Newcastle Disease Virus Entry into Chicken Macrophages via a pH-Dependent, Dynamin and Caveola-Mediated Endocytic Pathway That Requires Rab5
Newcastle Disease Virus Entry into Chicken Macrophages via a pH-Dependent, Dynamin and Caveola-Mediated Endocytic Pathway That Requires Rab5
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DOI：
10.1128/jvi.02288-20
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发表时间：
2021-03
影响因子：
5.4
通讯作者：
Ran Zhao;Q. Shi;Zong-xi Han;Z. Fan;Hui Ai;Linna Chen;Le Li;Tianyi Liu;Junfeng Sun;Shengwang Liu
中科院分区：
文献类型：
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作者：
Ran Zhao;Q. Shi;Zong-xi Han;Z. Fan;Hui Ai;Linna Chen;Le Li;Tianyi Liu;Junfeng Sun;Shengwang Liu
虽然纽卡斯尔病毒（NDV）的致病机理已被广泛研究，但NDV进入宿主细胞的详细机制仍不清楚。巨噬细胞是宿主抵御病原体感染的第一线防御者。摘要纽卡斯尔病毒（NDV）进入细胞的途径和机制目前尚不清楚。在本研究中，我们证明，鸡干扰素诱导的跨膜蛋白1（chIFITM 1），这是位于早期的内涵体，可以限制NDV在鸡巨噬细胞系HD 11中的复制，这表明NDV的内吞进入鸡巨噬细胞。在此基础上，我们对NDV进入鸡巨噬细胞的机制进行了系统的研究。首先，我们证明了在NDV进入过程中需要低pH条件和发动蛋白。然而，NDV进入鸡巨噬细胞是独立的网格蛋白介导的内吞作用。我们还发现NDV的进入依赖于膜胆固醇。制霉菌素和佛波醇12-肉豆蔻酸酯13-乙酸酯处理，显性阴性（DN）小窝蛋白-1的过表达，或敲低小窝蛋白-1，显着减少NDV的进入和复制，表明NDV的进入依赖于小窝介导的内吞作用。然而，巨胞饮并没有发挥作用，在新城疫病毒进入鸡巨噬细胞。此外，我们发现Rab 5而不是Rab 7参与了NDV的进入和传播。NDV与Rab 5和早期内体共定位表明NDV病毒粒子在内化后以Rab 5依赖的方式转运到早期内体。特别值得注意的是，小窝介导的内吞作用也被NDV用于进入原代鸡巨噬细胞。此外，NDV通过不同的途径进入不同的细胞类型。总的来说，我们的研究结果首次表明，NDV病毒粒子进入鸡巨噬细胞通过pH值依赖性，发动蛋白和小窝介导的内吞途径和Rab 5参与NDV的交通和位置。虽然纽卡斯尔病病毒（NDV）的发病机制已被广泛研究，但NDV进入宿主细胞的详细机制在很大程度上是未知的。巨噬细胞是宿主抵御病原体感染的第一线防御者。鸡巨噬细胞被认为是NDV感染过程中的主要靶细胞类型之一。本文对NDV进入鸡巨噬细胞的机制进行了全面的研究。这是第一份证明NDV通过pH依赖性、发动蛋白和小窝介导的需要Rab 5的内吞途径进入鸡巨噬细胞的报告。这一结果对于我们了解NDV进入鸡巨噬细胞的机制具有重要意义，将进一步加深对NDV致病机制的认识，并为开发新的NDV感染的预防或治疗策略提供有用的线索。此外，这些信息将有助于我们进一步了解副粘病毒科Avulavirus属其他成员的发病机制。
Although the pathogenesis of Newcastle disease virus (NDV) has been extensively studied, the detailed mechanism of NDV entry into host cells is largely unknown. Macrophages are the first-line defenders of host defense against infection of pathogens. ABSTRACT The cellular entry pathways and the mechanisms of Newcastle disease virus (NDV) entry into cells are poorly characterized. In this study, we demonstrated that chicken interferon-induced transmembrane protein 1 (chIFITM1), which is located in the early endosomes, could limit the replication of NDV in chicken macrophage cell line HD11, suggesting the endocytic entry of NDV into chicken macrophages. Then, we presented a systematic study about the entry mechanism of NDV into chicken macrophages. First, we demonstrated that a low-pH condition and dynamin were required during NDV entry. However, NDV entry into chicken macrophages was independent of clathrin-mediated endocytosis. We also found that NDV entry was dependent on membrane cholesterol. The NDV entry and replication were significantly reduced by nystatin and phorbol 12-myristate 13-acetate treatment, overexpression of dominant-negative (DN) caveolin-1, or knockdown of caveolin-1, suggesting that NDV entry depends on caveola-mediated endocytosis. However, macropinocytosis did not play a role in NDV entry into chicken macrophages. In addition, we found that Rab5, rather than Rab7, was involved in the entry and traffic of NDV. The colocalization of NDV with Rab5 and early endosome suggested that NDV virion was transported to early endosomes in a Rab5-dependent manner after internalization. Of particular note, the caveola-mediated endocytosis was also utilized by NDV to enter primary chicken macrophages. Moreover, NDV entered different cell types using different pathways. Collectively, our findings demonstrate for the first time that NDV virion enters chicken macrophages via a pH-dependent, dynamin and caveola-mediated endocytosis pathway and that Rab5 is involved in the traffic and location of NDV. IMPORTANCE Although the pathogenesis of Newcastle disease virus (NDV) has been extensively studied, the detailed mechanism of NDV entry into host cells is largely unknown. Macrophages are the first-line defenders of host defense against infection of pathogens. Chicken macrophages are considered one of the main types of target cells during NDV infection. Here, we comprehensively investigated the entry mechanism of NDV in chicken macrophages. This is the first report to demonstrate that NDV enters chicken macrophages via a pH-dependent, dynamin and caveola-mediated endocytosis pathway that requires Rab5. The result is important for our understanding of the entry of NDV in chicken macrophages, which will further advance the knowledge of NDV pathogenesis and provide useful clues for the development of novel preventive or therapeutic strategies against NDV infection. In addition, this information will contribute to our further understanding of pathogenesis with regard to other members of the Avulavirus genus in the Paramyxoviridae family.