Characterization of plasma membrane-associated proteins from Aedes albopictus mosquito (C6/36) cells that mediate West Nile virus binding and infection
Characterization of plasma membrane-associated proteins from Aedes albopictus mosquito (C6/36) cells that mediate West Nile virus binding and infection
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DOI：
10.1016/j.virol.2005.05.026
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发表时间：
2005-09-01
期刊：
影响因子：
3.7
通讯作者：
Ng, ML
中科院分区：
文献类型：
--
作者：
Chu, JJH;Leong, PWH;Ng, ML
本研究从白纹伊蚊（C6/36）细胞中分离并鉴定了西尼罗病毒（WNV）的假定受体分子。WNV与C6/36细胞的结合以5000个颗粒/细胞饱和。当用蛋白酶K预处理时，WNV进入C6/36细胞被强烈抑制，在较小程度上用高碘酸钠预处理。然而，用磷脂酶、糖苷酶、肝素酶和神经酰亚胺酶预处理C6/36细胞对病毒进入没有影响。通过病毒覆盖蛋白印迹分析，观察到WNV与分离自C6/36细胞的140-kDa、95-kDa、70-kDa和55-kDa质膜相关分子结合。针对95-kDa和70-kDa膜蛋白产生的鼠抗体有效地阻断了C6/36细胞中的WNV、日本脑炎病毒（JEV）和登革病毒（DV）血清型2感染。此外，重组WNV包膜结构域III蛋白与C6/ 36细胞的结合可被抗95-kDa和抗70-kDa膜蛋白抗体抑制。这些数据有力地支持了95-kDa和70-kDa质膜相关蛋白是蚊媒黄病毒（WNV、JEV和DV）在蚊子细胞上的受体复合物的一部分的可能性。(c)2005年爱思唯尔公司All rights reserved.
This study isolated and characterized the West Nile virus (WNV) putative receptor molecule(s) from Aedes albopictus mosquito (C6/36) cells. The binding of WNV to C6/36 cells was saturated with 5000 particles per cell. The entry of WNV into C6/36 cells was strongly inhibited when pretreated with protemase K and to a lesser extent with sodium periodate. However, pretreatment of C6/36 cells with phospholipases, glycosidases, heparinases and neurimidase had no effect on virus entry. By using virus overlay protein blot assay, WNV was observed to bind to the 140-kDa, 95-kDa, 70-kDa and 55-kDa plasma membrane-associated molecules isolated from C6/36 cells. Murine antibodies generated against the 95-kDa and 70-kDa membrane proteins effectively blocked WNV, Japanese encephalitis virus (JEV) and Dengue virus (DV) serotype 2 infection in C6/36 cells. In addition, the binding of the recombinant-WNV envelope domain III protein to C6/ 36 cells can be inhibited by the anti-95-kDa and anti-70-kDa membrane protein antibodies. These data strongly supported the possibility that the 95-kDa and 70-kDa plasma membrane-associated proteins are part of a receptor complex for mosquito-borne flaviviruses (WNV, JEV and DV) on mosquito cells. (c) 2005 Elsevier Inc. All rights reserved.