Latency reversal agents affect differently the latent reservoir present in distinct CD4+ T subpopulations
Latency reversal agents affect differently the latent reservoir present in distinct CD4+ T subpopulations
复制标题
DOI：
10.1371/journal.ppat.1007991
复制
发表时间：
2019-08-01
期刊：
影响因子：
6.7
通讯作者：
Buzon, Maria J.
中科院分区：
文献类型：
--
作者：
Grau-Exposito, Judith;Luque-Ballesteros, Laura;Buzon, Maria J.
潜伏期逆转剂 (LRAs) 已被证明可以在体内诱导 HIV-1 转录，但无法有效减少接受抗逆转录病毒治疗的患者中潜伏病毒库的大小。 LRA 扰乱不同细胞亚群中病毒库的能力目前尚不清楚。在这里，我们使用新的 RNA FISH/flow 离体病毒再激活测定，对不同的 LRA 家族及其组合在不同 CD4(+) T 细胞亚群中的病毒再激活能力进行了全面评估。我们观察到，病毒重新激活后，整个 HIV 储存库的中位数为 16.28% 诱导 HIV-1 转录，但这些 HIV-1 RNA(+) 细胞中只有 10.10% 产生病毒蛋白 p24。此外，没有一个 LRA 强大到足以重新激活所有 CD4 + T 细胞亚群中的 HIV-1 转录。例如，Romidepsin 和 Ingenol 的组合被认为是增加大多数（但不是全部）CD4（+）T 细胞亚群中 HIV-1 RNA（+）细胞比例的最佳药物组合。重要的是，记忆干细胞被认为对 HIV-1 重新激活具有高度抵抗力，只有帕比司他和 Bryostatin-1 的组合才能显着增加该亚群内转录 HIV 的细胞数量。总体而言，我们的结果验证了使用RNA FISH/流式技术来评估不同CD4（+）T细胞亚群中LRAs的效力，显示了包含潜伏HIV储存库的细胞之间的内在差异，并强调了用目前可用的药物显着影响潜伏感染的难度。因此，我们的结果对于合理设计旨在逆转不同细胞储存库的 HIV 潜伏期的疗法具有重要意义。
Latency reversal agents (LRAs) have proven to induce HIV-1 transcription in vivo but are ineffective at decreasing the size of the latent reservoir in antiretroviral treated patients. The capacity of the LRAs to perturb the viral reservoir present in distinct subpopulations of cells is currently unknown. Here, using a new RNA FISH/flow ex vivo viral reactivation assay, we performed a comprehensive assessment of the viral reactivation capacity of different families of LRAs, and their combinations, in different CD4(+) T cell subsets. We observed that a median of 16.28% of the whole HIV-reservoir induced HIV-1 transcripts after viral reactivation, but only 10.10% of these HIV-1 RNA(+) cells produced the viral protein p24. Moreover, none of the LRAs were powerful enough to reactivate HIV-1 transcription in all CD4 + T cell subpopulations. For instance, the combination of Romidepsin and Ingenol was identified as the best combination of drugs at increasing the proportion of HIV-1 RNA(+) cells, in most, but not all, CD4(+) T cell subsets. Importantly, memory stem cells were identified as highly resistant to HIV-1 reactivation, and only the combination of Panobinostat and Bryostatin-1 significantly increased the number of cells transcribing HIV within this subset. Overall, our results validate the use of the RNA FISH/flow technique to assess the potency of LRAs among different CD4(+) T cell subsets, manifest the intrinsic differences between cells that encompass the latent HIV reservoir, and highlight the difficulty to significantly impact the latent infection with the currently available drugs. Thus, our results have important implications for the rational design of therapies aimed at reversing HIV latency from diverse cellular reservoirs.