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. !

项目概要/摘要 人类免疫缺陷病毒 1 型 (HIV-1) 是一种逆转录病毒，可感染 CD4+ 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，并可能 最终因此而死亡。联合抗逆转录病毒疗法对于阻止病毒极其有效 HIV-1 在感染者体内的复制。尽管这种疗法在 抗逆转录病毒治疗不能抑制 HIV-1 复制至临床无法检测的水平 有疗效。这是由于 HIV-1 在一个子集内持续处于沉默或潜伏状态。 CD4+ T 细胞称为静息记忆 CD4+ T 细胞。在这种潜伏状态下，这些受感染的细胞 不是抗逆转录病毒药物的目标，也不能被免疫系统消除。 HIV-1中 在感染个体中，潜伏感染的 CD4+ T 细胞的发现频率极低（~1 每百万静息记忆 CD4+ 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 无限期地。因此，潜伏感染的 CD4+ T 细胞群是感染的主要障碍。 治愈 HIV-1 感染。 制定消除潜伏感染细胞的策略是 NIH 的主要重点， NIAID 和 HIV-1 研究领域。为了证明靶向治疗的功效 潜伏病毒库，我们必须能够使用快速测量潜伏感染细胞的频率 以及可扩展以供广泛临床使用的准确测定。目前接受的 测量潜伏 HIV-1 的金标准测定是定量病毒生长测定 由 Siliciano 实验室于 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，该测定只能在少数研究中进行 实验室。 Accelevir Diagnostics, LLC 正在开发一种新的定量病毒生长检测方法 提高了精度、再现性和可扩展性。总的来说，该提案旨在优化检测 条件并进行关键的测定验证研究，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，并附带 标准操作程序，并为快速分析验证和市场奠定基础 入口。 ！