Identification of a putative coreceptor on vero cells that participates in dengue 4 virus infection
Identification of a putative coreceptor on vero cells that participates in dengue 4 virus infection
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DOI：
10.1128/jvi.75.17.7818-7827.2001
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发表时间：
2001-09-01
影响因子：
5.4
通讯作者：
Del Angel, RM
中科院分区：
文献类型：
--
作者：
Martínez-Barragán, JD;Del Angel, RM
登革病毒通过位于病毒膜表面的包膜（E）糖蛋白附着于细胞表面受体来感染靶细胞。在Vero和BHK细胞上，蛋白聚糖的硫酸乙酰肝素（HS）部分是登革病毒的受体;然而，其他蛋白质也被描述为C6/36、HL 60和BM细胞上的推定登革病毒受体。HS还可以作为其他类型病毒的受体或作为需要额外宿主细胞分子以允许病毒渗透的病毒的附着分子。在这项研究中，我们寻找分子以外的HS，可以参与登革病毒感染的Vero细胞。标记的登革4型病毒与74和44 kDa的两种分子以高亲和力结合。登革病毒与74 kDa分子的结合对蛋白酶和高碘酸钠处理敏感，对肝素酶处理具有抗性。凝集素，如伴刀豆球蛋白A和麦胚凝集素防止登革热病毒结合的74-和44-kDa的蛋白质在覆盖测定，而植物血凝素P不影响结合，这表明碳水化合物残基（α-甘露糖或N-乙酰葡糖胺）是重要的病毒结合宿主细胞。蛋白酶敏感性，生物素标记，免疫荧光与多克隆抗体提出了对74 kDa的蛋白质一致确定的蛋白质的Vero细胞表面上。此外，针对74-kDa蛋白的抗体能够抑制登革病毒感染。这些数据表明，HS可能作为一个主要的受体，可能集中在Vero细胞表面的病毒颗粒，然后其他分子，如74 kDa的蛋白质，可能参与作为辅助受体在病毒的渗透。74-kDa蛋白可能构成登革热病毒感染Vero细胞的假定受体复合物的一部分。
Dengue virus infects target cells by attaching to a cell surface receptor through the envelope (E) glycoprotein, located on the surface of the viral membrane. On Vero and BHK cells, heparan sulfate (HS) moieties of proteoglycans are the receptors for dengue virus; however, additional proteins have also been described as putative dengue virus receptors on C6/36, HL60, and BM cells. HS can also act as a receptor for other types of viruses or as an attachment molecule for viruses that require additional host cell molecules to allow viral penetration. In this study we searched for molecules other than HS that could participate in dengue virus infection of Vero cells. Labeled dengue 4 virus bound with high affinity to two molecules of 74 and 44 kDa. Binding of dengue virus to the 74-kDa molecule was susceptible to protease and sodium periodate treatment and resistant to heparinase treatments. Lectins such as concanavalin A and wheat germ agglutinin prevented dengue virus binding to both the 74- and the 44-kDa protein in overlay assays, while phytohemagglutinin P did not affect binding, suggesting that carbohydrate residues (a-mannose or N-acetylglucosamine) are important in virus binding to host cells. Protease susceptibility, biotin labeling, and immunofluorescence with a polyclonal antibody raised against the 74-kDa protein consistently identified the protein on the surfaces of Vero cells. Moreover, the antibody against the 74-kDa protein was able to inhibit dengue virus infection. These data suggest that HS might serve as a primary receptor, probably concentrating virus particles on the surfaces of Vero cells, and then other molecules, such as the 74-kDa protein, might participate as coreceptors in viral penetration. The 74-kDa protein possibly constitutes part of a putative receptor complex for dengue virus infection of Vero cells.