Measuring the Latent Reservoir and Monitoring Eradication Strategies

项目总结(见说明)： 在迄今开展的研究中，用于测量潜伏宿主的分析不准确，在某些情况下是非特异性的，在所有情况下都接近从有效接受抗逆转录病毒治疗的受试者获得的样本的检测阈值。合作实验室要想成功根除潜伏的油气藏，就必须有能够可靠地量化油气藏的化验方法，并且 衡量其规模的缩减。在项目4.1中，整合的病毒DNA和病毒感染性的高级分析方法将被改进和验证，用于艾滋病毒，然后应用于SIV和RT-Shiv猕猴模型。然后，它们将被应用于潜伏期的体外模型，以及从受感染的猕猴和艾滋病毒感染的研究对象获得的细胞和组织，作为其他合作实验室研究项目的一部分。 微流控技术在这些检测中的应用将解决具有复制能力的病毒的前病毒储存库的代表性问题，并试图解释在具有复制能力的病毒中复制的一些机制。还将检查作为潜在宿主的血液巨噬细胞中艾滋病毒DNA的存在和传染性。这些研究旨在改进对潜在储藏库的测量和清除它的干预措施，将涉及与项目和合作实验室核心调查人员的密切互动以及方法和材料的交流。

PROJECT SUMMARY (See instructions): The assays that have been applied to measure the latent reservoir in studies performed to date have been imprecise, in some cases non-specific, and in all cases near the threshold of detection for specimens obtained from subjects effectively treated with antiretroviral therapy. A successful Collaboratory effort to eradicate the latent reservoir must have assays available that can reliably quantify this reservoir and measure reductions in its size. In Project 4.1, superior assays in development for integrated viral DNA and for viral infectivity will be refined and validated for HIV, and then applied to both the SIV and RT-SHIV macaque models. They will then be applied to in vitro models of latency and to cells and tissues obtained from infected macaques and HIV-infected study subjects, as part of other Collaboratory research projects. Microfluidic applications to these assays will address the question of the representativeness of the proviral reservoir for replication-competent virus and try to explain some of the mechanisms for replication in competent virus. The presence of HIV DNA and infectivity in blood macrophages, as a potential reservoir, will also be examined. These studies to improve measurements of the latent reservoir and interventions to purge it will involve close interactions and the exchange of methods and materials with several of the project and core Collaboratory investigators.