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

摘要 HIV-1是人类艾滋病的主要原因，无法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感染SIVMAC或SIVMAC的猕猴 源自SIVMAC的嵌合体。然而，这些模型的实用性受到以下事实的限制：HIV-1和 SIVMAC是两种截然不同的病毒。基于对物种特定限制因素的理解，我们有 产生的猴嗜性艾滋病毒(StHIV)变异株几乎完全来自HIV-1，但可以在 扎着辫子的猕猴。在上一个资助周期中，我们利用动物适应技术开发了sthiv分离株。 会导致辫尾猕猴患上艾滋病。此外，我们还研究了已知限制因素对 并揭示了由干扰素α和干扰素诱导的新型抑制物的活性。 限制慢病毒在原代细胞中特定物种的复制。这项提议的目的是进一步 通过衍生新的病毒克隆来开发stHIV，这些病毒克隆在免疫完好的动物中始终是致病的。 我们还将根据在人类中传播的HIV-1毒株推导出stHIV变种，并最终得出 粘膜传播。为了实现这些目标，我们的研究将包括对病毒RNA的详细表征 体内适应病毒蛋白的剪接和功能特性。此外，我们将生成一个stHIV 变异体，其中100%的序列来自HIV-1毒株，通过设计HIV-1 Vif蛋白来获得 可以抵消病毒复制的一个主要物种特异性障碍：猕猴APOBEC3蛋白。最后，我们 目的寻找新型干扰素α诱导的抑制人类免疫缺陷病毒(StHIV)在猕猴细胞中复制并产生stHIV的药物 可以克服它们的变种。我们的初步数据表明，这些目标是可行的，并将导致 成功开发出具有所需特性的stHIV变异体，这一进展有可能 为HIV-1药物和疫苗开发改造非人类灵长类动物模型。

ABSTRACT 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 macaques with SIVMAC or chimeras derived from SIVMAC. 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, we have generated simian tropic HIV (stHIV) variants that are almost entirely derived from HIV-1 but can replicate in pigtailed macaques. During the last funding cycle, we have used animal adaptation to develop stHIV isolates that cause AIDS in pigtail macaques. Additionally, we have studied the effects of known restriction factors on stHIV replication and unveiled the activity of novel, as yet unidentified, inhibitors that are induced by IFNα and limit lentiviral replication in a species-specific manner in primary cells. The aims of this proposal are to further develop stHIV by deriving novel viral clones that are consistently pathogenic in immunologically intact animals. We will also derive stHIV variants based on HIV-1 strains circulating in humans and ultimately variants for mucosal transmissions. To achieve these aims our studies will include a detailed characterization of viral RNA splicing and functional characterization of in vivo-adapted viral proteins. Additionally, we will generate an stHIV variant where 100% of the sequence is derived from HIV-1 strains by engineering an HIV-1 Vif proteins that can counteract a major species-specific barrier to virus replication: macaque APOBEC3 proteins. Finally, we aim to identify novel IFNα-induced inhibitors that limit stHIV replication in macaque cells and generate stHIV variants that can overcome them. Our preliminary data suggest that these goals are feasible and will lead to the successful development of stHIV variants with the desired properties, an advance that has the potential to transform non-human primate models for HIV-1 drug and vaccine development.