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

项目摘要 目前有超过3700万人感染艾滋病毒，每年仍有100多万人死于这种感染 年。尽管随着时间的推移，抗逆转录病毒疗法(ART)已经有了很大的进步，但它并不是治愈的。艾滋病病毒的屏障 CURE是静息的CD4+T细胞中潜伏的前病毒的储存库。如果患者停止抗逆转录病毒治疗，艾滋病毒也会 在几周内从这个蓄水池反弹，使疾病恶化。HIV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和 消灭他们。不幸的是，该领域还没有找到一种既能做到这一点又不会危及生命的药物。 因此，新的治疗策略必须在动物模型中进行测试：猴免疫缺陷病毒(SIV)和猿猴- 人类免疫缺陷病毒(SHIV)感染恒河猴。潜伏的艾滋病毒前病毒的情况是 最近发现的大部分是有缺陷的：只有2%的潜伏期前病毒完好无损。正因如此，聚合酶链式反应分析 放大前病毒基因组的一小部分给出了一个不准确的、多对数高估的潜在储存库。 目前，关于SIV和SIV模型中的潜油气藏的研究还很少。重要的是要确定 这些动物中的潜伏期前病毒池，以确定它们是否是 用来测试艾滋病毒的治愈策略。一旦了解了普罗鲁斯的情况，我们就必须设计新的分析方法来 精确测量SIV和SIV模型中的潜在储集层。我们唯一能确定我们的 治愈策略正在奏效，将疗法转移到临床试验是通过能够量化它们在我们的 动物模型。这项建议将表征潜伏的SIV和SHIV的景观，设计一部小说 只准确地测量复制能力强、完整的前病毒(真正的潜伏储存库)的化验，以及 用我们的新方法测试候选潜伏期反转剂对潜伏库的影响。

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.