Novel flow cytometry-based assay for HIV-infected cells

基于流式细胞术的新型 HIV 感染细胞检测方法

• 批准号: 8774585
• 负责人: JOSEPH M MCCUNE
• 金额: $ 21.06万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-12-01 至 2016-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8774585
• 关键词: Acquired Immunodeficiency Syndrome; Age; Biological Assay; Biological Models; Blood; CD4 Positive T Lymphocytes; Cardiovascular Diseases; Cell Count; Cell Line; Cells; Cellular Assay; DNA; Detection; Development; Disease; Disease Progression; Effectiveness; Epidemic; Evaluation; Event; Flow Cytometry; Fluorescent in Situ Hybridization; Frequencies; General Population; HIV; HIV Infections; Health; History of Medicine; In Vitro; Individual; Infection; Intervention Trial; Intestines; Investigation; Label; Location; Measurement; Measures; Memory; Methods; Modeling; Monitor; Neurocognitive Deficit; Nucleic Acids; Patients; Pattern; Phase; Phenotype; Population; Process; Production; RNA; Recovery; Resolution; Sampling; Sensitivity and Specificity; Signal Transduction; Specificity; Specimen; Subgroup; Testing; Time; Tissues; Viral; Virus; Virus Latency; Virus Replication; antiretroviral therapy; assay development; base; clinically relevant; cost; design; high throughput screening; in vitro Model; in vivo; killings; macrophage; novel; novel therapeutics; population based; prevent; reactivation from latency; research study; screening

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.

描述（由申请人提供）：HIV感染的抗逆转录病毒疗法（ART）是医学史上最重要，最快的进步之一。但是，尽管有效地限制病毒复制并减慢了疾病的进程，但如果没有病毒的快速反弹和疾病进展，就无法停止艺术。此外，尽管在ART上可以预防AIDS，但与一般人群相比，ART疾病和神经认知能力下降等重要的非AID事件（例如心血管疾病和神经认知下降）的发生频率更高，而在HIV感染的ART受试者的年龄较早。最后，流行病太大了，障碍 太高了，无法有效治疗每个受感染的人以结束流行病。因此，我们必须了解感染者如何持续的HIV如何设计新型疗法以消除病毒或在功能上治愈患者的机制。理解的第一步是开发测定病毒持久性水平的测定方法。存在几种测定法，但在提供的信息，可以执行的速度，有多昂贵以及测量的变化方面有严重的限制。我们在这里提出了一种新的方法，可以使用流式细胞仪在单细胞分辨率下检测HIV感染的细胞。一旦优化，该方法将提供更多信息，更快地执行，成本较小，并且比其他测量HIV储层的测定法更低。我们将优化该测定法，然后应用它以了解各种HIV感染的受试者中的HIV储层状态，以评估某些临床上不同的受试者是否可能具有不同的大小，活动或重新激活的艾滋病毒储备潜力，以开发出高通量筛选分析的高通量筛选分析，以评估新的治疗方法以对latenticals进行新的治疗或杀死latentsoivate seveceivate seveceivate seveceivate seveceivate seveDentosoIriair seveDentsoir。该过程将证明该测定法的实用性可以应用其应用，并传播其他人使用的测定法，以便在他们认为合适的情况下应用。