Autophagy Benefits the Replication of Newcastle Disease Virus in Chicken Cells and Tissues
Autophagy Benefits the Replication of Newcastle Disease Virus in Chicken Cells and Tissues
复制标题
自噬有利于新城疫病毒在鸡细胞和组织中的复制
DOI：
10.1128/jvi.01849-13
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发表时间：
2014-01-01
影响因子：
5.4
通讯作者：
Ding, Chan
中科院分区：
文献类型：
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作者：
Sun, Yingjie;Yu, Shengqing;Ding, Chan
【摘要】：新城疫病毒(NDV)是一种重要的禽类病原体。我们之前报道过 NDV 会触发 U251 神经胶质瘤细胞的自噬，从而增强病毒复制。在这项研究中，我们研究了新城疫病毒是否会触发鸡细胞和组织中的自噬以增强病毒复制。我们证明，NDV 感染在鸡源性 DF-1 细胞和原代鸡胚成纤维细胞 (CEF) 细胞中诱导稳态自噬，这通过双膜或单膜囊泡的增加、绿色荧光蛋白 (GFP)-LC3 点的积累以及 LC3-I 向 LC3-II 的转化来证明。此外，我们通过监测p62/SQSTM1降解、LC3-II周转以及GFP-LC3溶酶体递送和蛋白水解来测量自噬通量，以证实NDV感染诱导了完整的自噬过程。药物抑制剂和 RNA 干扰对自噬的抑制减少了病毒复制，表明自噬在 NDV 感染中发挥着重要作用。此外，我们还进行了体内实验，观察了NDV感染鸡的心脏、肝脏、脾脏、肺和肾脏中LC3-I向LC3-II的转化。使用渥曼青霉素、氯喹或饥饿治疗诱导自噬的调节会影响肺和肠组织中新城疫病毒的产生和发病机制；然而，用雷帕霉素（一种哺乳动物细胞自噬诱导剂）处理，鸡细胞和组织中没有显示出可检测到的变化。此外，给予自噬抑制剂渥曼青霉素可提高NDV感染鸡的存活率。我们的研究提供了强有力的证据，表明新城疫病毒感染会诱导自噬，从而有利于新城疫病毒在鸡细胞和组织中的复制。
ABSTRACT Newcastle disease virus (NDV) is an important avian pathogen. We previously reported that NDV triggers autophagy in U251 glioma cells, resulting in enhanced virus replication. In this study, we investigated whether NDV triggers autophagy in chicken cells and tissues to enhance virus replication. We demonstrated that NDV infection induced steady-state autophagy in chicken-derived DF-1 cells and in primary chicken embryo fibroblast (CEF) cells, evident through increased double- or single-membrane vesicles, the accumulation of green fluorescent protein (GFP)-LC3 dots, and the conversion of LC3-I to LC3-II. In addition, we measured autophagic flux by monitoring p62/SQSTM1 degradation, LC3-II turnover, and GFP-LC3 lysosomal delivery and proteolysis, to confirm that NDV infection induced the complete autophagic process. Inhibition of autophagy by pharmacological inhibitors and RNA interference reduced virus replication, indicating an important role for autophagy in NDV infection. Furthermore, we conducted in vivo experiments and observed the conversion of LC3-I to LC3-II in heart, liver, spleen, lung, and kidney of NDV-infected chickens. Regulation of the induction of autophagy with wortmannin, chloroquine, or starvation treatment affects NDV production and pathogenesis in tissues of both lung and intestine; however, treatment with rapamycin, an autophagy inducer of mammalian cells, showed no detectable changes in chicken cells and tissues. Moreover, administration of the autophagy inhibitor wortmannin increased the survival rate of NDV-infected chickens. Our studies provide strong evidence that NDV infection induces autophagy which benefits NDV replication in chicken cells and tissues.