Detailed Characterization of Early HIV-1 Replication Dynamics in Primary Human Macrophages.
Detailed Characterization of Early HIV-1 Replication Dynamics in Primary Human Macrophages.
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DOI：
10.3390/v10110620
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发表时间：
2018-11-10
期刊：
影响因子：
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通讯作者：
Kräusslich HG
中科院分区：
文献类型：
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作者：
Bejarano DA;Puertas MC;Börner K;Martinez-Picado J;Müller B;Kräusslich HG
巨噬细胞是人类免疫缺陷病毒1型（HIV-1）的天然靶细胞。与淋巴细胞相比，这些细胞中的病毒复制似乎被延迟了；然而，对早期进入后事件的动力学知之甚少。使用几种HIV-1抑制剂进行的添加时间实验和用液滴数字PCR （ddPCR）检测逆转录酶（RT）产物显示，在一些供者的原代人单核细胞来源的巨噬细胞中，早期复制被延迟，并在感染后达到峰值。通过点击标记新合成的DNA，对逆转录和预整合复合物（RTC/PIC）进行直接成像进一步证实了我们的发现，并显示随着时间的推移，逆转录和预整合复合物会随之向核阶段转移。改变进入途径增强了传染性，但不影响病毒复制动力学。病毒蛋白X （Vpx）的加入增强了多产性感染，加速了逆转录和核进入的完成。我们认为，无菌α基序（SAM）和组氨酸/天冬氨酸（HD）结构域含蛋白1 （SAMHD1）活性降低的脱氧核糖核苷酸三磷酸（dNTP）池是延迟巨噬细胞早期HIV-1复制的主要因素。
Macrophages are natural target cells of human immunodeficiency virus type 1 (HIV-1). Viral replication appears to be delayed in these cells compared to lymphocytes; however, little is known about the kinetics of early post-entry events. Time-of-addition experiments using several HIV-1 inhibitors and the detection of reverse transcriptase (RT) products with droplet digital PCR (ddPCR) revealed that early replication was delayed in primary human monocyte-derived macrophages of several donors and peaked late after infection. Direct imaging of reverse-transcription and pre-integration complexes (RTC/PIC) by click-labeling of newly synthesized DNA further confirmed our findings and showed a concomitant shift to the nuclear stage over time. Altering the entry pathway enhanced infectivity but did not affect kinetics of viral replication. The addition of viral protein X (Vpx) enhanced productive infection and accelerated completion of reverse transcription and nuclear entry. We propose that sterile alpha motif (SAM) and histidine/aspartate (HD) domain-containing protein 1 (SAMHD1) activity lowering deoxyribonucleotide triphosphate (dNTP) pools is the principal factor delaying early HIV-1 replication in macrophages.