Testing of VIF Inhibitors Against Clinical Isolates and Animal Study Support

核心B将通过提供有希望的Vif的特定体外和体内测试来支持整体拨款申请 抑制剂候选物。第一个目的是测试有希望的Vif抑制剂候选物在治疗中的功效和毒性。 使用临床HIV-1分离株（包括嗜神经性分离株）的基于人PBMC的测定。第二个目标是 在HIV-1中空纤维小鼠模型中测试有希望的Vif抑制剂候选物。中空纤维填充有 将HIV-1感染的细胞皮下和腹膜内植入Balb/c小鼠。随后，委员会注意到， 将用Vif抑制剂候选物腹膜内处理小鼠。将监测小鼠的 急性毒性和评价纤维对HIV-1复制的功效。第三个目标是执行 恒河猴血浆和病毒载量样品的定量RNA病毒载量测定，以支持 新英格兰灵长类研究中心的SIV/灵长类研究（项目3）。对于定量病毒载量 为了进行测量，将使用ABI 7900测序仪进行TAQMAN RT-PCR。引物、探针和 标准品来自SIV的gag区。 该项目与公共卫生的相关性： 目前，美国约有100万人感染艾滋病毒。此外，4万名新 每年都有感染发生。到目前为止，约有50万人死于艾滋病毒感染并发症 在美国.在过去的15年里，已经开发了多种抗HIV化合物，但这些化合物 只能控制但不能治愈感染。目前获得许可的所有化合物都是针对两种 HIV酶或针对病毒/细胞融合。最近，另一种HIV蛋白Vif与细胞内的 蛋白（APOBEC 3G）。新发现的抑制这种相互作用的化合物具有 由于Vif是在原代人类细胞中复制所必需的，因此具有治愈HIV-1感染的潜力。

Core B will support the overall grant application by providing specific in vitro and in vivo tests of promising Vif inhibitor candidates. The first aim is to test the efficacy and toxicity of promising Vif inhibitor candidates in human PBMC-based assays using clinical HIV-1 isolates including neurotropic isolates. The second aim is to test promising Vif inhibitor candidates in the HIV-1 Hollow Fiber Mouse model. Hollow fibers filled with HIV-1 infected cells will be implanted subcutaneously and intraperitoneally into Balb/c mice. Subsequently, the mice will be treated intraperitoneally with the Vif inhibitor candidates. The mice will be monitored for acute toxicity and the fibers evaluated for efficacy against HIV-1 replication. The third aim is to perform quantitative RNA viral load determinations on rhesus macaque plasma and viral load samples in support of the SIV/primate studies at New England Primate Research Center (Project 3). For the quantitative viral load measurements, TAQMAN RT-PCR using an ABI7900 sequencer will be performed. Primers, probe, and standard are from the gag region of SIV. The relevance of this project to public health: Currently, about 1 million people are infected with HIV in the United States. In addition, 40,000 new infections are occurring each year. So far, about 500,000 persons died from complications of HIV-infection in the US. In the last 15 years, a variety of anti-HIV compounds have been developed but these compounds only control but do not cure the infection. All of the compounds currently licensed are directed against two HIV enzymes or against virus/cell fusion. Recently, the interaction of another HIV protein, Vif, with a cellular protein (APOBEC3G) was discovered. The newly identified compounds that inhibit this interaction have the potential to cure the infection with HIV-1 since Vif is required for replication in primary human cells.