HIV Dynamics and Replication Program, NCI, NIH, Frederick, MD 21702, USA.
Department of Biology, The Catholic University of America, Washington, DC 20064, USA.
Viruses. 2020 Jan 25;12(2):136. doi: 10.3390/v12020136.
Combination antiretroviral therapy (cART) controls but does not eradicate HIV infection; HIV persistence is the principal obstacle to curing infections. The proportion of defective proviruses increases during cART, but the dynamics of this process are not well understood, and a quantitative analysis of how the proviral landscape is reshaped after cART is initiated is critical to understanding how HIV persists. Here, we studied longitudinal samples from HIV infected individuals undergoing long term cART using multiplexed Droplet Digital PCR (ddPCR) approaches to quantify the proportion of deleted proviruses in lymphocytes. In most individuals undergoing cART, HIV proviruses that contain are lost more quickly than those that lack . Increases in the fraction of -deleted proviruses occurred only after 1-2 years of therapy, suggesting that the immune system, and/or toxicity of viral re-activation helps to gradually shape the proviral landscape. After 10-15 years on therapy, there were as many as 3.5-5 times more proviruses in which was deleted or highly defective than those containing intact . We developed a provirus-specific ddPCR approach to quantify individual clones. Investigation of a clone of cells containing a deleted HIV provirus integrated in the gene revealed that the cells underwent a massive expansion shortly after cART was initiated until the clone, which was primarily in effector memory cells, dominated the population of proviruses for over 6 years. The expansion of this HIV-infected clone had substantial effects on the overall proviral population.
联合抗逆转录病毒疗法（cART）可控制但不能根除 HIV 感染；HIV 持续存在是治愈感染的主要障碍。在 cART 期间，缺陷型前病毒的比例增加，但这一过程的动态尚不清楚，对 cART 启动后前病毒景观如何重塑进行定量分析对于了解 HIV 如何持续存在至关重要。在这里，我们使用多重数字 PCR（ddPCR）方法研究了接受长期 cART 的 HIV 感染者的纵向样本，以定量测定淋巴细胞中缺失前病毒的比例。在大多数接受 cART 的个体中，含有 的 HIV 前病毒比缺乏 的前病毒更快地丢失。 -缺失前病毒比例的增加仅在治疗 1-2 年后才发生，这表明免疫系统和/或病毒再激活的毒性有助于逐渐塑造前病毒景观。在接受治疗 10-15 年后，缺失或高度缺陷的 缺失或高度缺陷的前病毒比含有完整 的前病毒多 3.5-5 倍。我们开发了一种针对特定前病毒的 ddPCR 方法来定量个体克隆。对含有缺失 HIV 前病毒整合到 基因中的细胞克隆进行调查，结果表明，在 cART 开始后不久，这些细胞就经历了大规模扩增，直到该克隆（主要存在于效应记忆细胞中）在超过 6 年的时间里主导了前病毒群体。该 HIV 感染克隆的扩增对整个前病毒群体产生了重大影响。
