HIV/SIV viral reservoirs in lymphocytes and macrophages

拟议的研究涉及HIV-1在接受高效抗逆转录病毒治疗（HAART）的患者中持续存在的机制。在依从性患者中，HAART可以将血浆中的游离病毒水平降低到检测极限以下。然而，在最初资助期间完成的研究表明，HIV-1在一小部分潜伏感染的静止记忆CD4 + T细胞中持续存在，这些细胞携带HIV-1基因组的完整拷贝。由于潜伏感染的细胞是消灭HIV-1的主要障碍，因此了解维持潜伏的分子机制非常重要。基于体外研究，已经提出了几种潜在的分子机制，但尚不清楚这些机制是否能准确地解释HIV-1在体内静止的CD4 + T细胞中的持久性。在拟议的研究中，我们将使用遗传学方法来研究体内潜伏期的机制，重点关注

The proposed studies deal with the mechanism by which HIV-1 persists in patients on highly active antiretroviral therapy (HAART). In compliant patients, HAART can decrease levels of free virus in the plasma to below the limit of detection. However, studies completed during the initial funding period demonstrated that HIV-1 persists for life in a small pool of latently infected resting memory CD4 + T cells that carry an integrated copy of the HIV-1 genome. Because latently infected cells represent a major barrier to HIV-1 eradication, it is important to understand the molecular mechanisms that maintain latency. Several potential molecular mechanisms have been proposed based on in vitro studies, but it remains unclear whether any of these accurately account for the persistence of HIV-1 in resting CD4 + T cells in vivo. In the proposed studies, we will use genetic approaches to examine the mechanism of latency in vivo, focusing on the influence of the integration site in the host genome. The first two Specific Aims involve cloning and characterizing sites of HIV-1 integration in resting CD4 + T cells in vivo. A novel method will be used to identify the small subset of integrated proviruses that are functionally significant. Analysis of these integration sites should provide a general picture the chromosomal environments where latent proviruses reside. In the third Specific Aim, we will use integration site information to study the transcriptional activity and chromatin structure of regions of the host chromosome that are sites for HIV-1 integration. In the final Aim, we will test a novel hypothesis concerning the mechanism of HIV-1 latency that is consistent with most published work and that represents a synthesis of several proposed mechanisms. Together, these studies should help to elucidate how HIV-1 latency operates in vivo.