Tetherin can restrict cell-free and cell-cell transmission of HIV from primary macrophages to T cells.
Tetherin can restrict cell-free and cell-cell transmission of HIV from primary macrophages to T cells.
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DOI：
10.1371/journal.ppat.1004189
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发表时间：
2014-07
期刊：
影响因子：
6.7
通讯作者：
Marsh M
中科院分区：
文献类型：
--
作者：
Giese S;Marsh M
BST-2/Tetherin通过将新形成的病毒颗粒束缚在受感染细胞的质膜上来抑制HIV的释放。虽然Tetherin介导的限制机制越来越清楚，但这种限制在HIV天然靶细胞中的生物学相关性尚不清楚。此外，Tetherin是否对HIV通过细胞间接触的直接细胞间传播施加任何限制仍然存在争议。在这里，我们分析了限制内源性Tetherin强加给HIV传播的主要人类巨噬细胞，HIV在体内的主要目标之一。我们发现巨噬细胞中Tetherin的mRNA和蛋白水平与来自相同供体的T细胞中的Tetherin的mRNA和蛋白水平相当，并且I型干扰素高度上调。改进的免疫细胞化学方案使我们能够证明Tetherin定位于细胞表面，trans-Golgi网络和巨噬细胞HIV组装室。系链蛋白将出芽的病毒体保留在组装区室中，从而阻碍HIV的释放和无细胞传播，但它本身并不是维持这些区室所必需的。值得注意的是，使用一种新的测定来量化细胞间的扩散，我们表明Tetherin促进病毒簇从巨噬细胞转移到T细胞，从而限制了双嗜性HIV-1的直接传播。动力学分析提供了支持的概念，这种直接巨噬细胞-T细胞传播介导的，至少在一定程度上，由所谓的病毒突触。最后，我们证明了病毒Vpu蛋白有效地下调细胞表面和Tetherin的总体水平，从而消除了巨噬细胞中的这种HIV限制。总之，我们的研究表明，Tetherin是迄今为止发现的最有效的HIV限制因子之一，可以抑制病毒从原代巨噬细胞传播，无论传播方式如何。Tetherin是一种细胞蛋白质，通过将新生颗粒物理“束缚”在受感染细胞的质膜上来抑制（或限制）包括艾滋病毒在内的多种包膜病毒。CD 4 + T细胞和巨噬细胞是HIV在体内的主要靶标，并且这两种细胞类型都表达Tetherin。虽然模型细胞系和T细胞中的Tetherin介导的限制机制越来越清楚，但巨噬细胞的实验数据很少，并且与其他细胞类型中的观察结果部分矛盾。在这里，我们调查的敏感性Tetherin表达干扰素，和亚细胞定位的限制因子在原代人类巨噬细胞。我们发现Tetherin通过在巨噬细胞HIV组装室中保留新生颗粒来抑制HIV释放，并且还可以限制HIV在巨噬细胞和T细胞之间的细胞间接触中的传播。最后，我们证明了HIV蛋白Vpu有效地抵消了巨噬细胞中的Tetherin，从而确保了病毒的繁殖。我们的研究结果以及其他已发表的数据表明，Tetherin可以有效地抑制HIV两种主要靶细胞类型中的病毒复制，无论其传播方式如何。这些数据支持了这样一种观点，即Tetherin的有效对抗是艾滋病毒全球传播的关键因素。
Bst-2/Tetherin inhibits the release of HIV by tethering newly formed virus particles to the plasma membrane of infected cells. Although the mechanisms of Tetherin-mediated restriction are increasingly well understood, the biological relevance of this restriction in the natural target cells of HIV is unclear. Moreover, whether Tetherin exerts any restriction on the direct cell-cell spread of HIV across intercellular contacts remains controversial. Here we analyse the restriction endogenous Tetherin imposes on HIV transmission from primary human macrophages, one of the main targets of HIV in vivo. We find that the mRNA and protein levels of Tetherin in macrophages are comparable to those in T cells from the same donors, and are highly upregulated by type I interferons. Improved immunocytochemistry protocols enable us to demonstrate that Tetherin localises to the cell surface, the trans-Golgi network, and the macrophage HIV assembly compartments. Tetherin retains budded virions in the assembly compartments, thereby impeding the release and cell-free spread of HIV, but it is not required for the maintenance of these compartments per se. Notably, using a novel assay to quantify cell-cell spread, we show that Tetherin promotes the transfer of virus clusters from macrophages to T cells and thereby restricts the direct transmission of a dual-tropic HIV-1. Kinetic analyses provide support for the notion that this direct macrophage-T cell spread is mediated, at least in part, by so-called virological synapses. Finally, we demonstrate that the viral Vpu protein efficiently downregulates the cell surface and overall levels of Tetherin, and thereby abrogates this HIV restriction in macrophages. Together, our study shows that Tetherin, one of the most potent HIV restriction factors identified to date, can inhibit virus spread from primary macrophages, regardless of the mode of transmission. Tetherin is a cellular protein that inhibits (or restricts) a broad range of enveloped viruses, including HIV, by physically “tethering” nascent particles to the plasma membrane of infected cells. CD4+ T cells and macrophages are the main targets of HIV in vivo, and both cell types express Tetherin. Although the mechanisms of Tetherin-mediated restriction in model cell lines and T cells are increasingly well understood, experimental data from macrophages are sparse, and partially contradict observations made in other cell types. Here we investigate the sensitivity of Tetherin expression to interferon, and the subcellular localisation of the restriction factor in primary human macrophages. We find that Tetherin inhibits HIV release by retaining nascent particles in macrophage HIV assembly compartments, and can also restrict the transmission of HIV across intercellular contacts between macrophages and T cells. Finally, we demonstrate that the HIV protein Vpu efficiently counteracts Tetherin in macrophages, and thereby ensures viral propagation. Our results, together with other published data, show that Tetherin can efficiently inhibit viral replication in both major target cell types of HIV, regardless of the mode of transmission. These data support the view that efficient counteraction of Tetherin was a crucial factor for the global spread of HIV.
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影响因子：
6.7
作者：
Casartelli N;Sourisseau M;Feldmann J;Guivel-Benhassine F;Mallet A;Marcelin AG;Guatelli J;Schwartz O
通讯作者：
Schwartz O
DOI：
10.1083/jcb.200609050
发表时间：
2007-04-23
期刊：
The Journal of cell biology
影响因子：
--
作者：
Deneka M;Pelchen-Matthews A;Byland R;Ruiz-Mateos E;Marsh M
通讯作者：
Marsh M
影响因子：
30.3
作者：
Jouve, Mabel;Sol-Foulon, Nathalie;Benaroch, Philippe
通讯作者：
Benaroch, Philippe
影响因子：
6.7
作者：
Janvier K;Pelchen-Matthews A;Renaud JB;Caillet M;Marsh M;Berlioz-Torrent C
通讯作者：
Berlioz-Torrent C
影响因子：
3.3
作者：
Kuhl BD;Sloan RD;Donahue DA;Bar-Magen T;Liang C;Wainberg MA
通讯作者：
Wainberg MA