Department of Medicine, Duke Universitygrid.26009.3d School of Medicine, Durham, North Carolina, USA.
Duke Human Vaccine Institute, Duke Universitygrid.26009.3d School of Medicine, Durham, North Carolina, USA.
J Virol. 2022 Jul 27;96(14):e0062422. doi: 10.1128/jvi.00624-22. Epub 2022 Jul 5.
HIV-1 persistence in different cell types presents the main obstacle to an HIV-1 cure. We have previously shown that the renal epithelium is a site of HIV-1 infection and that the kidney represents a separate viral compartment from blood. Whether renal cells can harbor latent virus that can be reactivated upon treatment with latency reversing agents (LRAs) is unknown. To address this question, we developed an HIV-1 latency model in renal tubule epithelial (RTE) cells using a dual color HIV-1 reporter virus, R7/E-/GFP/EF1a-mCherry (R7GEmC), and evaluated the effect of LRAs, both as single agents and in combination, on viral reactivation. Our data show that HIV-1 can establish latency in RTE cells early postinfection. While the pool of latently infected cells expanded overtime, the percentage of productively infected cells declined. Following LRA treatment only a small fraction of latently infected cells, both T cells and RTE cells, could be reactivated, and the drug combinations more effective in reactivating HIV transcription in RTE cells differed from those more active in T cells. Our study demonstrates that HIV can establish latency in RTE cells and that current LRAs are only marginally effective in inducing HIV-1 reactivation. This suggests that further study of LRA dynamics in non-T cells may be warranted to assess the suitability of LRAs as a sterilizing cure strategy. Anti-retroviral therapy (ART) has dramatically reduced HIV-related morbidity and mortality. Despite this success, a number of challenges remain, including the long-term persistence of multiple, clinically latent viral reservoirs capable of reactivation in the absence of ART. As efforts proceed toward HIV eradication or functional cure, further understanding of the dynamics of HIV-1 replication, establishment of latency and mechanisms of reactivation in reservoirs harboring the virus throughout the body is necessary. HIV-1 can infect renal epithelial cells and the expression of viral genes in those cells contributes to the development of HIV associated nephropathy (HIVAN) in untreated individuals. The significance of our work is in developing the first model of HIV-1 latency in renal epithelial cells. This model enhances our understanding of HIV-1 latency and persistence in the kidney and can be used to screen candidate latency reversing agents.
HIV-1 在不同细胞类型中的持续存在是实现 HIV-1 治愈的主要障碍。我们之前已经表明，肾脏上皮细胞是 HIV-1 感染的部位，并且肾脏代表与血液不同的病毒隔室。尚不清楚肾脏细胞是否可以携带潜伏病毒，这些病毒可以在使用潜伏逆转剂 (LRA) 治疗后被重新激活。为了解决这个问题，我们使用双荧光素酶 HIV-1 报告病毒 R7/E-/GFP/EF1a-mCherry (R7GEmC) 在肾小管上皮 (RTE) 细胞中开发了 HIV-1 潜伏模型，并评估了 LRA 作为单一药物以及联合使用对病毒再激活的影响。我们的数据表明，HIV-1 可以在感染后早期在 RTE 细胞中建立潜伏。虽然潜伏感染细胞的池随着时间的推移而扩大，但产感染细胞的百分比下降。在用 LRA 治疗后，只有一小部分潜伏感染的细胞，包括 T 细胞和 RTE 细胞，可以被重新激活，并且在 RTE 细胞中更有效地重新激活 HIV 转录的药物组合与在 T 细胞中更有效的药物组合不同。我们的研究表明，HIV 可以在 RTE 细胞中建立潜伏，并且目前的 LRA 仅在诱导 HIV-1 再激活方面略有效果。这表明，为了评估 LRA 作为一种杀菌治愈策略的适用性，可能需要进一步研究非 T 细胞中 LRA 的动态。抗逆转录病毒疗法 (ART) 已大大降低了与 HIV 相关的发病率和死亡率。尽管取得了这一成功，但仍存在一些挑战，包括在没有 ART 的情况下能够重新激活的多种临床潜伏病毒库的长期持续存在。随着人们努力实现 HIV 根除或功能性治愈，有必要进一步了解 HIV-1 复制、潜伏建立和体内携带病毒的储库中再激活的动态。HIV-1 可以感染肾上皮细胞，并且这些细胞中病毒基因的表达有助于未经治疗的个体中 HIV 相关肾病 (HIVAN) 的发展。我们工作的意义在于开发肾上皮细胞中 HIV-1 潜伏的第一个模型。该模型增强了我们对 HIV-1 在肾脏中的潜伏和持续存在的理解，并且可以用于筛选候选潜伏逆转剂。
