Southern Research, Frederick, MD, USA.
Department of Pathology and Laboratory Medicine, Emory University School of Medicine, and Yerkes National Primate Research Center, Atlanta, GA, USA.
Methods Mol Biol. 2022;2407:315-331. doi: 10.1007/978-1-0716-1871-4_21.
Quantifying the number of cells harboring inducible and replication competent HIV-1 provirus is critical to evaluating HIV-1 cure interventions, but precise quantification of the latent reservoir has proven to be technically challenging. Existing protocols to quantify the frequency of replication-competent HIV-1 in resting CD4+ T cells from long-term ART treated individuals have helped to investigate the dynamics of reservoir stability, however these approaches have significant barriers to the induction of HIV-1 expression required to effectively evaluate the intact reservoir. Differentiation of CD4+ T cells to an effector memory phenotype is a successful strategy for promoting latency reversal in vitro, and significantly enhances the performance and sensitivity of viral outgrowth assays.
量化携带诱导和复制能力的 HIV-1 前病毒的细胞数量对于评估 HIV-1 治愈干预措施至关重要,但精确量化潜伏库一直是一项具有挑战性的技术难题。现有的从长期接受 ART 治疗的个体中静止 CD4+T 细胞中定量复制能力 HIV-1 的频率的方案有助于研究储库稳定性的动态,然而这些方法对于诱导 HIV-1 表达存在显著障碍,而这是有效评估完整储库所必需的。将 CD4+T 细胞分化为效应记忆表型是体外逆转潜伏的成功策略,可显著提高病毒扩增测定的性能和灵敏度。