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体外诱导静息 CD4+T 淋巴细胞分化增强病毒生长实验中复制型 HIV-1 的检测。

Ex Vivo Differentiation of Resting CD4+ T Lymphocytes Enhances Detection of Replication Competent HIV-1 in Viral Outgrowth Assays.

机构信息

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.

出版信息

Methods Mol Biol. 2022;2407:315-331. doi: 10.1007/978-1-0716-1871-4_21.

Abstract

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.

摘要

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。

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