title: Asialoglycoprotein receptor facilitates infection of PLC/PRF/5 cells by HEV through interaction with ORF2.
abstract: Although the biological and epidemiological features of hepatitis E virus (HEV) have been studied extensively in recent years, the mechanism by which HEV infects cells is still poorly understood. In this study, coimmunoprecipitation, pull-down, and ELISA were used to show that the HEV ORF2 protein interacts directly with the ectodomain of both ASGR1 and ASGR2. Susceptibility to HEV correlated positively with the expression level of surface asialoglycoprotein receptor (ASGPR) in cell lines. ASGPR-directed small interfering RNA (siRNA) in HEV-infected PLC/PRF/5 cells had no significant effect on HEV release, suggesting that ASGPR mainly regulates the viral binding and entry steps. Both the purified ASGPR ectodomain and anti-ASGPR antibodies disturbed the binding of HEV to PLC/PRF/5 cells. The classic ASGPR ligands asialofetuin, asialoganglioside, and fibronectin competitively inhibited the binding of HEV to hepatocytes in the presence of calcium. HeLa cell lines stably expressing ASGPR displayed increased HEV-binding capacity, whereas ASGPR-knockout PLC/PRF/5 cell lines had lower HEV-binding capacity. Thus, our study demonstrates that ASGPR is involved in and facilitates HEV infection by binding to ORF2. J. Med. Virol. 88:2186-2195, 2016. © 2016 Wiley Periodicals, Inc.
authors.affiliation: Division of HIV/AIDS and Sexually-Transmitted Virus Vaccines, National Institutes for Food and Drug Control, Beijing, China.
authors.forename: Li
authors.initials: L
authors.lastname: Zhang
authors.affiliation: Division of HIV/AIDS and Sexually-Transmitted Virus Vaccines, National Institutes for Food and Drug Control, Beijing, China.
authors.forename: Yabin
authors.initials: Y
authors.lastname: Tian
authors.affiliation: Division of HIV/AIDS and Sexually-Transmitted Virus Vaccines, National Institutes for Food and Drug Control, Beijing, China.
authors.forename: Zhiheng
authors.initials: Z
authors.lastname: Wen
authors.affiliation: Division of Monoclonal Antibody Products, National Institutes for Food and Drug Control, Beijing, China.
authors.forename: Feng
authors.initials: F
authors.lastname: Zhang
authors.affiliation: Division of HIV/AIDS and Sexually-Transmitted Virus Vaccines, National Institutes for Food and Drug Control, Beijing, China.
authors.forename: Ying
authors.initials: Y
authors.lastname: Qi
authors.affiliation: Division of HIV/AIDS and Sexually-Transmitted Virus Vaccines, National Institutes for Food and Drug Control, Beijing, China.
authors.forename: Weijin
authors.initials: W
authors.lastname: Huang
authors.affiliation: Department of Bio-Diagnosis, Beijing Institute of Basic Medical Sciences, Beijing, China.
authors.forename: Heqiu
authors.initials: H
authors.lastname: Zhang
authors.affiliation: Division of HIV/AIDS and Sexually-Transmitted Virus Vaccines, National Institutes for Food and Drug Control, Beijing, China. wangyc@nifdc.org.cn, wangyc3@126.com.
authors.forename: Youchun
authors.initials: Y
authors.lastname: Wang
mesh_terms: D037263:Asialoglycoprotein Receptor / Q000378:metabolism*; D002460:Cell Line; D006367:HeLa Cells; D016752:Hepatitis E virus / Q000737:chemistry / Q000502:physiology*; D022781:Hepatocytes / Q000821:virology; D054884:Host-Pathogen Interactions; D006801:Humans; D014764:Viral Proteins / Q000378:metabolism*; D053585:Virus Attachment; D053586:Virus Internalization
chemical_list: C546927:ASGR1 protein, human; C546932:ASGR2 protein, human; D037263:Asialoglycoprotein Receptor; C400971:ORF2 protein, Hepatitis E virus; D014764:Viral Proteins
keywords: Hepatitis E virus; ORF2; asialoglycoprotein receptor; virus-host interaction