title: Human Intestinal Enteroids: a New Model To Study Human Rotavirus Infection, Host Restriction, and Pathophysiology.
abstract: Human gastrointestinal tract research is limited by the paucity of in vitro intestinal cell models that recapitulate the cellular diversity and complex functions of human physiology and disease pathology. Human intestinal enteroid (HIE) cultures contain multiple intestinal epithelial cell types that comprise the intestinal epithelium (enterocytes and goblet, enteroendocrine, and Paneth cells) and are physiologically active based on responses to agonists. We evaluated these nontransformed, three-dimensional HIE cultures as models for pathogenic infections in the small intestine by examining whether HIEs from different regions of the small intestine from different patients are susceptible to human rotavirus (HRV) infection. Little is known about HRVs, as they generally replicate poorly in transformed cell lines, and host range restriction prevents their replication in many animal models, whereas many animal rotaviruses (ARVs) exhibit a broader host range and replicate in mice. Using HRVs, including the Rotarix RV1 vaccine strain, and ARVs, we evaluated host susceptibility, virus production, and cellular responses of HIEs. HRVs infect at higher rates and grow to higher titers than do ARVs. HRVs infect differentiated enterocytes and enteroendocrine cells, and viroplasms and lipid droplets are induced. Heterogeneity in replication was seen in HIEs from different patients. HRV infection and RV enterotoxin treatment of HIEs caused physiological lumenal expansion detected by time-lapse microscopy, recapitulating one of the hallmarks of rotavirus-induced diarrhea. These results demonstrate that HIEs are a novel pathophysiological model that will allow the study of HRV biology, including host restriction, cell type restriction, and virus-induced fluid secretion.
  
  
  OBJECTIVE
  Our research establishes HIEs as nontransformed cell culture models to understand human intestinal physiology and pathophysiology and the epithelial response, including host restriction of gastrointestinal infections such as HRV infection. HRVs remain a major worldwide cause of diarrhea-associated morbidity and mortality in children ≤5 years of age. Current in vitro models of rotavirus infection rely primarily on the use of animal rotaviruses because HRV growth is limited in most transformed cell lines and animal models. We demonstrate that HIEs are novel, cellularly diverse, and physiologically relevant epithelial cell cultures that recapitulate in vivo properties of HRV infection. HIEs will allow the study of HRV biology, including human host-pathogen and live, attenuated vaccine interactions; host and cell type restriction; virus-induced fluid secretion; cell-cell communication within the epithelium; and the epithelial response to infection in cultures from genetically diverse individuals. Finally, drug therapies to prevent/treat diarrheal disease can be tested in these physiologically active cultures.
authors.affiliation: Department of Molecular Virology and Microbiology, Baylor College of Medicine, Houston, Texas, USA.
authors.forename: Kapil
authors.initials: K
authors.lastname: Saxena
authors.affiliation: Department of Molecular Virology and Microbiology, Baylor College of Medicine, Houston, Texas, USA.
authors.forename: Sarah E
authors.initials: SE
authors.lastname: Blutt
authors.affiliation: Department of Molecular Virology and Microbiology, Baylor College of Medicine, Houston, Texas, USA.
authors.forename: Khalil
authors.initials: K
authors.lastname: Ettayebi
authors.affiliation: Department of Molecular Virology and Microbiology, Baylor College of Medicine, Houston, Texas, USA.
authors.forename: Xi-Lei
authors.initials: XL
authors.lastname: Zeng
authors.affiliation: Department of Molecular Virology and Microbiology, Baylor College of Medicine, Houston, Texas, USA.
authors.forename: James R
authors.initials: JR
authors.lastname: Broughman
authors.affiliation: Department of Molecular Virology and Microbiology, Baylor College of Medicine, Houston, Texas, USA.
authors.forename: Sue E
authors.initials: SE
authors.lastname: Crawford
authors.affiliation: Department of Molecular Virology and Microbiology, Baylor College of Medicine, Houston, Texas, USA.
authors.forename: Umesh C
authors.initials: UC
authors.lastname: Karandikar
authors.affiliation: Department of Molecular Virology and Microbiology, Baylor College of Medicine, Houston, Texas, USA.
authors.forename: Narayan P
authors.initials: NP
authors.lastname: Sastri
authors.affiliation: Department of Molecular Virology and Microbiology, Baylor College of Medicine, Houston, Texas, USA.
authors.forename: Margaret E
authors.initials: ME
authors.lastname: Conner
authors.affiliation: Department of Medicine, Baylor College of Medicine, Houston, Texas, USA.
authors.forename: Antone R
authors.initials: AR
authors.lastname: Opekun
authors.affiliation: Department of Molecular Virology and Microbiology, Baylor College of Medicine, Houston, Texas, USA Department of Medicine, Baylor College of Medicine, Houston, Texas, USA.
authors.forename: David Y
authors.initials: DY
authors.lastname: Graham
authors.affiliation: Department of Medicine, Baylor College of Medicine, Houston, Texas, USA.
authors.forename: Waqar
authors.initials: W
authors.lastname: Qureshi
authors.affiliation: Department of Surgery, Minimally Invasive Bariatric and General Division, Houston Methodist Hospital, Houston, Texas, USA.
authors.forename: Vadim
authors.initials: V
authors.lastname: Sherman
authors.affiliation: Department of Medicine, Gastroenterology Division, Johns Hopkins University School of Medicine, Baltimore, Maryland, USA.
authors.forename: Jennifer
authors.initials: J
authors.lastname: Foulke-Abel
authors.affiliation: Department of Medicine, Gastroenterology Division, Johns Hopkins University School of Medicine, Baltimore, Maryland, USA.
authors.forename: Julie
authors.initials: J
authors.lastname: In
authors.affiliation: Department of Medicine, Gastroenterology Division, Johns Hopkins University School of Medicine, Baltimore, Maryland, USA.
authors.forename: Olga
authors.initials: O
authors.lastname: Kovbasnjuk
authors.affiliation: Department of Medicine, Gastroenterology Division, Johns Hopkins University School of Medicine, Baltimore, Maryland, USA.
authors.forename: Nicholas C
authors.initials: NC
authors.lastname: Zachos
authors.affiliation: Department of Medicine, Gastroenterology Division, Johns Hopkins University School of Medicine, Baltimore, Maryland, USA.
authors.forename: Mark
authors.initials: M
authors.lastname: Donowitz
authors.affiliation: Department of Molecular Virology and Microbiology, Baylor College of Medicine, Houston, Texas, USA Department of Medicine, Baylor College of Medicine, Houston, Texas, USA mestes@bcm.tmc.edu.
authors.forename: Mary K
authors.initials: MK
authors.lastname: Estes
mesh_terms: D006801:Humans; D007421:Intestine, Small / Q000502:physiology / Q000821:virology*; D008962:Models, Theoretical; D009924:Organ Culture Techniques / Q000379:methods*; D012401:Rotavirus / Q000502:physiology*; D012400:Rotavirus Infections / Q000473:pathology* / Q000821:virology*; D014779:Virus Replication