Prolonged persistence of a novel replication-defective HIV-1 variant in plasma of a patient on suppressive therapy.
Prolonged persistence of a novel replication-defective HIV-1 variant in plasma of a patient on suppressive therapy.
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DOI：
10.1186/s12985-016-0617-0
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发表时间：
2016-09-21
期刊：
影响因子：
4.8
通讯作者：
Sahu GK
中科院分区：
文献类型：
--
作者：
Rassler S;Ramirez R;Khoury N;Skowron G;Sahu GK
在大多数接受抑制性抗逆转录病毒治疗（ART）的患者的血浆中，无细胞残留HIV-1病毒粒子（RVs）持续低于20-50 vRNA拷贝/ml。在治疗过程中，rv是如何在体内产生的尚不完全清楚。在这项研究中，我们试图在体外对接受art治疗的患者的这些病毒进行表征，以便深入了解它们在体内产生的机制。我们利用患者残留的血浆vRNA，通过重叠rt -巢式PCR方法，以DNA形式重建了几乎整个rv基因组，然后对克隆基因组进行了测序分析，并在体外测试了它们的生物活性。我们发现重建的RVs分子克隆缺乏抗逆转录病毒耐药突变，以及G-to-A超突变。当将vDNA克隆转染到TZM-bl细胞中时，将HIV-p24以极低的水平释放到培养基中。发现这种低水平的病毒产生是由于在相应vrna的保守的5 ' -主剪接供体（MSD）基序中存在独特的突变（GU-to-GC）。我们发现这个点突变本身的掺入会导致标准HIV毒株（JRCSF）在体外复制的缺陷。然而，在39个月的治疗期间，在患者血浆的7个时间点中有5个时断时续地检测到这种新型病毒变体。这是首次在HIV基因组RNA保守的5 ' -MSD基序中发现自然点突变（GU-to-GC）。在患者血浆和其他病毒群中间歇性但长时间地检测到这种复制缺陷HIV变体，强烈表明这种变体在治疗期间从体内存在的高度稳定的生产感染细胞中释放出来。这一观察结果的潜在含义是，在抑制治疗期间消除这种导致残留病毒血症的生产性感染细胞可能是实现治愈艾滋病毒的重要的第一步。本文的在线版本（doi:10.1186/s12985-016-0617-0）包含补充材料，仅供授权用户使用。
Cell-free residual HIV-1 virions (RVs) persist in plasma below 20–50 vRNA copies/ml in most patients on suppressive antiretroviral therapy (ART). How RVs are produced in the body during therapy is not fully clear. In this study, we have attempted to characterize these viruses of an ART-treated patient in vitro in order to gain insights into the mechanism of their production in vivo. We have reconstructed almost the entire genomes of RVs as DNA forms using the patient’s residual plasma vRNA by an overlapping RT-nested PCR method, and then sequence-analyzed the cloned genomes and tested them for their biological activities in vitro. We found that the reconstructed molecular clones of RVs lacked antiretroviral drug-resistant mutations, as well as G-to-A hypermutations. The vDNA clones, when transfected into TZM-bl cells, released HIV-p24 into the culture media at extremely low levels. This low-level virus production was found to be due to the presence of a unique mutation (GU-to-GC) in the conserved 5′-major splice donor (MSD) motif of the corresponding vRNAs. We found that the incorporation of this point mutation by itself could cause defects in the replication of a standard HIV strain (JRCSF) in vitro. However, this novel viral variant was intermittently detected at 5 of 7 time-points in the patient’s plasma over a period of 39 months during therapy. This is the first identification of a natural point mutation (GU-to-GC) in the conserved 5′-MSD motif of HIV genomic RNA. The intermittent but prolonged detection of this replication-defective HIV variant in the patient′s plasma among other viral populations strongly suggests that this variant is released from highly stable productively infected cells present in vivo during therapy. The potential implication of this observation is that the elimination of such productively infected cells that contribute to residual viremia during suppressive therapy could be an important first step towards achieving a cure for HIV. The online version of this article (doi:10.1186/s12985-016-0617-0) contains supplementary material, which is available to authorized users.
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