Transcellular transport of West Nile virus-like particles across human endothelial cells depends on residues 156 and 159 of envelope protein

BMC Microbiology

Research Article

Manabu Igarashi¹  Tadaki Suzuki²  Satoko Yamanouchi²  Takashi Kimura²  Hirofumi Sawa³  Yoshinori Makino³  Rie Hasebe⁴  Akihiko Maeda⁵  Motohiro Horiuchi⁵ 

[1] Department of Global Epidemiology, Hokkaido University Research Center for Zoonosis Control, Kita 20, Nishi 10, Kita-ku, 001-0020, Sapporo, Japan;Department of Molecular Pathobiology, Hokkaido University Research Center for Zoonosis Control, Kita 20, Nishi 10, Kita-ku, 001-0020, Sapporo, Japan;Department of Molecular Pathobiology, Hokkaido University Research Center for Zoonosis Control, Kita 20, Nishi 10, Kita-ku, 001-0020, Sapporo, Japan;Global COE program, Hokkaido University, Japan;Department of Molecular Pathobiology, Hokkaido University Research Center for Zoonosis Control, Kita 20, Nishi 10, Kita-ku, 001-0020, Sapporo, Japan;Laboratory of Veterinary Hygiene, Graduate School of Veterinary Medicine, Hokkaido University, Kita 18, Nishi 9, Kita-ku, 060-0818, Sapporo, Japan;Laboratory of Veterinary Hygiene, Graduate School of Veterinary Medicine, Hokkaido University, Kita 18, Nishi 9, Kita-ku, 060-0818, Sapporo, Japan;

关键词: Tight Junction;    Human Umbilical Vein Endothelial Cell;    West Nile Virus;    Human Endothelial Cell;    West Nile Virus Infection;   

DOI  :  10.1186/1471-2180-10-165

 received in 2010-01-13, accepted in 2010-06-08,  发布年份 2010

来源: Springer

【 摘 要 】

BackgroundWest 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.Results6-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.ConclusionOur 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.

【 授权许可】

CC BY   
© Hasebe et al; licensee BioMed Central Ltd. 2010