Overcoming Host Restriction Factors to Develop Better Animal Models for HIV/AIDS.

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，人类艾滋病最实用的动物模型是用SIVMAC感染恒河猴或由SIVMAC衍生的编码HIV-1包膜(SIV)的嵌合体。然而，由于HIV-1和SIVMAC是不同的病毒，这些模型的实用性受到限制。基于对过去几年中获得的物种特异性限制因素的了解，我们已经产生了一种重组病毒，命名为嗜猴HIV(StHIV)，它几乎完全来自HIV-1，但在体外可以在原代恒河猴和辫尾猕猴细胞中非常有效地复制。我们的初步数据显示，stHIV变种能够在体内以低水平在恒河猴中复制，在尾辫猕猴中高水平复制。这项提议的目的是产生具有更强的复制能力和在动物中引起艾滋病的改进的stHIV衍生物，并使用这些病毒来研究HIV-1在体内的发病机制。首先，我们将扩展我们最新的发现，即stHIV变种可以在尾辫猕猴中有效复制，并推导出这种病毒的分子克隆，该病毒在体内持续有效地在尾辫猕猴中复制。基于这种病毒，我们将产生R5嗜性变异体，并启动粘膜传播研究。其次，我们将通过产生对猕猴限制因子具有抵抗力的最小突变的HIV-1蛋白来减少stHIV构建物中存在的SIVMAC序列。第三，我们将确定SIVMAC蛋白在恒河猴体内复制stHIV中的作用。最后，我们将使用stHIV模型来评估HIV-1辅助蛋白在体内病毒复制中的作用。最终，我们将产生与HIV-1毒株非常相似的stHIV，这种毒株在人类中传播，并在动物模型中导致艾滋病。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。 与公共卫生相关：HIV-1是人类艾滋病的主要原因，无法在大多数非人类灵长类物种中复制，目前的动物模型有限。我们已经开发了基于HIV-1的新型嵌合病毒，并建议测试它们在猴子中作为HIV-1感染模型的实用性，并产生更多的HIV-1衍生病毒。如果成功，这项提议将导致改进HIV-1感染的动物模型，并将极大地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 human AIDS consists of infection of rhesus macaques with SIVMAC or chimeras derived from SIVMAC that encode the HIV-1 envelope (SHIV). However, the usefulness of these models is limited by the fact that HIV-1 and SIVMAC are distinct viruses. Based on an understanding of species-specific restriction factors that we have acquired during the past few years, we have generated a recombinant viruses, named simian tropic HIV (stHIV), that are almost entirely derived from HIV-1 but can replicate very efficiently in primary rhesus and pigtailed macaque cells in vitro. Our preliminary data show that stHIV variants are able to replicate at low levels in rhesus and high levels in pigtailed macaques in vivo. The aims of this proposal are to generate improved derivatives of stHIV with enhanced ability to replicate and cause AIDS in animals and to use these viruses to study HIV-1 pathogenesis in vivo. First, we will expand our very recent findings that an stHIV variant can replicate efficiently in pigtailed macaques and derive a molecular clone of this virus that consistently replicates efficiently in pigtailed macaques in vivo. Based on this virus we will generate R5-tropic variants and initiate mucosal transmission studies. Second, we will reduce the SIVMAC sequences present in stHIV constructs by generating minimally mutated HIV-1 proteins that can confer resistance to macaque restriction factors. Third, we will determine the role of SIVMAC proteins in stHIV replication in rhesus macaques. Finally, we will use the stHIV model to assess the role of HIV-1 accessory proteins in virus replication in vivo. Ultimately, we will generate stHIVs that closely resemble HIV-1 strains that circulate in humans and cause AIDS 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 have developed novel chimeric viruses based on HIV-1 and are proposing to test their utility as an HIV-1 infection model in monkeys and to generate additional HIV-1-derived viruses. 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.