Polarized glycoprotein targeting affects the spread of measles virus in vitro and in vivo
Polarized glycoprotein targeting affects the spread of measles virus in vitro and in vivo
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DOI：
10.1099/vir.0.19663-0
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发表时间：
2004-04-01
影响因子：
3.8
通讯作者：
Maisner, A
中科院分区：
文献类型：
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作者：
Moll, M;Pfeuffer, J;Maisner, A
我们以前已经证明，质粒编码的麻疹病毒(MV)F和H蛋白的碱侧靶向依赖于糖蛋白胞浆尾部的单个酪氨酸残基，并且对于极化上皮细胞的融合活性是必不可少的。在这里，我们提供了关于极化糖蛋白表达对于感染性MV在培养中的细胞病变特性和体内致病机制的功能重要性的数据。通过引入单点突变，我们产生了重组病毒，其中一个或两个糖蛋白的碱侧靶向信号被破坏(酪氨酸突变体)。因此，突变的糖蛋白主要表达在极化的Madin-Darby犬肾细胞的顶膜上。与亲本MV不同的是，这些病毒突变体都不能在极化细胞中通过合胞体的形成传播，这表明两种MV糖蛋白都存在于细胞基侧表面，这是体外细胞间融合所必需的。使用棉鼠作为允许MV在呼吸道复制的动物模型，我们表明基侧糖蛋白靶向对于体内感染的传播也是重要的。而亲本MV能够在呼吸道上皮内横向传播，并从那里传播到下层组织中的细胞，而酪氨酸突变只感染单个上皮细胞和极少数上皮下细胞。这些数据有力地表明，MV糖蛋白的基侧靶向有助于克服上皮屏障，从而促进MV感染在体内的系统性传播。
We have shown previously that basolateral targeting of plasmid-encoded measles virus (MV) F and H protein is dependent on single tyrosine residues in the cytoplasmic tails of the glycoproteins and is essential for fusion activity in polarized epithelial cells. Here, we present data on the functional importance of polarized glycoprotein expression for the cytopathic properties of infectious MV in culture and for pathogenesis in vivo. By the introduction of single point mutations, we generated recombinant viruses in which the basolateral targeting signal of either one or both glycoproteins was destroyed (tyrosine mutants). As a consequence, the mutated glycoproteins were predominantly expressed on the apical membrane of polarized Madin-Darby canine kidney cells. In contrast to parental MV, none of these virus mutants was able to spread by syncytia formation in polarized cells showing that the presence of both MV glycoproteins at the basolateral cell surface is required for cell-to-cell fusion in vitro. Using cotton rats as an animal model that allows MV replication in the respiratory tract, we showed that basolateral glycoprotein targeting is also of importance for the spread of infection in vivo. Whereas parental MV was able to spread laterally within the respiratory epithelium and from there to cells in the underlying tissue, tyrosine mutants infected only single epithelial and very few subepithelial cells. These data strongly suggest that basolateral targeting of MV glycoproteins helps to overcome the epithelial barrier and thereby facilitates the systemic spread of MV infection in vivo.