Department of Infectious Diseases, The University of Melbourne at The Peter Doherty Institute for Infection and Immunity, Melbourne, Australia.
Department of Infectious Diseases, The University of Melbourne at The Peter Doherty Institute for Infection and Immunity, Melbourne, Australia; Department of Microbiology and Immunology, The University of Melbourne at the Peter Doherty Institute for Infection and Immunity, Melbourne, Australia.
EBioMedicine. 2021 Mar;65:103241. doi: 10.1016/j.ebiom.2021.103241. Epub 2021 Feb 26.
One strategy being pursued to clear latently infected cells that persist in people living with HIV (PLWH) on antiretroviral therapy (ART) is to activate latent HIV infection with a latency reversing agent (LRA). Surrogate markers that accurately measure virus production following an LRA are needed.
We quantified cell-associated unspliced (US), multiply spliced (MS) and supernatant (SN) HIV RNA by qPCR from total and resting CD4+ T cells isolated from seven PLWH on ART before and after treatment ex vivo with different LRAs, including histone deacetylase inhibitors (HDACi). MS and plasma HIV RNA were also quantified from PLWH on ART (n-11) who received the HDACi panobinostat.
In total and resting CD4+ T cells from PLWH on ART, detection of US RNA was common while detection of MS RNA was infrequent. Primers used to detect MS RNA, in contrast to US RNA, bound sites of the viral genome that are commonly mutated or deleted in PLWH on ART. Following ex vivo stimulation with LRAs, we identified a strong correlation between the fold change increase in SN and MS RNA, but not the fold change increase in SN and US RNA. In PLWH on ART who received panobinostat, MS RNA was significantly higher in samples with detectable compared to non0detectable plasma HIV RNA.
Following administration of an LRA, quantification of MS RNA is more likely to reflect an increase in virion production and is therefore a better indicator of meaningful latency reversal.
NHMRC, NIH DARE collaboratory.
目前，人们正在探索一种策略来清除接受抗逆转录病毒疗法（ART）的艾滋病毒感染者（PLWH）体内持续存在的潜伏感染细胞，该策略是使用潜伏逆转剂（LRA）激活潜伏的 HIV 感染。目前需要一种能够准确测量 LRA 后病毒产生的替代标志物。
我们通过 qPCR 定量检测了来自七位接受 ART 治疗的 PLWH 总 CD4+T 细胞和静息 CD4+T 细胞中细胞相关未剪接（US）、多剪接（MS）和上清液（SN）HIV RNA，这些细胞在接受不同 LRA（包括组蛋白去乙酰化酶抑制剂（HDACi））的体外治疗前后进行了分离。我们还从接受 HDACi 帕比司他治疗的接受 ART 治疗的 PLWH（n=11）中定量检测了 MS 和血浆 HIV RNA。
在接受 ART 治疗的 PLWH 的总 CD4+T 细胞和静息 CD4+T 细胞中，US RNA 的检测很常见，而 MS RNA 的检测则很少见。与 US RNA 不同，用于检测 MS RNA 的引物结合了在接受 ART 治疗的 PLWH 中常见突变或缺失的病毒基因组位点。在体外接受 LRA 刺激后，我们发现 SN 和 MS RNA 的倍数变化增加之间存在很强的相关性，但 SN 和 US RNA 的倍数变化增加之间没有相关性。在接受帕比司他治疗的接受 ART 治疗的 PLWH 中，与不可检测到的血浆 HIV RNA 相比，可检测到的血浆 HIV RNA 样本中的 MS RNA 明显更高。
在给予 LRA 后，MS RNA 的定量更有可能反映出病毒产生的增加，因此是潜伏逆转的更有意义的指标。
澳大利亚国家健康与医学研究委员会，美国国立卫生研究院 DARE 合作研究中心。
