Novel flow cytometry-based assay for HIV-infected cells

描述（由申请人提供）：针对HIV感染的抗逆转录病毒疗法（ART）是医学史上最重要和最迅速的进步之一。然而，尽管抗逆转录病毒疗法能有效地限制病毒复制和减缓疾病进程，但如果病毒不迅速反弹和疾病不迅速进展，就无法停止抗逆转录病毒疗法。此外，虽然在接受抗逆转录病毒治疗的患者中可以预防艾滋病，但在接受抗逆转录病毒治疗的艾滋病毒感染者中，心血管疾病和神经认知能力下降等重大非艾滋病事件的发生频率和年龄比一般人群要高。最后，疫情规模太大，开展工作存在障碍

DESCRIPTION (provided by applicant): Antiretroviral therapy (ART) for HIV infection is one of the most important and rapid advances in the history of medicine. However, while effective at limiting virus replication and slowing the course of disease, ART cannot be stopped without rapid rebound of virus and progression of disease. In addition, while AIDS is prevented in patients on ART, significant non-AIDS events such as cardiovascular disease and neurocognitive decline occur more frequently and at an earlier age in HIV-infected subjects on ART than in the general population. Finally, the epidemic is too large and the barriers to roll-out too high to effectively treat every infected individual to end the epidemic. It is therefore imperative that we understand the mechanisms of how HIV persists in the infected individual to design novel therapies to eradicate the virus or functionally cure the patient. The first step in tis understanding is the development of assays to measure the level of virus persistence. Several assays exist, but have serious limitations in terms of how much information is provided, how quickly they can be performed, how expensive they are, and how variable the measurement is. We propose here a novel method to detect HIV-infected cells at single cell resolution using flow cytometry. This method, once optimized, will provide more information, be performed more rapidly, cost less, and have less variability than other assays for the measurement of HIV reservoirs. We will optimize this assay and then apply it to understand the state of the HIV reservoir in various groups of HIV-infected subjects to evaluate whether some clinically different subjects may have different sizes, activities, or reactivation potentials of their HIV reservoirs ad also to develop a high-throughput screening assay for the evaluation of novel therapeutics to reactivate or kill the latent reservoir. This process will demonstrate the utility of this assay an highlight situations in which it can be applied as well as disseminate the assay for use by others to apply as they see fit.