Characterizing the viral and host effector mechanisms that govern HIV-1 rebound

项目摘要 HIV/AIDS治疗研究的一个中心目标是预防或延迟分析治疗后的病毒反弹 中断（ATI）。人们普遍认为，了解病毒和宿主因素参与体内潜伏期 重新激活将导致新的，更合理的治疗策略;然而， 反弹病毒仍然是未知的。在这里，我们建议利用我们最近的四个发现， 小组深入了解HIV-1复发的机制。采用“黄金标准”抽样方法 为了表征治疗中断（ATI）前后的潜在储库，我们发现病毒分离株 通过定量病毒生长（QVOA）得出的结果并不代表或预测血浆中出现的病毒 治疗中断后（1，2）。我们还发现，反弹病毒，但不是QVOA衍生水库 病毒，对1型干扰素（IFN-I）具有高度抗性，表明宿主在干扰素受体的位点具有强有力的先天性应答。 病毒复发（3）。令人惊讶的是，有些人，但不是全部，反弹 表明 分离物在巨噬细胞中复制到高滴度， 比以前所知的更丰富的生物多样性。最后，我们发现， 反弹病毒有能力重新播种水库，提高了长期临床后果的可能性 ATI（2，3）。在本申请中，我们将利用这些发现来阐明病毒和宿主因素 控制HIV-1反弹的基因我们的假设是，通过（i）确定普遍性，动力学和临床 IFN-I抗性在反弹期间的影响，（ii）定义IFN-I抗性的病毒决定因素和宿主 干扰素刺激基因（ISG），对反弹的病毒施加压力，和（iii）追踪来源 IFN-I抗性反弹病毒，我们将揭示控制HIV-1从潜伏期再激活的关键机制。 在目标1中，我们将把研究扩展到更多样化的ATI试验参与者，包括女性、少数民族、急性 以及早期ART启动者和接受IFN-I调节疗法的个体，以评估 反弹病毒的IFN-I抗性表型。我们还将测试在何种程度上IFN-I抵抗持续， 延长ATI，并确定这如何影响IFN-I抗性病毒重新播种水库的速度。在 目的2，我们将阐明反弹的HIV-1的生物学特性，绘制IFN-I抗性的病毒决定因素， 并确定宿主干扰素刺激基因（ISG）对复发病毒施加压力。在目标3中， 我们将通过检测血液、胸导管淋巴液和淋巴组织来追踪反弹病毒的来源， 定期和修改的QVOA，检查是否长寿命的骨髓细胞，包括脑巨噬细胞， 小胶质细胞，作为IFN-I抗性病毒的储存库，并探索是否SHIV感染的恒河猴 概括HIV-1储库和反弹病毒的IFN-Ⅰ表型。我们希望这些研究能有所改进 我们对临床相关的、有反弹能力的HIV-1储库的理解， 体内病毒再激活和宿主因素对反弹的病毒库施加压力，这应该导致 更合理有效的艾滋病治疗策略。 .

PROJECT SUMMARY A central goal in HIV/AIDS cure research is preventing or delaying viral rebound following analytical treatment interruption (ATI). It is widely assumed that understanding the viral and host factors involved in in vivo latency reactivation would lead to new, more rational cure strategies; however, the biology and provenance of the rebounding virus remains largely unknown. Here, we propose to leverage four recent discoveries from our groups to gain insight into the mechanisms of HIV-1 recrudescence. Using “gold standard” sampling methods to characterize the latent reservoir before and after treatment interruption (ATI), we discovered that viral isolates derived by quantitative virus outgrowth (QVOA) do not represent or predict the viruses that emerge in the plasma following treatment interruption (1, 2). We also found that rebound viruses, but not QVOA-derived reservoir viruses, were highly resistant to type 1 interferons (IFN-I), indicating potent host innate responses at the site of viral recrudescence (3). Surprisingly, some, but not all, rebound suggesting isolates replicated to high titers in macrophages, a greater biological diversity than previously known. Finally, we discovered that IFN-I resistant rebound viruses have the ability to reseed the reservoir, raising the possibility of long-term clinical consequences of ATI (2, 3). In this application, we will leverage these discoveries to elucidate the viral and host factors that govern HIV-1 rebound. Our hypothesis is that by (i) determining the universality, kinetics and clinical impact of IFN-I resistance during rebound, (ii) defining the viral determinants of IFN-I resistance and the host interferon stimulated genes (ISGs) that place pressure on the rebounding virus, and (iii) tracing the provenance of IFN-I resistant rebound viruses, we will uncover key mechanisms that control HIV-1 reactivation from latency. In Aim 1, we will expand our studies to more diverse ATI trial participants, including women, minorities, acute and early ART initiators and individuals receiving IFN-I modulating therapies, to assess the generality of the IFN-I resistant phenotype of rebound viruses. We will also test to what extent IFN-I resistance persists during prolonged ATI and determine how this influences the rates of IFN-I resistant viruses that reseed the reservoir. In Aim 2, we will elucidate the biological properties of rebound HIV-1, map the viral determinants of IFN-I resistance, and identify the host interferon stimulated genes (ISGs) that place pressure on the recrudescing virus. In Aim 3, we will trace the provenance of rebound virus by testing blood, thoracic duct lymph and lymphatic tissue using regular and modified QVOAs, examine whether long-lived myeloid cells, including brain macrophages and microglia, serve as reservoirs of IFN-I resistant viruses, and explore whether SHIV-infected rhesus macaques recapitulate the IFN-I phenotypes of HIV-1 reservoir and rebound viruses. We expect these studies to improve our understanding of the clinically relevant, rebound competent HIV-1 reservoir, the mechanisms underlying in vivo viral reactivation and the host factors that place pressure on the rebounding virus pool, which should lead to more rational and effective HIV/AIDS cure strategies. .