Falcinelli Shane D, Ceriani Cristina, Margolis David M, Archin Nancie M
UNC HIV Cure Center, The University of North Carolina at Chapel Hill, Chapel Hill, NC, United States.
Department of Microbiology and Immunology, The University of North Carolina at Chapel Hill, Chapel Hill, NC, United States.
Front Microbiol. 2019 Dec 18;10:2878. doi: 10.3389/fmicb.2019.02878. eCollection 2019.
A cure for HIV infection remains elusive due to the persistence of replication-competent HIV proviral DNA during suppressive antiretroviral therapy (ART). With the exception of rare elite or post-treatment controllers of viremia, withdrawal of ART invariably results in the rebound of viremia and progression of HIV disease. A thorough understanding of the reservoir is necessary to develop new strategies in order to reduce or eliminate the reservoir. However, there is significant heterogeneity in the sequence composition, genomic location, stability, and expression of the HIV reservoir both within and across individuals, and a majority of proviral sequences are replication-defective. These factors, and the low frequency of persistently infected cells in individuals on suppressive ART, make understanding the reservoir and its response to experimental reservoir reduction interventions challenging. Here, we review the characteristics of the HIV reservoir, state-of-the-art assays to measure and characterize the reservoir, and how these assays can be applied to accurately detect reductions in reservoir during efforts to develop a cure for HIV infection. In particular, we highlight recent advances in the development of direct measures of provirus, including intact proviral DNA assays and full-length HIV DNA sequencing with integration site analysis. We also focus on novel techniques to quantitate persistent and inducible HIV, including RNA sequencing and RNA/ protein staining techniques, as well as modified viral outgrowth methods that seek to improve upon throughput, sensitivity and dynamic range.
由于在抑制性抗逆转录病毒疗法（ART）期间具有复制能力的HIV前病毒DNA持续存在，HIV感染的治愈方法仍然难以捉摸。除了罕见的病毒血症精英控制者或治疗后控制者外，停止ART invariably会导致病毒血症反弹和HIV疾病进展。为了制定减少或消除病毒库的新策略，有必要全面了解病毒库。然而，HIV病毒库在个体内部和个体之间的序列组成、基因组位置、稳定性和表达方面存在显著异质性，并且大多数前病毒序列都是复制缺陷型的。这些因素，以及接受抑制性ART治疗的个体中持续感染细胞的频率较低，使得了解病毒库及其对实验性病毒库减少干预措施的反应具有挑战性。在这里，我们综述了HIV病毒库的特征、用于测量和表征病毒库的最新检测方法，以及在努力开发HIV感染治愈方法的过程中，这些检测方法如何能够准确检测病毒库的减少。特别是，我们强调了前病毒直接检测方法的最新进展，包括完整前病毒DNA检测和整合位点分析的全长HIV DNA测序。我们还关注定量持续性和可诱导性HIV的新技术，包括RNA测序和RNA/蛋白质染色技术，以及旨在提高通量、灵敏度和动态范围的改良病毒增殖方法。
