biostudies-literatureUnknown99(11)Costantini VNorovirus is the leading cause of acute gastroenteritis worldwide. Since the development of human intestinal enteroid (HIE) cell culture systems, significant advancements have been made in exploring and evaluating therapeutic strategies to combat human norovirus infections. Here, we investigate whether 25-hydroxycholesterol (25-HC), a key regulator of cholesterol homeostasis and bile acid production, can restrict human norovirus replication in jejunal HIE monolayers. Jejunal HIE monolayers derived from two donors (J2 and J3 cell lines) were treated with increasing concentrations of 25-HC (ranging from 0.0001 µM to 5 µM) for 24 hours. Monolayers were then infected with norovirus GII.1[P33] or GII.4 Sydney[P31] and incubated for 24 hours at 37°C and 5% CO₂. Viral RNA was quantifieJournal of virologye0110925https://www.ebi.ac.uk/biostudies/studies/S-EPMC12645967biostudies-literature25-Hydroxycholesterol restricts human norovirus replication in human intestinal enteroids.PMC12645967Costantini VVinje JHuynh KMboko WPMontmayeur AMPark GWXu RChhabra PWall Hfalse25-Hydroxycholesterol restricts human norovirus replication in human intestinal enteroids.Norovirus is the leading cause of acute gastroenteritis worldwide. Since the development of human intestinal enteroid (HIE) cell culture systems, significant advancements have been made in exploring and evaluating therapeutic strategies to combat human norovirus infections. Here, we investigate whether 25-hydroxycholesterol (25-HC), a key regulator of cholesterol homeostasis and bile acid production, can restrict human norovirus replication in jejunal HIE monolayers. Jejunal HIE monolayers derived from two donors (J2 and J3 cell lines) were treated with increasing concentrations of 25-HC (ranging from 0.0001 µM to 5 µM) for 24 hours. Monolayers were then infected with norovirus GII.1[P33] or GII.4 Sydney[P31] and incubated for 24 hours at 37°C and 5% CO₂. Viral RNA was quantifie2025-01-01T00:00:00Z2025 Nov2026-06-06T03:08:26.433Z2026-06-06T03:06:17.378ZS-EPMC126459674111753810.1128/jvi.01109-25