Department of Medicine, University of California San Francisco (UCSF).
Department of Medicine, San Francisco VA Medical Center, San Francisco, California, USA.
AIDS. 2020 Nov 15;34(14):2013-2024. doi: 10.1097/QAD.0000000000002684.
While latently HIV-infected cells have been described in the blood, it is unclear whether a similar inducible reservoir exists in the gut, where most HIV-infected cells reside. Tissue-specific environments may contribute to differences in the mechanisms that govern latent HIV infection and amenability to reactivation. We sought to determine whether HIV-infected cells from the blood and gut differ in their responses to T-cell activation and mechanistically distinct latency reversing agents (LRAs).
Cross sectional study using samples from HIV-infected individuals (n = 11).
Matched peripheral blood mononuclear cells (PBMC) and dissociated total cells from rectum ± ileum were treated ex vivo for 24 h with anti-CD3/CD28 or LRAs in the presence of antiretrovirals. HIV DNA and 'read-through', initiated, 5' elongated, completed, and multiply-spliced HIV transcripts were quantified using droplet digital PCR.
T-cell activation increased levels of all HIV transcripts in PBMC and gut cells, and was the only treatment that increased multiply-spliced HIV RNA. Disulfiram increased initiated HIV transcripts in PBMC but not gut cells, while ingenol mebutate increased HIV transcription more in gut cells. Romidepsin increased HIV transcription in PBMC and gut cells, but the increase in transcription initiation was greater in PBMC.
The gut harbors HIV-infected cells in a latent-like state that can be reversed by T-cell activation involving CD3/CD28 signaling. Histone deacetylation and protein kinase B may contribute less to HIV transcriptional initiation in the gut, whereas protein kinase C may contribute more. New LRAs or combinations are needed to induce multiply-spliced HIV and should be tested on both blood and gut.
虽然潜伏的 HIV 感染细胞已在血液中被描述，但尚不清楚是否存在类似的诱导性储库存在于肠道，大多数 HIV 感染细胞存在于肠道中。组织特异性环境可能导致控制潜伏 HIV 感染和易感性的机制存在差异。我们试图确定血液和肠道中的 HIV 感染细胞在对 T 细胞激活的反应以及在机制上不同的潜伏逆转剂 (LRA) 方面是否存在差异。
使用 HIV 感染者的样本进行横断面研究（n=11）。
将外周血单核细胞 (PBMC) 和直肠-回肠分离的总细胞在抗 CD3/CD28 或 LRA 的存在下离体处理 24 小时，同时使用抗逆转录病毒药物。使用液滴数字 PCR 定量 HIV DNA 和“通读”、起始、5'延伸、完成和多剪接 HIV 转录本。
T 细胞激活增加了 PBMC 和肠道细胞中所有 HIV 转录本的水平，并且是唯一增加多剪接 HIV RNA 的治疗方法。双硫仑增加了 PBMC 中的起始 HIV 转录本，但未增加肠道细胞中的 HIV 转录本，而 ingenol mebutate 增加了肠道细胞中的 HIV 转录更多。罗米地辛增加了 PBMC 和肠道细胞中的 HIV 转录，但 PBMC 中的转录起始增加更大。
肠道中存在潜伏样状态的 HIV 感染细胞，可以通过涉及 CD3/CD28 信号的 T 细胞激活来逆转。组蛋白去乙酰化和蛋白激酶 B 可能对肠道中 HIV 转录起始的贡献较小，而蛋白激酶 C 可能贡献更大。需要新的 LRA 或组合来诱导多剪接 HIV，并应在血液和肠道上进行测试。
