title: Lack of norovirus replication and histo-blood group antigen expression in 3-dimensional intestinal epithelial cells.
abstract: Noroviruses (NoVs) are a leading cause of gastroenteritis worldwide. An in vitro model for NoV replication remains elusive, making study of the virus difficult. A previous study, which used a 3-dimensional (3-D) intestinal model derived from INT-407 cells reported NoV replication and extensive cytopathic effects (CPE). Using the same 3-D model, but with highly purified Norwalk virus (NV), we attempted to replicate this study. Our results showed no evidence of NV replication by real-time PCR of viral RNA or by immunocytochemical detection of viral structural and nonstructural proteins. Immunocytochemical analysis of the 3-D cultures also showed no detectable presence of histo-blood group antigens that participate in NV binding and host tropism. To determine the potential cause of CPE observed in the previous study, we exposed 3-D cultures to lipopolysaccharide concentrations consistent with contaminated stool samples and observed morphologic features similar to CPE. We conclude that the 3-D INT-407 model does not support NV replication.
authors.affiliation: Department of Basic Medical Sciences, University of Arizona, Phoenix, AZ 85004-3902, USA. mherbst1@email.arizona.edu
authors.forename: Melissa M
authors.initials: MM
authors.lastname: Herbst-Kralovetz
authors.forename: Andrea L
authors.initials: AL
authors.lastname: Radtke
authors.forename: Margarita K
authors.initials: MK
authors.lastname: Lay
authors.forename: Brooke E
authors.initials: BE
authors.lastname: Hjelm
authors.forename: Alice N
authors.initials: AN
authors.lastname: Bolick
authors.forename: Shameema S
authors.initials: SS
authors.lastname: Sarker
authors.forename: Robert L
authors.initials: RL
authors.lastname: Atmar
authors.forename: David H
authors.initials: DH
authors.lastname: Kingsley
authors.forename: Charles J
authors.initials: CJ
authors.lastname: Arntzen
authors.forename: Mary K
authors.initials: MK
authors.lastname: Estes
authors.forename: Cheryl A
authors.initials: CA
authors.lastname: Nickerson
mesh_terms: D001789:Blood Group Antigens / Q000378:metabolism; D002449:Cell Aggregation; D018929:Cell Culture Techniques; D002460:Cell Line; D004847:Epithelial Cells / Q000276:immunology / Q000821:virology*; D005759:Gastroenteritis / Q000276:immunology / Q000473:pathology / Q000821:virology*; D006801:Humans; D007413:Intestinal Mucosa / Q000276:immunology / Q000473:pathology / Q000821:virology*; D008070:Lipopolysaccharides / Q000494:pharmacology; D029322:Norovirus / Q000502:physiology*; D012367:RNA, Viral / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D017361:Viral Nonstructural Proteins / Q000378:metabolism; D015678:Viral Structural Proteins / Q000378:metabolism; D056189:Viral Tropism; D014779:Virus Replication
chemical_list: D001789:Blood Group Antigens; D008070:Lipopolysaccharides; D012367:RNA, Viral; D017361:Viral Nonstructural Proteins; D015678:Viral Structural Proteins
keywords: 3-D intestinal epithelial cell culture; INT-407; Lewis blood-group system; Norwalk virus; blood group H type antigens; human intestinal epithelial cells; norovirus replication; rotating wall vessel bioreactor; viruses