Generation and in vitro/vivo evaluation of an R5-tropic SHIV library

描述（由申请人提供）：HIV-1是人类艾滋病的主要原因，不能在大多数非人类物种中复制。因此，最实用的AIDS动物模型由用表达HIV-1包膜（SHIV）的SIVMAC或源自SIVMAC的嵌合体感染恒河猴组成。然而，这两种模式都有特定的局限性。在SHIV的情况下，迄今为止开发的大多数嵌合体使用CXCR 4共受体（X4-tropic），而大多数主要HIV-1分离株使用CCR 5共受体（R5-tropic）。寻找在动物中复制并引起疾病的R5嗜性SHIV一直是人们追求的目标。在这项提案中，我们的目标是使用一种新的方法来产生这样的SHIV。根据我们在操纵病毒序列方面的经验，我们优化了一种半自动化程序，该程序将允许通过大量R5-SHIV的感染性产生和筛选。此外，我们将使用最近被确定为在人类宿主中传播和建立感染的病毒的Env编码序列，这是潜在干预措施的精确目标。我们将测试SHIV在体外复制的能力。将混合有复制能力的病毒，并将使用病毒混合物对动物进行接种，并允许体内选择具有最高复制能力的病毒。最终，我们将产生与HIV-1毒株非常相似的SHIV，这些毒株在人类中传播，并在动物模型中有效复制。如果成功，这些研究将彻底改变艾滋病疗法和疫苗的临床前探索和开发。 公共卫生相关性：HIV-1是人类艾滋病的主要原因，它不能在大多数非人灵长类动物中复制，目前的动物模型也很有限。我们建议开发基于SIVMAC的新型嵌合病毒，该病毒表达与人类中流行的HIV-1毒株非常相似的HIV-1蛋白，并测试其作为猴HIV-1感染模型的实用性。如果成功，这一建议将导致改进HIV-1感染的动物模型，并将大大促进药物和疫苗干预措施的开发和测试。

DESCRIPTION (provided by applicant): HIV-1, the predominant cause of AIDS in humans, is unable to replicate in most non-human species. Therefore, the most practical animal model of AIDS consists of infection of rhesus macaques with SIVMAC or chimeras derived from SIVMAC that express the HIV-1 envelopes (SHIV). However, both of these models have specific limitations. In the case of SHIV, most chimeras developed to date use the CXCR4 co-receptor (X4-tropic) in contrast to most primary HIV-1 isolates that use the CCR5 co-receptor (R5-tropic). The search for R5-tropic SHIVs that replicate and cause disease in animals consistently has been much sought after goal. In this proposal we aim to use a novel approach towards the generation of such SHIVs. Based on our experience in manipulating viral sequences we have optimized a semi-automated procedure that will allow the generation and screening by infectivity of a large number of R5-SHIVs. Furthermore, we will use Env-coding sequences from viruses that have recently been identified as being transmitted and establishing infection in human hosts, the precise target of potential interventions. We will test the ability of our SHIVs to replicate in vitro. Replication competent viruses will be mixed and cocktails of viruses will be used to inoculate animals and allow in vivo selection of viruses with the highest replicative capacity. Ultimately, we will generate SHIVs that closely resemble HIV-1 strains that circulate in humans and replicate efficiently in an animal model. If successful, these studies should revolutionize the preclinical exploration and development of AIDS therapeutics and vaccines. PUBLIC HEALTH RELEVANCE: HIV-1, the predominant cause of AIDS in humans, is unable to replicate in most nonhuman primate species and current animal models are limited. We propose to develop novel chimeric viruses based on SIVMAC that express the HIV-1 proteins that closely resemble HIV-1 strains circulating in humans and to test their utility as an HIV-1 infection model in monkeys. If successful, this proposal will lead to improved animal models for HIV-1 infection and will considerably facilitate the development and testing of drug and vaccine interventions.