Clinical Significance of HIV Replication Fitness

HIV复制适应度，我们将其定义为HIV分离株相对于参考HIV的生长速率 已提出体外培养的HIV-1菌株是HIV感染临床结果的重要预测因子。的 HIV复制适应性的临床意义仍然存在争议，主要是因为没有 在衡量相对复制适应度的最佳方法上达成了共识。关于适应性是否 使用从患者PBMC培养的全病毒的测定优于使用重组病毒的测定， 以及单循环测定法是否检测到与多循环生长竞争测定法相同的性质。 我们已经开发了一种重组病毒，多周期生长竞争试验来测量HIV复制 在细胞培养中的适应性。在该测定中，使用流式细胞术定量感染细胞的比例。我们 已经修改了该测定以测量病毒复制的单个循环期间的相对复制适合度，并且 还将修改生长竞争测定以测量完整（“整体”）临床细胞中的复制适合度。 分离株这些测定利用类似的病毒载体，并且将使用相同的方法定量病毒复制。 方法论然后，我们将在一个独特的位置，以评估如何这些健身测定比较， 确定这些方法中哪一种测量复制适合度与临床结果最相关。在 为了实现这些目标，我们将实现以下具体目标： (1)开发和评价一种多循环重组病毒检测方法，以测量 体外 (2)开发和评估单循环重组病毒检测方法，以测量体外HIV复制适应度。 (3)开发和评估一种多周期全病毒检测方法，以测量体外HIV复制适应度。 (4)确定使用这三种检测方法测量的HIV-1复制适应度是否与病毒感染相关。 治疗失败期间的病毒载量反弹或近期原发性HIV感染后的病毒载量设定点。 相关性：我们研究的总体目标是确定测量艾滋病毒感染速度的测试是否 可以预测HIV病毒载量水平。如果这些研究是成功的，他们可以确定测试， 帮助预测HIV感染者的临床结果。

HIV replication fitness, which we will define as the growth rate of an HIV isolate relative to a reference HIV strain in vitro, has been proposed to be an important predictor of clinical outcome in HIV infection. The clinical significance of HIV replication fitness continues to be controversial, primarily because there is no consensus on the best way to measure relative replication fitness. Arguments persist as to whether fitness assays, using whole virus cultured from patient PBMCs, are preferable to those using recombinant viruses, and whether single cycle assays detect the same properties as multiple-cycle growth competition assays. We have developed a recombinant virus, multiple-cycle growth competition assay to measure HIV replication fitness in cell culture. In this assay, the proportion of infected cells is quantified using flow cytometry. We have modified this assay to measure relative replication fitness during a single cycle of virus replication, and will also modify the growth competition assay to measure replication fitness in intact ("whole") clinical isolates. These assays utilize similar viral vectors, and will quantify viral replication using the same methodology. We will then be in a unique position to evaluate how these fitness assays compare, and determine which of these approaches to measure replication fitness best correlates with clinical outcome. In order to achieve these goals we will accomplish the following specific aims: (1) Develop and evaluate a multiple-cycle recombinant-virus assay to measure HIV replication fitness in vitro. (2) Develop and evaluate a single-cycle recombinant-virus assay to measure HIV replication fitness in vitro. (3) Develop and evaluate a multiple-cycle whole-virus assay to measure HIV replication fitness in vitro. (4) Determine whether HIV-1 replication fitness, as measured using these three assays, correlates with viral load rebound during treatment failure or viral load set point after recent primary HIV infection. Relevance: The overall goal of our studies is to determine whether tests that measure how rapidly HIV grows can predict HIV viral load level. If these studies are successful, they could identify tests that could help predict clinical outcomes for HIV-infected patients.