Respiratory Syncytial Virus Uses CX3CR1 as a Receptor on Primary Human Airway Epithelial Cultures.
Respiratory Syncytial Virus Uses CX3CR1 as a Receptor on Primary Human Airway Epithelial Cultures.
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DOI：
10.1371/journal.ppat.1005318
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发表时间：
2015-12
期刊：
影响因子：
6.7
通讯作者：
Peeples ME
中科院分区：
文献类型：
--
作者：
Johnson SM;McNally BA;Ioannidis I;Flano E;Teng MN;Oomens AG;Walsh EE;Peeples ME
呼吸道合胞病毒（RSV）是婴儿下呼吸道疾病的最常见原因，但没有疫苗或有效的治疗方法。永生化细胞的RSV感染的起始在很大程度上依赖于细胞表面硫酸乙酰肝素（HS），其是永生化细胞中RSV附着（G）糖蛋白的受体。然而，RSV通过顶端表面感染原代分化良好的人气道上皮（HAE）培养物中的纤毛细胞，但在该表面上检测不到HS。在这里，我们表明，可溶性HS抑制永生化细胞的感染，但不是HAE文化，确认HS不是HAE文化的受体。相反，不阻断RSV感染永生化细胞的针对G蛋白的“非中和”单克隆抗体确实抑制HAE培养物的感染。该抗体先前显示阻断G蛋白和趋化因子受体CX3CR1之间的相互作用，并且我们已经将该抗体的结合位点映射到G蛋白中的CX3C基序及其周围区域。我们发现，CX3CR1是目前在HAE文化的纤毛细胞的顶端表面，特别是在纤毛。HAE培养物的RSV感染通过针对CX3CR1的抗体和G蛋白CX3C基序中的突变而减少。此外，缺乏CX3CR1的小鼠对RSV感染的敏感性较低。这些发现表明，RSV使用CX3CR1作为HAE培养物上的细胞受体，并强调了使用生理学相关模型研究病毒进入和抗体中和的重要性。呼吸道合胞病毒（RSV）是世界范围内婴儿死亡的第二大常见传染性原因。尽管有这种巨大的临床影响，但没有有效的抗病毒药物或针对RSV的疫苗。在此，我们发现RSV附着（G）糖蛋白使用CX3CR1作为原代人气道上皮（HAE）培养物（RSV感染人肺的极好模型）上的受体。G蛋白含有CX3C基序，我们发现该区域对于其在HAE培养物感染中的作用至关重要，但对于永生化细胞则不然。此外，我们发现针对G蛋白的抗体中和HAE培养物的RSV感染不同于永生化细胞。这些见解表明，HAE培养物应用于定量中和抗体，包括在疫苗开发期间，CX3CR1与RSV G蛋白的相互作用可能是抗病毒药物开发的靶点，并且应考虑将G蛋白纳入疫苗中。
Respiratory syncytial virus (RSV) is the most frequent cause of lower respiratory disease in infants, but no vaccine or effective therapy is available. The initiation of RSV infection of immortalized cells is largely dependent on cell surface heparan sulfate (HS), a receptor for the RSV attachment (G) glycoprotein in immortalized cells. However, RSV infects the ciliated cells in primary well differentiated human airway epithelial (HAE) cultures via the apical surface, but HS is not detectable on this surface. Here we show that soluble HS inhibits infection of immortalized cells, but not HAE cultures, confirming that HS is not the receptor on HAE cultures. Conversely, a “non-neutralizing” monoclonal antibody against the G protein that does not block RSV infection of immortalized cells, does inhibit infection of HAE cultures. This antibody was previously shown to block the interaction between the G protein and the chemokine receptor CX3CR1 and we have mapped the binding site for this antibody to the CX3C motif and its surrounding region in the G protein. We show that CX3CR1 is present on the apical surface of ciliated cells in HAE cultures and especially on the cilia. RSV infection of HAE cultures is reduced by an antibody against CX3CR1 and by mutations in the G protein CX3C motif. Additionally, mice lacking CX3CR1 are less susceptible to RSV infection. These findings demonstrate that RSV uses CX3CR1 as a cellular receptor on HAE cultures and highlight the importance of using a physiologically relevant model to study virus entry and antibody neutralization. Respiratory syncytial virus (RSV) is the second most common infectious cause of infant death worldwide. Despite this great clinical impact, no effective antivirals or vaccines against RSV are available. Here we find that the RSV attachment (G) glycoprotein uses CX3CR1 as a receptor on primary human airway epithelial (HAE) cultures, an excellent model of RSV infection of the human lung. The G protein contains a CX3C motif and we find that this region is critical for its role in infection of HAE cultures, but not of immortalized cells. Furthermore, we find that antibodies against the G protein neutralize RSV infection of HAE cultures differently from immortalized cells. These insights suggest that HAE cultures should be used to quantify neutralizing antibodies, including during vaccine development, that the CX3CR1 interaction with the RSV G protein could be a target for antiviral drug development, and that the G protein should be considered for inclusion in vaccines.