A Three-Dimensional Cell Culture System To Model RNA Virus Infections at the Blood-Brain Barrier.
A Three-Dimensional Cell Culture System To Model RNA Virus Infections at the Blood-Brain Barrier.
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DOI：
10.1128/msphere.00206-17
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发表时间：
2017-05
期刊：
影响因子：
4.8
通讯作者：
Coyne CB
中科院分区：
文献类型：
--
作者：
Bramley JC;Drummond CG;Lennemann NJ;Good CA;Kim KS;Coyne CB
嗜神经病毒感染是全球发病率和死亡率的重要来源。血脑屏障（BBB）部分由微血管内皮细胞层组成，其功能是限制病毒进入大脑。缺乏概括人BBB内皮的许多特性的体外模型，特别是关于这些细胞限制大脑病毒感染的独特细胞和免疫机制。在这里，我们开发了一种三维细胞培养模型，该模型概括了BBB微血管的许多形态和功能特性，并将该模型应用于RNA病毒感染的研究。因此，我们描述的模型可用于研究BBB生理学的各个方面，包括病毒可能进入CNS的机制，并可用于开发和筛选抗病毒治疗药物，以限制病毒发病机制中的这一重要步骤。血脑屏障（BBB）包括脑中最重要的保护屏障，并且部分由衬在脑周围的毛细血管上的微血管内皮细胞层组成。在这里，我们描述了一个人血脑屏障微血管内皮细胞的三维（3-D）模型，该模型概括了这些细胞在体内的特性，包括生理相关的转录谱，诱导有效的抗微生物先天免疫信号传导的能力，以及抵抗各种RNA病毒（包括肠道病毒成员）感染的能力。（柯萨奇病毒B、埃可病毒11、肠道病毒71、脊髓灰质炎病毒）和黄病毒（登革热病毒、寨卡病毒[ZIKV]）家族。我们表明，促炎细胞因子肿瘤坏死因子α（TNF-α）对顶端紧密连接的破坏使3-D培养的BBB细胞对ZIKV感染敏感，并且3-D衍生的BBB细胞可用于模拟ZIKV感染的单核细胞穿过内皮屏障进入底层星形胶质细胞的迁移。总之，我们的研究结果表明，在3-D中培养的人BBB微血管内皮细胞可用于模拟RNA病毒进入中枢神经系统（CNS）的机制，这可用于开发和筛选治疗方法以限制这种事件。重要性嗜神经病毒感染是全球发病率和死亡率的重要来源。血脑屏障（BBB）部分由微血管内皮细胞层组成，其功能是限制病毒进入大脑。缺乏概括人BBB内皮的许多特性的体外模型，特别是关于这些细胞限制大脑病毒感染的独特细胞和免疫机制。在这里，我们开发了一种三维细胞培养模型，该模型概括了BBB微血管的许多形态和功能特性，并将该模型应用于RNA病毒感染的研究。因此，我们描述的模型可用于研究BBB生理学的各个方面，包括病毒可能进入CNS的机制，并可用于开发和筛选抗病毒治疗药物，以限制病毒发病机制中的这一重要步骤。
Neurotropic viral infections are significant sources of global morbidity and mortality. The blood-brain barrier (BBB) is composed in part of a layer of microvascular endothelial cells and functions to restrict viral access to the brain. In vitro models that recapitulate many of the properties of the human BBB endothelium are lacking, particularly with respect to the unique cellular and immunological mechanisms by which these cells restrict viral infections of the brain. Here, we developed a three-dimensional cell culture model that recapitulates many of the morphological and functional properties of the BBB microvasculature and apply this model to the study of RNA virus infections. The model we describe can therefore be used to study a variety of aspects of BBB physiology, including the mechanisms by which viruses might access the CNS, and could be used for the development and screening of antiviral therapeutics to limit this important step in viral pathogenesis. The blood-brain barrier (BBB) comprises the foremost protective barrier in the brain and is composed in part of a layer of microvascular endothelial cells that line the capillaries surrounding the brain. Here, we describe a human three-dimensional (3-D) cell-based model of the BBB microvascular endothelium that recapitulates properties of these cells in vivo, including physiologically relevant transcriptional profiles, the capacity to induce potent antimicrobial innate immune signaling, and the ability to resist infection by diverse RNA viruses, including members of the enterovirus (coxsackievirus B, echovirus 11, enterovirus 71, poliovirus) and flavivirus (dengue virus, Zika virus [ZIKV]) families. We show that disruption of apical tight junctions by proinflammatory cytokine tumor necrosis factor alpha (TNF-α) sensitizes 3-D-cultured BBB cells to ZIKV infection and that 3-D derived BBB cells can be used to model the transmigration of ZIKV-infected monocytes across the endothelial barrier to access underlying astrocytes. Taken together, our findings show that human BBB microvascular endothelial cells cultured in 3-D can be used to model the mechanisms by which RNA viruses access the central nervous system (CNS), which could be used for the development and screening of therapeutics to limit this event. IMPORTANCE Neurotropic viral infections are significant sources of global morbidity and mortality. The blood-brain barrier (BBB) is composed in part of a layer of microvascular endothelial cells and functions to restrict viral access to the brain. In vitro models that recapitulate many of the properties of the human BBB endothelium are lacking, particularly with respect to the unique cellular and immunological mechanisms by which these cells restrict viral infections of the brain. Here, we developed a three-dimensional cell culture model that recapitulates many of the morphological and functional properties of the BBB microvasculature and apply this model to the study of RNA virus infections. The model we describe can therefore be used to study a variety of aspects of BBB physiology, including the mechanisms by which viruses might access the CNS, and could be used for the development and screening of antiviral therapeutics to limit this important step in viral pathogenesis.