Exosomes from Human Immunodeficiency Virus Type 1 (HIV-1)-Infected Cells License Quiescent CD4+ T Lymphocytes To Replicate HIV-1 through a Nef- and ADAM17-Dependent Mechanism
Exosomes from Human Immunodeficiency Virus Type 1 (HIV-1)-Infected Cells License Quiescent CD4+ T Lymphocytes To Replicate HIV-1 through a Nef- and ADAM17-Dependent Mechanism
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DOI：
10.1128/jvi.01712-14
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发表时间：
2014-10-01
影响因子：
5.4
通讯作者：
Federico, Maurizio
中科院分区：
文献类型：
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作者：
Arenaccio, Claudia;Chiozzini, Chiara;Federico, Maurizio
静息的CD 4（+）T淋巴细胞抵抗人类免疫缺陷病毒（HIV）感染。在这里，我们提供的证据表明，来自HIV-1感染细胞的外泌体使静息的人原代CD 4（+）T淋巴细胞允许HIV-1复制。这些结果是在存在外泌体释放抑制剂的情况下，用HIV-1感染细胞与静止的CD 4（+）T淋巴细胞的transwell共培养物获得的，并使用从HIV-1感染的原代CD 4（+）T淋巴细胞的上清液中纯化的外泌体证实。我们发现，HIV-1 Nef在产生外泌体的细胞中的表达对于细胞活化以及HIV-1在靶CD 4（+）T淋巴细胞中的复制是必要的和充分的。我们还确定了一个对我们观察到的效应很重要的Nef结构域，即，（EEEE 65）-E-62酸性簇结构域。此外，我们观察到ADAM 17，即，一种成熟形式的去整合素和金属蛋白酶，转化前肿瘤坏死因子α（TNF-α），与来自HIV-1感染细胞的外来体相关，在静止的CD 4（+）T淋巴细胞中HIV-1复制中起关键作用。用ADAM 17抑制剂治疗可消除静息CD 4（+）T淋巴细胞的活化和HIV-1复制。TNF-α是ADAM 17的下游效应子，因为用抗TNF-α抗体处理静息淋巴细胞阻断了HIV-1的复制。这里呈现的数据与Nef通过外泌体诱导细胞间通讯以激活旁观者静止的CD 4（+）T淋巴细胞从而刺激病毒传播的模型一致。重要性总体而言，我们的研究结果支持HIV进化为篡夺基于外泌体的细胞间通讯网络以促进其在感染宿主中传播的观点。
Resting CD4(+) T lymphocytes resist human immunodeficiency virus (HIV) infection. Here, we provide evidence that exosomes from HIV-1-infected cells render resting human primary CD4(+) T lymphocytes permissive to HIV-1 replication. These results were obtained with transwell cocultures of HIV-1-infected cells with quiescent CD4(+) T lymphocytes in the presence of inhibitors of exosome release and were confirmed using exosomes purified from supernatants of HIV-1-infected primary CD4(+) T lymphocytes. We found that the expression of HIV-1 Nef in exosome-producing cells is both necessary and sufficient for cell activation as well as HIV-1 replication in target CD4(+) T lymphocytes. We also identified a Nef domain important for the effects we observed, i.e., the (EEEE65)-E-62 acidic cluster domain. In addition, we observed that ADAM17, i.e., a disintegrin and metalloprotease converting pro-tumor necrosis factor alpha (TNF-alpha) in its mature form, associates with exosomes from HIV-1-infected cells, and plays a key role in the HIV-1 replication in quiescent CD4(+) T lymphocytes. Treatment with an inhibitor of ADAM17 abolished both activation and HIV-1 replication in resting CD4(+) T lymphocytes. TNF-alpha is the downstream effector of ADAM17 since the treatment of resting lymphocytes with anti-TNF-alpha antibodies blocked the HIV-1 replication. The data presented here are consistent with a model where Nef induces intercellular communication through exosomes to activate bystander quiescent CD4(+) T lymphocytes, thus stimulating viral spread.IMPORTANCEOverall, our findings support the idea that HIV evolved to usurp the exosome-based intercellular communication network to favor its spread in infected hosts.