Clinical Significance of HIV Replication Fitness

描述(由申请人提供)：HIV复制适合度，我们将其定义为HIV分离株相对于参考HIV毒株的体外生长速度，已被认为是预测HIV感染临床结果的重要指标。艾滋病毒复制适合度的临床意义仍然存在争议，主要是因为对于衡量相对复制适合度的最佳方式还没有达成共识。对于使用从患者PBMC培养的全病毒的适合度分析是否比使用重组病毒的分析更可取，以及单周期分析是否检测到与多周期生长竞争分析相同的特性，争论仍在继续。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细胞培养中HIV复制的适合性。在这项检测中，感染细胞的比例是用流式细胞术来量化的。我们已经修改了这一检测方法，以测量病毒复制的单个周期中的相对复制适合性，并将修改生长竞争分析方法，以测量完整(“完整”)临床分离株的复制适合性。这些检测使用相似的病毒载体，并将使用相同的方法量化病毒复制。然后，我们将处于一个独特的位置来评估这些适应性分析如何比较，并确定这些方法中的哪些方法来衡量复制适应性与临床结果最相关。为了实现这些目标，我们将完成以下具体目标：(1)建立和评价多周期重组病毒体外复制适合性检测方法。(2)建立和评价单周期重组病毒体外复制适合性检测方法。(3)建立和评价多周期全病毒体外复制适合性检测方法。(4)确定使用这三种检测方法测量的HIV-1复制适合性是否与治疗失败期间的病毒载量反弹或最近首次感染艾滋病毒后的病毒载量设定点相关。相关性：我们研究的总体目标是确定衡量艾滋病毒增长速度的测试是否可以预测艾滋病毒病毒载量水平。如果这些研究成功，他们可以确定有助于预测艾滋病毒感染患者临床结果的测试。

DESCRIPTION (provided by applicant): 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.