Accurately Measuring the True Size of the Latent Reservoir and Characterizing the Proviral Landscape in SIV and SHIV Animal Models

项目摘要 目前有3 700多万人感染艾滋病毒，每年仍有100多万人死于这种感染。 年虽然抗逆转录病毒疗法（ART）随着时间的推移有了很大的改善，但它并不是治愈性的。艾滋病毒的障碍 cure是潜伏的前病毒的储存库，潜伏的前病毒隐藏在静止的CD4+ T细胞中。如果患者停止抗逆转录病毒疗法， 在几周内从这个水库反弹，允许疾病进展。艾滋病的治愈取决于 逆转含有前病毒的细胞的潜伏状态，从而允许免疫系统检测并 消灭他们。不幸的是，该领域还没有找到一种药物，可以做到这一点，而不会危及生命的毒性。 因此，新的治疗策略必须在动物模型中进行测试：猿免疫缺陷病毒（SIV）和猿免疫缺陷病毒（SIV）。 人免疫缺陷病毒（SHIV）感染恒河猴。潜伏的HIV前病毒的情况是 最近发现大多数是有缺陷的：只有2%的潜伏前病毒是完整的。正因为如此， 扩增前病毒基因组的一小部分给出了对潜在储库的不准确的、多个对数的高估。 目前，对SIV和SHIV模型中的潜在储层知之甚少。重要的是要确定 这些动物中潜伏前病毒的库，以确定它们是否是 来测试艾滋病治疗策略。一旦了解了前病毒的情况，我们必须设计新的检测方法， 准确测量SIV和SHIV模型中的潜在储层。我们唯一能确定 治疗策略正在发挥作用，并将治疗方法推向临床试验，是通过能够量化它们在我们 动物模型该提案将描述潜在的SIV和SIV前病毒的景观，设计一种新颖的 仅准确测量有复制能力的完整前病毒（真正的潜伏库）的测定，以及 使用我们的新测定法测试候选潜伏逆转剂对潜伏储库的影响。

Project Summary More than 37 million people are currently living with HIV, an infection that still kills over 1 million individuals per year. Although antiretroviral therapy (ART) has vastly improved over time, it is not curative. The barrier to HIV cure is a reservoir of latent proviruses harbored in resting CD4+ T cells. If a patient ceases ART, HIV will rebound from this reservoir within a matter of weeks, allowing disease progression. HIV cure depends on reversing the latent state of the cells containing provirus, thus allowing the immune system to detect and eliminate them. Unfortunately, the field has yet to find a drug that will do this without life-threatening toxicity. Thus, new cure strategies must be tested in animal models: simian immunodeficiency virus (SIV) and simian- human immunodeficiency virus (SHIV) infected rhesus macaques. The landscape of latent HIV proviruses was recently found to be mostly defective: only 2% of latent proviruses are intact. Because of this, PCR assays that amplify a small part of the proviral genome give an inaccurate, multiple-log overestimate of the latent reservoir. Currently, little is known about the latent reservoir in SIV and SHIV models. It is essential to characterize the pool of latent proviruses in these animals in order to determine whether they are accurate model systems in which to test HIV cure strategies. Once the landscape of proviruses is known, we must design novel assays to accurately measure the latent reservoir in SIV and SHIV models. The only way we can determine whether our cure strategies are working and move therapies to clinical trials is by being able to quantify their effect in our animal models. This proposal will characterize the landscape of latent SIV and SHIV proviruses, design a novel assay to accurately measure only the replication-competent, intact proviruses (the true latent reservoir), and test the effect of candidate latency reversing agents on the latent reservoir using our new assay.