) 5 Boosting. In Activity #3, we will assess the immunogenicity, pathobiology and protective efficacy of attenuated (gene-modified or gene-deleted) RhCMV vectors, with or without AAV priming.