Replication of human noroviruses in stem cell-derived human enteroids

Authors: ETTAYEBI, KHALIL - Baylor College Of Medicine; CRAWFORD, SUE - Baylor College Of Medicine; MURAKAMI, KOSUKE - Baylor College Of Medicine; BROUGHMAN, JAMES - Baylor College Of Medicine; KARANDIKAR, UMESH - Baylor College Of Medicine; TENGE, VICTORIA - Baylor College Of Medicine; NEILL, FREDERICK - Baylor College Of Medicine; BLUTT, SARAH - Baylor College Of Medicine; ZENG, XI-LEI - Baylor College Of Medicine; QU, LIN - Baylor College Of Medicine; KOU, BAIJUN - Baylor College Of Medicine; OPEKUN, ANTONE - Baylor College Of Medicine; Burrin, Douglas - Doug; GRAHAM, DAVID - Baylor College Of Medicine; RAMANI, SASIREKHA - Baylor College Of Medicine; ATMAR, ROBERT - Baylor College Of Medicine; ESTES, MARY - Baylor College Of Medicine

Submitted to: Science
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 8/18/2016
Publication Date: 8/25/2016
Publication URL: http://handle.nal.usda.gov/10113/5430225
Citation: Ettayebi, K., Crawford, S.E., Murakami, K., Broughman, J.R., Karandikar, U., Tenge, V.R., Neill, F.H., Blutt, S.E., Zeng, X., Qu, L., Kou, B., Opekun, A.R., Burrin, D.G., Graham, D.Y., Ramani, S., Atmar, R.L., Estes, M.K. 2016. Replication of human noroviruses in stem cell-derived human enteroids. Science. doi: 10.1126/science.aaf5211.

Interpretive Summary: Human noroviruses are a leading viral cause of acute diarrhea world-wide. Noroviruses, also known as the cruise ship viruses, do not grow in laboratory cell cultures that traditionally support the growth of other viruses. Noroviruses are also species specific, in that human noroviruses only infect and cause disease in humans, do not grow in mice or other small animal models typically used for research. The aim of this study was to take a newly-developed experimental approach using tissue collected from human intestinal biopsies to grow mini-guts, also called enteroids, in cell culture that replicate what happens in the human gut. We tested the ability of human strains of norovirus to infect human enteroids and in the presence of added components normally present in gut secretions, such as enzymes and bile. We found that adding bile from humans and pigs to the cultures enabled the norovirus to infect and grow in human enteroids. Bile is important for growth of other bacterial pathogens, but this is the first time it’s been shown that bile is important for the replication of human intestinal viruses. This discovery will pave the way to develop new effective methods to prevent and treat norovirus infection and to test vaccines.

Technical 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 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.