Ongoing replication may occur on HAART

描述（由申请人提供）：hiv感染者积累了一个治疗抵抗库，潜伏感染的静息CD4+ T细胞，即使在接触后不久开始治疗。目前尚不清楚耐药HIV病毒库是如何维持的。有一种理论认为，潜伏感染的细胞在感染早期就已经形成，并且这些细胞在HAART治疗下仍然存在。第二种理论是，病毒的持久性需要非常低的HIV复制水平。此应用程序的目标是确定在HAART存在的情况下是否可以证明正在进行的复制。如果可以证明这种情况发生，则应将其视为水库维护的潜在机制。区分这些理论是很重要的，因为它将决定我们应该采取什么方法，如果要从受感染的患者中根除艾滋病毒感染。然而，无论储存库维持的机制或根除HIV的方法如何，检测正在进行的HIV复制的分析将有助于监测治疗。为了评估在HAART治疗的患者中是否发生持续的复制，我们建议在开始HAART治疗后一段时间内监测总HIV和整合HIV DNA。该提议背后的想法是，在没有持续复制的情况下，总DNA最终应该等于整合的HIV DNA。我们将测量血液和胃肠道中单个核细胞中的总和整合HIV DNA，并测试过量是否与正在进行的复制（Aim1A）的独立测量相关。此外，我们将测量这些细胞亚群（Aim1B）中的这些中间产物。最后，我们将采用一种补充方法，通过检测病毒向短寿命细胞（Aim1C）的传播来确定是否发生了持续的复制。我们期望我们的实验将提供关于正在进行的复制是否在HAART治疗的HIV感染者中发生的信息，并可能提供关于HIV发病机制的新信息。

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.