IPDA for High-Priority HIV-1 Subtype C to Enable Global Eradication Trials

项目摘要/摘要 人类免疫缺陷病毒1型(HIV-1)是一种逆转录病毒，感染免疫的CD4+T细胞 系统。如果不进行治疗，HIV-1感染者将发展为艾滋病，并可能最终死亡。 联合抗逆转录病毒疗法在阻止HIV-1在感染者体内复制方面极其有效 个人。尽管这种疗法成功地将HIV-1复制抑制到临床无法检测到 水平，抗逆转录病毒治疗是不能治愈的。这是由于HIV-1持续处于沉默或潜伏状态 在被称为静息记忆的CD4+T细胞的子集内。在这种潜伏状态下，这些被感染的细胞 不是抗逆转录病毒药物的靶点，也不能被免疫系统消除。在感染了HIV-1的人中 在个体中，潜伏感染的CD4+T细胞被发现的频率极低(每百万人中有1人处于静息状态 记忆中的CD4+T细胞)，在任何给定的时间，大多数存在于免疫组织中。这一人口 潜伏感染细胞非常稳定，要求HIV-1感染者继续接受抗逆转录病毒治疗 无限期避免病毒血症反弹。因此，这种潜伏感染的CD4+T细胞是主要的 治愈HIV-1感染的障碍。 开发消除潜伏感染细胞的策略是NIH、NIAID和 HIV-1研究领域。为了证明针对潜在储蓄者的疗法的有效性，我们必须 能够使用快速而准确的分析方法测量潜伏感染细胞的频率，该方法可根据需要进行调整 广泛的临床应用。关键是，这样的分析必须能够准确地测量潜伏期的大小。 病毒亚型的储存库。Acelevir Diagnostics，LLC最近开发了IPDA作为一种新的数字 滴状聚合酶链式反应检测小样本外周血中完整和缺陷前病毒。IPDA 被优化用于感染B型HIV-1亚型的人，这种亚型在美国和 欧洲的艾滋病毒携带者只占全球艾滋病毒携带者的一小部分。在这份提案中，阿科雷维尔 诊断寻求扩大IPDA的覆盖范围，以包括感染C型艾滋病毒-1亚型的人，这 约占全球艾滋病毒携带者总数的50%。总的来说，这项提议旨在 执行深入的前病毒测序，以通知分析覆盖范围的扩展，然后进行分析设计 纵向采集的精神分裂症患者样本的适应、行为鉴定和分析 C亚型HIV-1感染。这项提议利用了与美国军方艾滋病毒研究中心的密切合作 该计划使非洲能够接触到独一无二的C亚型HIV-1感染者。

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), with the majority found within immune tissues at any given time. This population of latently infected cells is very stable, demanding that HIV-1 infected individuals remain on antiretroviral therapy indefinitely to avoid rebound of viremia. As such, 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. Critically, such assays must be capable of accurately measuring the size of the latent reservoir across viral subtypes. Accelevir Diagnostics, LLC has recently developed the IPDA as a novel digital droplet PCR assay to measure intact and defective proviruses in a small sample of peripheral blood. The IPDA was optimized for use in people with subtype B HIV-1 infection, which predominates in the United States and Europe but comprises on a small fraction of people living with HIV-1 worldwide. In this proposal, Accelevir Diagnostics seeks to expand the IPDA coverage to include people living with subtype C HIV-1 infection, which accounts for approximately 50% of all people living with HIV-1 worldwide. Broadly, this proposal aims to perform in-depth proviral sequencing to inform expansion of assay coverage followed by assay design adaptation, performance qualification, and analysis of longitudinally collected samples from people living with subtype C HIV-1 infection. This proposal leverages a close collaboration with the US Military HIV Research Program, enabling access to a unique and cohort of people living with subtype C HIV-1 infection across Africa.