Cell-to-Cell Transmission of Turkey Herpesvirus in Chicken Embryo Cells via Tunneling Nanotubes
Cell-to-Cell Transmission of Turkey Herpesvirus in Chicken Embryo Cells via Tunneling Nanotubes
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DOI：
10.1637/aviandiseases-d-21-00022
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发表时间：
2021-09-01
期刊：
影响因子：
1.4
通讯作者：
Oishi, Eiji
中科院分区：
文献类型：
--
作者：
Okura, Takashi;Taneno, Akira;Oishi, Eiji
马立克氏病病毒（MDV）是一种致癌的甲疱疹病毒，在感染的鸡中引起免疫抑制、T细胞淋巴瘤和神经病变。为了保护鸡不受MDV感染，一种无毒的火鸡疱疹病毒活疫苗（HVT）已在世界范围内成功地用于鸡。许多疫苗制造商使用鸡胚成纤维细胞（CEF）生产HVT疫苗。一般认为，HVT是一种高度细胞相关的疱疹病毒，通过细胞间接触传播，但目前尚不清楚HVT如何从感染细胞传播到未感染的靶细胞。在这里，我们通过免疫荧光分析显示，从感染CEF细胞中观察到含有肌动蛋白细胞骨架和HVT抗原的纳米管与邻近细胞接触。当用肌动蛋白聚合或解聚抑制剂处理感染细胞时，来自感染细胞的纳米管的形成和延伸被极大地抑制，细胞间接触被消除，导致斑块形成和细胞相关病毒滴度急剧降低。我们的数据表明，通过由肌动蛋白丝组成的纳米管进行细胞间接触对于有效的病毒传播和复制至关重要。这一发现可能有助于进一步提高高效的HVT疫苗生产。
Marek's disease virus (MDV) is an oncogenic alphaherpesvirus that causes immunosuppression, T cell lymphomas, and neuropathic disease in infected chickens. To protect chickens from MDV infection, an avirulent live vaccine of turkey herpesvirus (HVT) has been successfully used in chickens worldwide. Many vaccine manufacturers have used chicken embryo fibroblast (CEF) cells to produce the HVT vaccine. Generally, it has been suggested that HVT is a highly cell-associated herpesvirus that spread via cell-to-cell contact, but it is unclear how HVT is transmitted from infected cells to uninfected target cells. Here, we show via immunofluorescence analysis that nanotubes containing the actin cytoskeleton and HVT antigens from infected CEF cells were observed to contact neighboring cells. When the infected cells were treated with inhibitors for actin polymerization or depolymerization, the formation and extension of the nanotubes from infected cells were greatly inhibited and the intercellular contact was abolished, leading to a drastic reduction in plaque formation and viral titers of the cell-associated virus. Our data indicate that cell-to-cell contacts via nanotubes composed of actin filaments are essential for efficient viral spreading and replication. This finding might contribute to the further improvement of efficient HVT vaccine production.