Lack of Norovirus Replication and Histo-Blood Group Antigen Expression in 3-Dimensional Intestinal Epithelial Cells
Published Date:Mar 2013
Source:Emerg Infect Dis. 19(3):431-438.
Keywords:3-D Intestinal Epithelial Cell Culture
Blood Group Antigens
Blood Group H Type Antigens
Cell Culture Techniques
Human Intestinal Epithelial Cells
Lewis Blood-group System
Real-Time Polymerase Chain Reaction
Rotating Wall Vessel Bioreactor
Viral Nonstructural Proteins
Viral Structural Proteins
Description: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.
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