TY - JOUR
T1 - Evaluating shigella flexneri pathogenesis in the human enteroid model
AU - Ranganathan, Sridevi
AU - Doucet, Michele
AU - Grassel, Christen L.
AU - Delaine-Elias, Bre Onna
AU - Zachos, Nicholas C.
AU - Barry, Eileen M.
N1 - Funding Information:
This work was supported by NIH grant P01AI125181 and NIH grant U19AI109776. We also acknowledge the Confocal Microscopy Core at the University of Maryland, Baltimore, and the Integrated Physiology Core of the Hopkins Conte Digestive Disease Basic and Translational Research Core Center (NIH grant P30DK089502).
Publisher Copyright:
Copyright © 2019 Ranganathan et al. This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license.
PY - 2019/4/1
Y1 - 2019/4/1
N2 - The enteric pathogen Shigella is one of the leading causes of moderate-to-severe diarrhea and death in young children in developing countries. Transformed cell lines and animal models have been widely used to study Shigella pathogenesis. In addition to altered physiology, transformed cell lines are composed of a single cell type that does not sufficiently represent the complex multicellular environment of the human colon. Most available animal models do not accurately mimic human disease. The human intestinal enteroid model, derived from LGR5 stem cell-containing intestinal crypts from healthy subjects, represents a technological leap in human gastrointestinal system modeling and provides a more physiologically relevant system that includes multiple cell types and features of the human intestine. We established the utility of this model for studying basic aspects of Shigella pathogenesis and host responses. In this study, we show that Shigella flexneri is capable of infecting and replicating intracellularly in human enteroids derived from different segments of the intestine. Apical invasion by S. flexneri is very limited but increases 10-fold when enteroids are differentiated to include M cells. Invasion via the basolateral surface was at least 2-log10 units more efficient than apical infection. Increased secretion of interleukin-8 and higher expression levels of the mucin glycoprotein Muc2 were observed in the enteroids following S. flexneri infection. The human enteroid model promises to bridge some of the gaps between traditional cell culture, animal models, and human infection.
AB - The enteric pathogen Shigella is one of the leading causes of moderate-to-severe diarrhea and death in young children in developing countries. Transformed cell lines and animal models have been widely used to study Shigella pathogenesis. In addition to altered physiology, transformed cell lines are composed of a single cell type that does not sufficiently represent the complex multicellular environment of the human colon. Most available animal models do not accurately mimic human disease. The human intestinal enteroid model, derived from LGR5 stem cell-containing intestinal crypts from healthy subjects, represents a technological leap in human gastrointestinal system modeling and provides a more physiologically relevant system that includes multiple cell types and features of the human intestine. We established the utility of this model for studying basic aspects of Shigella pathogenesis and host responses. In this study, we show that Shigella flexneri is capable of infecting and replicating intracellularly in human enteroids derived from different segments of the intestine. Apical invasion by S. flexneri is very limited but increases 10-fold when enteroids are differentiated to include M cells. Invasion via the basolateral surface was at least 2-log10 units more efficient than apical infection. Increased secretion of interleukin-8 and higher expression levels of the mucin glycoprotein Muc2 were observed in the enteroids following S. flexneri infection. The human enteroid model promises to bridge some of the gaps between traditional cell culture, animal models, and human infection.
KW - Colonoid
KW - Human intestinal enteroids
KW - M cells
KW - S. flexneri pathogenesis
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U2 - 10.1128/IAI.00740-18
DO - 10.1128/IAI.00740-18
M3 - Article
C2 - 30642900
AN - SCOPUS:85063712974
SN - 0019-9567
VL - 87
JO - Infection and immunity
JF - Infection and immunity
IS - 4
M1 - e00740
ER -