Bile acids accumulate norovirus-like particles and enhance binding to and entry into human enteric epithelial cells.

Human norovirus (HuNoV) is a leading cause of acute viral gastroenteritis, but despite high impact on public health and healthcare, the mechanisms of viral attachment to and entry into target cells are not yet fully understood. Recent reports indicate that saliva and bile contribute to the transmission of HuNoV. For example, human bile acids increase cell surface ceramide levels in human enteroids, which improves norovirus entry into cells, resulting in enhanced replication. Bile acids can also interact directly with the norovirus capsid, but it is not known whether bile or other gastrointestinal body fluids directly affect HuNoV attachment to host cells. In this study, we investigated the effects of patient-derived gastric juice, pancreatic juice, and bile on HuNoV GII.4 virus-like particle (VLP) attachment to and entry into a human duodenal cell line, HuTu-80. We show that while gastric juice and pancreatic juice do not affect viral attachment or entry, bile-in particular, hydrophobic bile acids-significantly enhance cellular attachment and subsequent entry of GII.4 VLPs into cells. In addition, we show that hydrophobic bile acids induce the accumulation of viral particles in the vicinity of cells. These results suggest the presence of a new