Sarabia Indra, Huang Szu-Han, Ward Adam R, Jones R Brad, Bosque Alberto
Department of Microbiology, Immunology and Tropical Medicine, George Washington University, Washington, USA.
Infectious Disease Division, Weill Cornell Medical College, New York, USA.
J Virol. 2021 Mar 10;95(7). doi: 10.1128/JVI.01297-20. Epub 2021 Jan 13.
The establishment of HIV-1 latency has hindered an HIV-1 cure. "Shock and Kill" strategies to target this reservoir aim to induce the latent provirus with latency reversing agents (LRAs). However, recent studies have shown that the majority of the intact HIV-1 viral reservoir found in ART-suppressed HIV infected individuals is not inducible. We sought to understand whether this non-inducible reservoir is established, and thus able to be studied, in an primary T model of latency. Furthermore, we wanted to expand this model system to include R5-tropic and non-B subtype viruses. To that end, we generated our T model of latency with an R5 subtype B virus, AD8 and an R5 subtype C virus, MJ4. Our results demonstrate that both intact and defective proviruses are generated in this model. Less than 50% of intact proviruses are inducible regardless of viral strain in the context of maximal stimulation through the TCR or with different clinically relevant LRAs including the HDAC inhibitors SAHA and MS-275, the PKC agonist Ingenol 3,20-dibenzoate or the SMAC mimetic AZD-5582. Our findings suggest that current LRA strategies are insufficient to effectively reactivate intact latent HIV-1 proviruses in primary CD4 T cells and that the mechanisms involved in the generation of the non-inducible HIV-1 reservoir can be studied using this primary model. HIV-1 establishes a latent reservoir that persists under antiretroviral therapy. Antiretroviral therapy is able to stop the spread of the virus and the progression of the disease but does not target this latent reservoir. If antiretroviral therapy is stopped, the virus is able to resume replication and the disease progresses. Recently, it has been demonstrated that most of the latent reservoir capable of generating replication competent virus cannot be induced in the laboratory setting. However, the mechanisms that influence the generation of this intact and non-inducible latent reservoir are still under investigation. Here we demonstrate the generation of defective, intact and intact non-inducible latent HIV-1 in a T model of latency using different HIV-1 strains. Thus, the mechanisms which control inducibility can be studied using this primary cell model of latency, which may accelerate our understanding of the latent reservoir and the development of curative strategies.
HIV-1潜伏状态的建立阻碍了对HIV-1的治愈。针对该病毒储存库的“激活并清除”策略旨在用潜伏逆转剂（LRA）诱导潜伏的前病毒。然而，最近的研究表明，在接受抗逆转录病毒治疗（ART）的HIV感染者中发现的大多数完整HIV-1病毒储存库是不可诱导的。我们试图了解这种不可诱导的储存库是否在原发性T细胞潜伏模型中得以建立，从而能够进行研究。此外，我们希望扩展这个模型系统，使其包括R5嗜性和非B亚型病毒。为此，我们用R5 B亚型病毒AD8和R5 C亚型病毒MJ4构建了我们的T细胞潜伏模型。我们的结果表明，在这个模型中会产生完整和缺陷的前病毒。在通过TCR进行最大刺激的情况下，或者使用不同的具有临床相关性的LRA（包括组蛋白去乙酰化酶抑制剂SAHA和MS-275、蛋白激酶C激动剂Ingenol 3,20-二苯甲酸酯或SMAC模拟物AZD-5582）时，无论病毒株如何，不到50%的完整前病毒是可诱导的。我们的研究结果表明，目前的LRA策略不足以有效地重新激活原发性CD⁺T细胞中完整的潜伏HIV-1前病毒，并且可以使用这个原发性细胞模型来研究不可诱导的HIV-1储存库产生所涉及的机制。HIV-1建立了一个在抗逆转录病毒治疗下持续存在的潜伏储存库。抗逆转录病毒治疗能够阻止病毒传播和疾病进展，但并不针对这个潜伏储存库。如果停止抗逆转录病毒治疗，病毒能够恢复复制，疾病会继续发展。最近，已经证明在实验室环境中，大多数能够产生具有复制能力病毒的潜伏储存库是不可诱导的。然而，影响这个完整且不可诱导的潜伏储存库产生的机制仍在研究中。在这里，我们展示了在T细胞潜伏模型中使用不同的HIV-1毒株产生缺陷型、完整型和完整不可诱导型潜伏HIV-1的情况。因此，可以使用这个原发性细胞潜伏模型来研究控制诱导性的机制，这可能会加速我们对潜伏储存库的理解以及治愈策略的开发。
