Transcellular transport of West Nile virus-like particles across human endothelial cells depends on residues 156 and 159 of envelope protein.
Transcellular transport of West Nile virus-like particles across human endothelial cells depends on residues 156 and 159 of envelope protein.
复制标题
DOI：
10.1186/1471-2180-10-165
复制
发表时间：
2010-06-08
期刊：
影响因子：
4.2
通讯作者：
Kimura T
中科院分区：
文献类型：
--
作者：
Hasebe R;Suzuki T;Makino Y;Igarashi M;Yamanouchi S;Maeda A;Horiuchi M;Sawa H;Kimura T
西尼罗病毒（WNV）经蚊虫叮咬侵入宿主后引起病毒血症。内皮细胞可能在西尼罗河病毒从血流扩散到中枢神经系统和外周组织中发挥重要作用。在这里，我们分析了高毒力的NY99 6-LP菌株（6-LP VLPs）和低毒力的Eg101菌株（Eg VLPs）的病毒样颗粒（VLPs）对交叉培养的人内皮细胞的能力。6-LP VLPs从内皮细胞的根尖向基底外侧运输，而Eg VLPs几乎没有运输。在6-LP VLPs的运输过程中，紧密连接标记ZO-1的定位和紧密连接的完整性未受到损害。6-LP VLPs的运输被filipin抑制，filipin可以阻止胆固醇依赖性膜筏的形成，提示筏相关的膜运输参与其中。为了确定负责VLPs运输的氨基酸残基，我们制作了VLPs突变体，其中E蛋白残基在6-LP和Eg菌株之间交换。残基156和159的双氨基酸取代极大地破坏了VLPs的运输。我们的研究结果表明，跨细胞途径与6-LP VLPs转运有关。我们还发现残基156和159的组合在VLPs跨内皮细胞的运输中起重要作用。
West Nile virus (WNV) causes viremia after invasion to the hosts by mosquito bite. Endothelial cells could play an important role in WNV spread from the blood stream into the central nervous system and peripheral tissues. Here, we analyzed the capacity of virus-like particles (VLPs) of the highly virulent NY99 6-LP strain (6-LP VLPs) and the low virulence Eg101 strain (Eg VLPs) to cross cultured human endothelial cells. 6-LP VLPs were transported from the apical to basolateral side of endothelial cells, whereas Eg VLPs were hardly transported. The localization of tight junction marker ZO-1 and the integrity of tight junctions were not impaired during the transport of 6-LP VLPs. The transport of 6-LP VLPs was inhibited by treatment with filipin, which prevents the formation of cholesterol-dependent membrane rafts, suggesting the involvement of raft-associated membrane transport. To determine the amino acid residues responsible for the transport of VLPs, we produced mutant VLPs, in which residues of E protein were exchanged between the 6-LP and Eg strains. Double amino acid substitution of the residues 156 and 159 greatly impaired the transport of VLPs. Our results suggest that a transcellular pathway is associated with 6-LP VLPs transport. We also showed that the combination of the residues 156 and 159 plays an important role in the transport of VLPs across endothelial cells.
登录
查看更多内容
影响因子：
11.8
作者：
Asnis, DS;Conetta, R;Sampson, BA
通讯作者：
Sampson, BA
影响因子：
2.7
作者：
HALEVY, M;AKOV, Y;LUSTIG, S
通讯作者：
LUSTIG, S
影响因子：
3.7
作者：
Beasley, DWC;Li, L;Barrett, ADT
通讯作者：
Barrett, ADT
影响因子：
6.4
作者：
Armah, Henry B.;Wang, Guoji;Wiley, Clayton A.
通讯作者：
Wiley, Clayton A.
影响因子：
3.7
作者：
Chu, JJH;Leong, PWH;Ng, ML
通讯作者：
Ng, ML