Replication of human noroviruses in stem cell–derived human enteroids

TLDR

Human noroviruses, the leading cause of gastroenteritis worldwide, have been difficult to study because no robust, reproducible in vitro cultivation system has existed. We successfully cultivated multiple HuNoV strains in stem cell–derived human intestinal enteroid monolayers, showing that bile is required, histoblood group antigen expression is essential, and that inactivation or serum neutralization blocks infection, thereby providing a system that recapitulates the intestinal epithelium and enables host‑pathogen studies and evaluation of prevention and treatment strategies.

Abstract

The major barrier to research and development of effective interventions for human noroviruses (HuNoVs) has been the lack of a robust and reproducible in vitro cultivation system. HuNoVs are the leading cause of gastroenteritis worldwide. We report the successful cultivation of multiple HuNoV strains in enterocytes in stem cell–derived, nontransformed human intestinal enteroid monolayer cultures. Bile, a critical factor of the intestinal milieu, is required for strain-dependent HuNoV replication. Lack of appropriate histoblood group antigen expression in intestinal cells restricts virus replication, and infectivity is abrogated by inactivation (e.g., irradiation, heating) and serum neutralization. This culture system recapitulates the human intestinal epithelium, permits human host-pathogen studies of previously noncultivatable pathogens, and allows the assessment of methods to prevent and treat HuNoV infections.