Development of a next-generation quantitative viral outgrowth assay (QVOA) for the standardized measurement of the HIV-1 latent reservoir

开发下一代定量病毒生长测定 (QVOA)，用于标准化测量 HIV-1 潜伏病毒库

• 批准号: 9346685
• 负责人: Gregory Michael Laird
• 金额: $ 29.72万
• 依托单位: ACCELEVIR DIAGNOSTICS, LLC
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-08-01 至 2018-10-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9346685
• 关键词: Acquired Immunodeficiency Syndrome; Address; Anti-Retroviral Agents; Antibodies; Automation; Benchmarking; Biological Assay; Biological Markers; Blood donor; CCR5 gene; CD4 Positive T Lymphocytes; Cell Cycle; Cell Line; Cell surface; Cells; Clinical; Clinical Trials; Collaborations; Cytogenetic Analysis; Development; Diagnostic; Ensure; Equipment; Frequencies; Funding Opportunities; Future; Goals; Gold; HIV-1; Immune system; Individual; Infection; Interruption; Laboratories; Laboratory Research; Left; Liquid substance; Logistics; Measurement; Measures; Methods; Mind; Mitogens; Monitor; National Institute of Allergy and Infectious Disease; Patients; Performance; Pharmaceutical Preparations; Phase; Phytohemagglutinins; Pilot Projects; Population; Procedures; Protocols documentation; Reagent; Reproducibility; Research; Resources; Rest; Retroviridae; Sampling; Scheme; Services; Small Business Innovation Research Grant; Standardization; Surface; Time; Training; Treatment Efficacy; United States National Institutes of Health; Universities; Validation; Viral; Virus; antiretroviral therapy; base; cell growth; clinically relevant; cost; drug development; improved; memory CD4 T lymphocyte; next generation; novel; prototype; reactivation from latency; reagent standard; success; targeted treatment; therapeutic target; validation studies; viral rebound

Project Summary/Abstract Human immunodeficiency virus type-1 (HIV-1) is a retrovirus that infects CD4+ T cells of the immune system. If left untreated, HIV-1 infected individuals will progress to AIDS and may ultimately die as a result. Combination antiretroviral therapy is extremely effective at stopping the replication of HIV-1 in infected individuals. Despite the success of this therapy at suppressing HIV-1 replication to clinically undetectable levels, antiretroviral therapy is not curative. This is due to the persistence of HIV-1 in a silent, or latent, state within a subset of CD4+ T cells known as resting memory CD4+ T cells. In this latent state, these infected cells are not targeted by antiretroviral drugs and cannot be eliminated by the immune system. In HIV-1 infected individuals, latently infected CD4+ T cells are found at extremely low frequencies (~1 per million resting memory CD4+ T cells). However, this population of latently infected cells is very stable, demanding that HIV-1 infected individuals remain on antiretroviral therapy indefinitely. Therefore, this population of latently infected CD4+ T cells is the main barrier to curing HIV-1 infection. Developing strategies to eliminate latently infected cells is a major focus of the NIH, NIAID, and the HIV-1 research field. To demonstrate the efficacy of therapeutics targeting the latent reservoir, we must be able to measure the frequency of latently infected cells using rapid and accurate assays that can be scaled for widespread clinical use. The currently accepted gold-standard assay for measuring latent HIV-1 is the quantitative viral outgrowth assay developed by the Siliciano laboratory in the mid-1990s. Due to the complexity and high resource requirements of the QVOA, this assay can only be performed in a small number of research laboratories. Accelevir Diagnostics, LLC is developing a new quantitative viral outgrowth assay with improved precision, reproducibility, and scalability. Broadly, this proposal aims to optimize assay conditions and perform key assay validation studies, including laboratory automation studies. The goal of this proposal is to develop an optimized commercial prototype, with accompanying standard operating procedures, and set the stage for rapid analytical validation and market entry. !

项目总结/摘要 人类免疫缺陷病毒1型（HIV-1）是一种逆转录病毒，感染CD 4 + T细胞， 免疫系统.如果不及时治疗，HIV-1感染者将发展为艾滋病， 最终导致死亡。联合抗逆转录病毒治疗在阻止 HIV-1在受感染个体中的复制。尽管这种治疗方法在 将HIV-1复制抑制到临床检测不到的水平，抗逆转录病毒疗法 有疗效的这是由于HIV-1在一个亚群中以沉默或潜伏状态持续存在， CD 4 + T细胞被称为静息记忆CD 4 + T细胞。在这种潜伏状态下，这些受感染的细胞 抗逆转录病毒药物不能靶向它，免疫系统也不能消除它。在HIV-1 感染个体中，潜伏感染的CD 4 + T细胞的频率极低（~1 每百万静息记忆CD 4 + T细胞）。然而，这群潜伏感染的细胞是 非常稳定，要求HIV-1感染者继续接受抗逆转录病毒治疗 无限期因此，这种潜伏感染的CD 4 + T细胞群体是免疫的主要屏障。 治愈HIV-1感染 开发消除潜伏感染细胞的策略是NIH的一个主要重点， NIAID和HIV-1研究领域。为了证明靶向治疗剂的疗效， 潜伏的水库，我们必须能够测量潜伏感染细胞的频率使用快速 和精确的分析，可以扩展到广泛的临床应用。目前公认的 用于测量潜伏HIV-1的金标准测定是定量病毒生长测定 由Siliciano实验室在20世纪90年代中期开发。由于复杂性和高资源 根据QVOA的要求，该试验只能在少量研究中进行 laboratories. Genevir Diagnostics，LLC正在开发一种新的定量病毒生长测定法， 改进的精度、再现性和可扩展性。概括地说，该建议旨在优化测定 条件下，并进行关键的分析验证研究，包括实验室自动化研究。 该提案的目标是开发一个优化的商业原型， 标准操作程序，并为快速分析验证和市场奠定基础 入境 !