TY - JOUR
T1 - Human intestinal enteroids
T2 - A new model to study human rotavirus infection, host restriction, and pathophysiology
AU - Saxena, Kapil
AU - Blutt, Sarah E.
AU - Ettayebi, Khalil
AU - Zeng, Xi Lei
AU - Broughman, James R.
AU - Crawford, Sue E.
AU - Karandikar, Umesh C.
AU - Sastri, Narayan P.
AU - Conner, Margaret E.
AU - Opekun, Antone R.
AU - Graham, David Y.
AU - Qureshi, Waqar
AU - Sherman, Vadim
AU - Foulke-Abel, Jennifer
AU - In, Julie
AU - Kovbasnjuk, Olga
AU - Zachos, Nicholas C.
AU - Donowitz, Mark
AU - Estes, Mary K.
N1 - Funding Information:
HHS | NIH | National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK) provided funding to Mary Estes. HHS| NIH | National Institute of Allergy and Infectious Diseases (NIAID) provided funding to James R. Broughman, Sue Ellen Crawford, Narayan P. Sastri, and Mary Estes under grant number RO1-AI080656.HHS| NIH | National Institute of Allergy and Infectious Diseases (NIAID) provided funding to Sarah Blutt, Xi-Lei Zeng, Kapil Saxena, Sue Crawford, and Mary Estes under grant number U19-AI11497. HHS | NIH | National Center for Advancing Translational Sciences (NCATS) provided funding to Sarah Blutt, Khalil Ettayebi, Xi-Lei Zeng, Umesh Karandikar, Jennifer Foulke-Abel, Julie In, Olga Kovbasnjuk, Nicholas C. Zachos, Mark Donowitz, and Mary K. Estes under grant number U18-TR000552. Howard Hughes Medical Institute (HHMI) provided funding to Kapil Saxena under grant number 570076890. The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.
Publisher Copyright:
© 2015, American Society for Microbiology.
PY - 2016
Y1 - 2016
N2 - 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.
AB - 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.
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U2 - 10.1128/JVI.01930-15
DO - 10.1128/JVI.01930-15
M3 - Article
C2 - 26446608
AN - SCOPUS:84953923549
SN - 0022-538X
VL - 90
SP - 43
EP - 56
JO - Journal of virology
JF - Journal of virology
IS - 1
ER -