Zika Virus Persistently Infects and Is Basolaterally Released from Primary Human Brain Microvascular Endothelial Cells.
Zika Virus Persistently Infects and Is Basolaterally Released from Primary Human Brain Microvascular Endothelial Cells.
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DOI：
10.1128/mbio.00952-17
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发表时间：
2017-07-11
期刊：
影响因子：
6.4
通讯作者：
Mackow ER
中科院分区：
文献类型：
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作者：
Mladinich MC;Schwedes J;Mackow ER
寨卡病毒（ZIKV）是一种由蚊子传播的黄病毒，已成为美洲脑炎和胎儿小脑畸形的原因。ZIKV独特地在人类体液中持续存在长达6个月，通过性传播，并穿过胎盘和血脑屏障（BBB）损害神经元。支持持续ZIKV复制的细胞和ZIKV建立持续性的机制仍然是谜，但对ZIKV进入受保护的神经元区室至关重要。毛细血管的内皮细胞（EC）衬里通常限制经胎盘传递并形成BBB，其选择性地限制血液成分接近神经元。我们发现，ZIKV（毒株PRVABC 59）持续感染原代人脑微血管EC（hBMEC）并在hBMEC传代后在hBMEC中连续复制>9天，而没有细胞病理学。ZIKV不透化hBMEC，但从极化的hBMEC基底外侧释放，表明ZIKV穿过BBB的直接机制。ZIKV感染的hBMEC对α干扰素（IFN-α）具有快速抗性，并且瞬时诱导，但不能分泌IFN-β和IFN-λ。全局转录组分析确定ZIKV在感染后1至9天组成性诱导IFN调节因子7（IRF 7）、IRF 9和IFN刺激基因（ISG），尽管在hBMEC中持续复制。ZIKV组成型诱导ISG 15、HERC 5和USP 18，其与丙型肝炎病毒（HCV）持久性和IFN调节、趋化因子CCL 5以及细胞存活因子相关，趋化因子CCL 5与免疫发病机制相关。我们的研究结果揭示了hBMEC作为ZIKV持续复制的储存库，提示了ZIKV穿过hBMEC进入神经元区室的途径，并定义了ZIKV持续性的新机制，其可以被靶向以限制ZIKV传播。ZIKV在患者中持续存在，穿过胎盘和神经元屏障，损害神经元，并引起胎儿小脑畸形。我们发现ZIKV持续感染脑内皮细胞，这些细胞通常保护神经元免受病毒暴露。hBMEC不被ZIKV感染损伤，并且类似于持续性HCV感染，ZIKV组成型诱导和逃避抗病毒ISG和IFN应答以在hBMEC中连续复制。因此，hBMEC为全身性ZIKV传播提供了保护性小生境，并提供了位于正常保护性血脑屏障中的病毒储库。与ZIKV扩散到神经元区室中一致，ZIKV从hBMEC基底外侧释放。我们的研究结果定义了有助于持续ZIKV感染的hBMEC应答和从hBMEC清除ZIKV感染的潜在靶标。这些结果进一步表明了额外的ZIKV感染的EC促进病毒在受保护的胎盘、视网膜和睾丸隔室中传播和持续存在的作用。
Zika virus (ZIKV) is a mosquito-borne Flavivirus that has emerged as the cause of encephalitis and fetal microencephaly in the Americas. ZIKV uniquely persists in human bodily fluids for up to 6 months, is sexually transmitted, and traverses the placenta and the blood-brain barrier (BBB) to damage neurons. Cells that support persistent ZIKV replication and mechanisms by which ZIKV establishes persistence remain enigmatic but central to ZIKV entry into protected neuronal compartments. The endothelial cell (EC) lining of capillaries normally constrains transplacental transmission and forms the BBB, which selectively restricts access of blood constituents to neurons. We found that ZIKV (strain PRVABC59) persistently infects and continuously replicates in primary human brain microvascular ECs (hBMECs), without cytopathology, for >9 days and following hBMEC passage. ZIKV did not permeabilize hBMECs but was released basolaterally from polarized hBMECs, suggesting a direct mechanism for ZIKV to cross the BBB. ZIKV-infected hBMECs were rapidly resistant to alpha interferon (IFN-α) and transiently induced, but failed to secrete, IFN-β and IFN-λ. Global transcriptome analysis determined that ZIKV constitutively induced IFN regulatory factor 7 (IRF7), IRF9, and IFN-stimulated genes (ISGs) 1 to 9 days postinfection, despite persistently replicating in hBMECs. ZIKV constitutively induced ISG15, HERC5, and USP18, which are linked to hepatitis C virus (HCV) persistence and IFN regulation, chemokine CCL5, which is associated with immunopathogenesis, as well as cell survival factors. Our results reveal that hBMECs act as a reservoir of persistent ZIKV replication, suggest routes for ZIKV to cross hBMECs into neuronal compartments, and define novel mechanisms of ZIKV persistence that can be targeted to restrict ZIKV spread. ZIKV persists in patients, crossing placental and neuronal barriers, damaging neurons, and causing fetal microencephaly. We found that ZIKV persistently infects brain endothelial cells that normally protect neurons from viral exposure. hBMECs are not damaged by ZIKV infection and, analogous to persistent HCV infection, ZIKV constitutively induces and evades antiviral ISG and IFN responses to continuously replicate in hBMECs. As a result, hBMECs provide a protective niche for systemic ZIKV spread and a viral reservoir localized in the normally protective blood-brain barrier. Consistent with the spread of ZIKV into neuronal compartments, ZIKV was released basolaterally from hBMECs. Our findings define hBMEC responses that contribute to persistent ZIKV infection and potential targets for clearing ZIKV infections from hBMECs. These results further suggest roles for additional ZIKV-infected ECs to facilitate viral spread and persistence in the protected placental, retinal, and testicular compartments.