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

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，感染免疫系统的CD 4 + T细胞， 系统如果不及时治疗，HIV-1感染者将发展为艾滋病，并可能最终死亡。 联合抗逆转录病毒治疗在阻止感染者中HIV-1的复制方面非常有效。 个体尽管这种疗法成功地将HIV-1复制抑制到临床检测不到的水平， 抗逆转录病毒疗法不能治愈疾病。这是由于HIV-1一直处于沉默或潜伏状态 在称为静息记忆CD 4 + T细胞的CD 4 + T细胞亚群中。在这种潜伏状态下，这些被感染的细胞 抗逆转录病毒药物不能靶向它们，免疫系统也不能将其清除。HIV-1感染 在个体中，潜伏感染的CD 4 + T细胞的频率极低（约1/100万静息 记忆性CD 4 + T细胞），大多数在任何给定时间都存在于免疫组织中。这一人口 潜伏感染的细胞非常稳定，要求HIV-1感染者继续接受抗逆转录病毒治疗 以避免病毒血症反弹。因此，这群潜伏感染的CD 4 + T细胞是主要的免疫缺陷病毒。 治疗HIV-1感染的障碍。 开发消除潜伏感染细胞的策略是NIH、NIAID和NIH的主要焦点。 HIV-1研究领域。为了证明针对潜在水库的治疗方法的有效性，我们必须 能够使用快速和准确的测定来测量潜伏感染细胞的频率， 广泛的临床应用。重要的是，这种测定必须能够准确地测量潜在的细胞的大小。 病毒亚型的储存库。Zeroevir Diagnostics，LLC最近开发了IPDA作为一种新型的数字 液滴PCR测定，以测量外周血小样本中的完整和缺陷前病毒。IPDA 优化了用于B亚型HIV-1感染者，这种感染在美国占主导地位， 欧洲，但包括一小部分的人与艾滋病毒-1生活在世界各地。在这份提案中， 诊断寻求扩大IPDA的覆盖范围，包括C亚型HIV-1感染者， 约占全球HIV-1感染者的50%。总的来说，这项建议旨在 进行深入的前病毒测序，以告知分析覆盖范围的扩展，然后进行分析设计 适应，性能鉴定和分析的纵向收集的样本，从人的生活 C亚型HIV-1感染。该提案利用了与美国军方艾滋病研究中心的密切合作， 该计划使非洲各地的艾滋病毒1亚型C感染者能够获得独特的队列。

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.