Analysis and optimization of a Caco-2 cell culture model for infection with human norovirus

Human noroviruses (hNoVs) are an important cause of acute gastroenteritis worldwide. However, the lack of a reproducible in vitro cell culture system has impaired research and the development of preventive measures, therapeutic drugs, and vaccines. The aim of this study was to analyze and optimize a suitable cell line for in vitro cultivation of hNoV. The Caco-2 cell line, which is of colorectal origin and differentiates spontaneously into intestinal enterocyte-like cells, was chosen as a model. It was found that differentiated cells were more susceptible to infection with hNoV, resulting in a higher virus yield. This was accompanied by an increase in H type 1 antigen in the cell membrane during differentiation, which functions as an attachment factor for hNoV. Induced overexpression of H type 1 antigen in undifferentiated Caco-2 cells resulted in an increase in viral output to a level similar to that in differentiated cells. However, the relatively low level of viral output, which contrasts with what is observed in vivo, shows that the viral replication cycle is restricted in this model. The results indicate that there is a block at the level of viral release.