Development of Intact Proviral DNA Assays for SIV and SHIV

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，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在体内持续处于沉默或潜伏状态， CD 4 + T细胞的一个亚群，称为静息记忆CD 4 + T细胞。在这种潜伏状态下， 抗逆转录病毒药物的靶向，不能被免疫系统消除。在HIV-1感染者中， 潜伏感染的CD 4 + T细胞的频率极低（约1/百万静息记忆CD 4 + T细胞）， 细胞），其中大多数在任何给定时间在免疫组织内发现。这些潜伏感染的细胞群 非常稳定，要求HIV-1感染者无限期地接受抗逆转录病毒治疗， 病毒血症反弹。因此，这群潜伏感染的CD 4 + T细胞是治愈HIV-1的主要障碍 感染 制定消除潜伏感染细胞的策略是NIH、NIAID和HIV-1的主要重点。 1研究领域SIV和/或SHIV感染的猕猴是研究HIV的关键非人灵长类动物模型。 1持久性和测试新的治疗方法。有效证明候选人的有效性 针对这些动物中潜在宿主的治疗方法，我们必须能够测量潜在的频率 使用快速和准确的检测方法，感染的细胞可以被广泛使用， 外周血和经处理的组织，并且可以转化为人类临床研究。塞韦维尔 因此，诊断有限责任公司正在与约翰霍普金斯医学院的Siliciano实验室合作， SIV和SHIV适应的完整前病毒DNA测定法（IPDA）和相应的对照用于这些关键的 动物研究。概括地说，该提案旨在为SIV IPDA开发和表征新型细胞系对照 和SIV IPDA，鉴定SIV IPDA和SIV IPDA的性能，并对 IPDA作为SIV/SHIV感染猕猴治疗性干预研究中的药效学生物标志物。

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. SIV and/or SHIV infected macaques are a critical non-human primate model for studying HIV- 1 persistence and testing novel therapeutic approaches. To effectively demonstrate the efficacy of candidate therapeutics targeting the latent reservoir in these animals, we must be able to measure the frequency of latently infected cells using rapid and accurate assays that can be scaled for widespread use, are amenable to analyzing both peripheral blood and processed tissues, and can be translated to human clinical studies. Accelevir Diagnostics, LLC is therefore working with the Siliciano lab at Johns Hopkins School of Medicine to developing SIV and SHIV adapted intact proviral DNA assays (IPDA) and corresponding controls for use in these critical animal studies. Broadly, the proposal aims to develop and characterize novel cell line controls for the SIV IPDA and SHIV IPDA, qualify the performance of the SIV IPDA and SHIV IPDA, and perform initial evaluation of the IPDA as a pharmacodynamic biomarker in SIV/SHIV infected macaque studies of curative interventions.