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

描述(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，最实用的艾滋病动物模型是用SIVMAC感染恒河猴或由SIVMAC衍生的表达HIV-1包膜(SIV)的嵌合体。然而，这两种模式都有特定的局限性。在SHIV的病例中，迄今为止开发的大多数嵌合体使用CXCR4共受体(X4嗜性)，而大多数初级HIV-1分离株使用CCR5共受体(R5嗜性)。寻找在动物身上持续复制和致病的R5嗜性SHIVE一直是人们追逐的目标。在这项提议中，我们的目标是使用一种新的方法来产生这种切割器。根据我们在操纵病毒序列方面的经验，我们已经优化了一种半自动程序，该程序将允许通过感染性产生和筛选大量R5-SHIV。此外，我们将使用最近被确认为正在传播的病毒的环境编码序列，并确定在人类宿主中的感染，这是潜在干预的确切目标。我们将测试我们的SHIV在体外复制的能力。具有复制能力的病毒将混合在一起，鸡尾酒病毒将用于接种动物，并允许在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，这种毒株在人类中传播，并在动物模型中高效复制。如果成功，这些研究将彻底改变艾滋病治疗和疫苗的临床前探索和发展。

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.