Ongoing replication may occur on HAART

描述(由申请人提供)：即使在暴露后不久开始治疗，艾滋病毒感染者也积累了一个耐药的、潜伏感染的静息CD4+T细胞的储存库。目前尚不清楚耐药艾滋病毒病毒库是如何维持的。一种理论认为，潜伏感染细胞是在感染早期建立的，这些细胞在HAART存在的情况下持续存在。第二种理论认为，病毒的持久性需要非常低的艾滋病毒复制水平。这个应用程序的目标是确定是否可以证明在存在HAART的情况下发生正在进行的复制。如果能够证明这一点，那么它应该被认为是一种潜在的储油层维护机制。区分这些理论很重要，因为它将推动我们应该采取什么方法，如果要从感染患者中根除艾滋病毒感染。然而，无论水库维持的机制(S)或根除艾滋病毒的方法如何，检测正在进行的艾滋病毒复制的分析将对监测治疗有用。为了评估接受HAART的患者是否发生正在进行的复制，我们建议在开始HAART后随着时间的推移监测总的和整合的HIV DNA。这项提议背后的想法是，在没有持续复制的情况下，总DNA最终应该等于整合的艾滋病毒DNA。我们将测量血液和胃肠道中单个核细胞中总的和整合的HIV DNA，并测试过量是否与正在进行的复制的独立测量相关(Aim1A)。此外，我们还将测量这些细胞亚群中的中间产物(Aim1B)。最后，我们将采取一种补充方法，通过测试病毒传播到短期细胞(Aim1C)来确定是否发生正在进行的复制。我们希望我们的实验将提供关于HAART中HIV感染者是否发生正在进行的复制的信息，并可能提供关于HIV发病机制的新信息。 与公共卫生相关：尚不清楚为什么艾滋病毒是可以治疗的，但不是可以治愈的。似乎有一个耐治疗的储存库，但在HAART存在的情况下如何维持这个储存库尚不清楚。在这份赠款提案中，我们试图通过确定抗病毒治疗是否完全阻止艾滋病毒复制来解决这个问题。我们提出的实验试图通过测量抗病毒治疗中随时间推移的病毒DNA中间体来回答这个问题。

DESCRIPTION (provided by applicant): HIV-infected individuals accumulate a reservoir of treatment-resistant, latently infected resting CD4+ T cells, even when therapy is started shortly after exposure. It remains unclear how treatment resistant HIV viral reservoirs are maintained. One theory is that latently infected cells are established early in infection and that these cells persist in the presence of HAART. A second theory is that a very low level of HIV replication is required for viral persistence. The goal of this application is to determine if ongoing replication can be demonstrated to occur in the presence of HAART. If this can be demonstrated to occur then it should be considered as a potential mechanism for reservoir maintenance. Distinguishing between these theories is important because it will drive what approaches we should take if HIV infection is to be eradicated from infected patients. However, regardless of the mechanism(s) of reservoir maintenance or the approaches to eradication of HIV, assays that detect ongoing HIV replication would be useful for monitoring therapy. In an effort to assess if ongoing replication occurs in patients on HAART, we propose to monitor total and integrated HIV DNA over time after initiating HAART. The idea behind the proposal is that in the absence of ongoing replication total DNA should eventually equal integrated HIV DNA. We will measure total and integrated HIV DNA in mononuclear cells in both blood and the GI tract and test if an excess correlates with independent measures of ongoing replication (Aim1A). In addition, we will measure these intermediates within subsets of these cells (Aim1B). Finally, we will take a complementary approach to determine if ongoing replication occurs by testing for viral spread to short lived cells (Aim1C). We expect our experiments will provide information on whether ongoing replication occurs in HIV infected individuals on HAART and potentially new information on HIV pathogenesis. PUBLIC HEALTH RELEVANCE: It is unclear why HIV is treatable, but not curable. It appears that there is a treatment resistant reservoir, but how this reservoir is maintained in the presence of HAART is unclear. In this grant proposal, we try to address this question by determining if HIV replication is completely stopped with antiviral therapy. We propose experiments that attempt to answer this question by measuring viral DNA intermediates over time on antiviral therapy.