Single-Particle Tracking of Human Immunodeficiency Virus Type 1 Productive Entry into Human Primary Macrophages
Single-Particle Tracking of Human Immunodeficiency Virus Type 1 Productive Entry into Human Primary Macrophages
复制标题
单粒子追踪人类免疫缺陷病毒 1 型有效进入人类原代巨噬细胞
DOI：
10.1021/acsnano.7b00275
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发表时间：
2017-04-01
期刊：
影响因子：
17.1
通讯作者：
Cui, Zongqiang
中科院分区：
文献类型：
--
作者：
Li, Qin;Li, Wei;Cui, Zongqiang
巨噬细胞是人类免疫缺陷病毒（HIV-1）的主要靶点之一，但病毒进入这些细胞的途径仍然知之甚少。非侵入性病毒标记和单病毒追踪是研究病毒入侵的有效工具。在这里，我们构建了一个量子点（QD）封装的感染性HIV-1颗粒，以跟踪病毒进入活的人原代巨噬细胞在单颗粒水平。通过在病毒组装过程中掺入病毒辅助蛋白Vpr缀合的QD，将QD包封在HIV-1病毒体中。利用包裹量子点的HIV-1颗粒，我们真实的监测了病毒感染的早期阶段，并观察到，在感染过程中，HIV-1以网格蛋白介导的方式被内吞;颗粒易位到Rab 5A阳性内体中，核心通过病毒囊泡介导的内体融合释放到细胞质中。药物抑制试验证实，内体融合有助于HIV-1在原代巨噬细胞中的生产性感染。此外，我们观察到，动态肌动蛋白细胞骨架是至关重要的HIV-1进入和细胞内迁移在原代巨噬细胞。HIV-1动力学和感染可以被多种不同的肌动蛋白抑制剂阻断。我们的研究揭示了巨噬细胞中的生产性进入途径，该途径需要内体功能和肌动蛋白动力学，这可能有助于开发抑制剂以阻断巨噬细胞中的HIV进入。
Macrophages are one of the major targets of human immunodeficiency virus (HIV-1), but the viral entry pathway remains poorly understood in these cells. Noninvasive virus labeling and single-virus tracking are effective tools for studying virus entry. Here, we constructed a quantum dot (QD)-encapsulated infectious HIV-1 particle to track viral entry at a single-particle level in live human primary macrophages. QDs were encapsulated in HIV-1 virions by incorporating viral accessory protein Vpr-conjugated QDs during virus assembly. With the HIV-1 particles encapsulating QDs, we monitored the early phase of viral infection in real time and observed that, during infection, HIV-1 was endocytosed in a clathrin-mediated manner; the particles were translocated into Rab5A-positive endosomes, and the core was released into the cytoplasm by viral envelope-mediated endosomal fusion. Drug inhibition assays verified that endosome fusion contributes to HIV-1 productive infection in primary macrophages. Additionally, we observed that a dynamic actin cytoskeleton is critical for HIV-1 entry and intracellular migration in primary macrophages. HIV-1 dynamics and infection could be blocked by multiple different actin inhibitors. Our study revealed a productive entry pathway in macrophages that requires both endosomal function and actin dynamics, which may assist in the development of inhibitors to block the HIV entry in macrophages.