Japanese encephalitis virus infects porcine kidney epithelial PK15 cells via clathrin- and cholesterol-dependent endocytosis.
BACKGROUND
Japanese encephalitis virus (JEV) is a mosquito-borne flavivirus that causes acute viral encephalitis in humans. Pigs are important amplifiers of JEV. The entry mechanism of JEV into porcine cells remains largely unknown. In this study, we present a study of the internalization mechanism of JEV in porcine kidney epithelial PK15 cells.


RESULTS
We demonstrated that the disruption of the lipid raft by cholesterol depletion with methyl-β-cyclodextrin (MβCD) reduced JEV infection. We also found that the knockdown of clathrin by small interfering RNA (siRNA) significantly reduced JEV-infected cells and JEV E-glycoprotein levels, suggesting that JEV utilizes clathrin-dependent endocytosis. In contrast, the knockdown of caveolin-1, a principal component of caveolae, had only a small (although statistically significant) effect on JEV infection, however, JEV entry was not affected by genistein. These results suggested that JEV entry was independent of caveolae.


CONCLUSIONS
Taken together, our results demonstrate that JEV enters porcine kidney epithelial PK15 cells through cholesterol- and clathrin-mediated endocytosis.
Key Laboratory of Agricultural Animal Genetics, Breeding and Reproduction, Ministry of Education & College of Animal Science and Technology, Huazhong Agricultural University, Wuhan 430070, China.
Songbai
S
Yang
Minhui
M
He
Xiangdong
X
Liu
Xinyun
X
Li
Bin
B
Fan
Shuhong
S
Zhao
D000818:Animals; D002460:Cell Line; D002784:Cholesterol / Q000378:metabolism; D002966:Clathrin / Q000378:metabolism*; D004664:Encephalitis Virus, Japanese / Q000502:physiology*; D004705:Endocytosis; D020868:Gene Silencing; D013552:Swine; D053586:Virus Internalization
D002966:Clathrin; D002784:Cholesterol