Human Gut-On-A-Chip Supports Polarized Infection of Coxsackie B1 Virus In Vitro.
Analysis of enterovirus infection is difficult in animals because they express different virus receptors than humans, and static cell culture systems do not reproduce the physical complexity of the human intestinal epithelium. Here, using coxsackievirus B1 (CVB1) as a prototype enterovirus strain, we demonstrate that human enterovirus infection, replication and infectious virus production can be analyzed in vitro in a human Gut-on-a-Chip microfluidic device that supports culture of highly differentiated human villus intestinal epithelium under conditions of fluid flow and peristalsis-like motions. When CVB1 was introduced into the epithelium-lined intestinal lumen of the device, virions entered the epithelium, replicated inside the cells producing detectable cytopathic effects (CPEs), and both infectious virions and inflammatory cytokines were released in a polarized manner from the cell apex, as they could be detected in the effluent from the epithelial microchannel. When the virus was introduced via a basal route of infection (by inoculating virus into fluid flowing through a parallel lower 'vascular' channel separated from the epithelial channel by a porous membrane), significantly lower viral titers, decreased CPEs, and delayed caspase-3 activation were observed; however, cytokines continued to be secreted apically. The presence of continuous fluid flow through the epithelial lumen also resulted in production of a gradient of CPEs consistent with the flow direction. Thus, the human Gut-on-a-Chip may provide a suitable in vitro model for enteric virus infection and for investigating mechanisms of enterovirus pathogenesis.
Wyss Institute for Biologically Inspired Engineering at Harvard University, Boston, Massachusetts, United States of America.
Remi
R
Villenave
Molecular Virology Team, Division of Molecular Biology, Center for Food Safety and Applied Nutrition, U.S. Food and Drug Administration, Laurel, Maryland, United States of America.
Samantha Q
SQ
Wales
Wyss Institute for Biologically Inspired Engineering at Harvard University, Boston, Massachusetts, United States of America.
Tiama
T
Hamkins-Indik
Molecular Virology Team, Division of Molecular Biology, Center for Food Safety and Applied Nutrition, U.S. Food and Drug Administration, Laurel, Maryland, United States of America.
Efstathia
E
Papafragkou
Wyss Institute for Biologically Inspired Engineering at Harvard University, Boston, Massachusetts, United States of America.
James C
JC
Weaver
Wyss Institute for Biologically Inspired Engineering at Harvard University, Boston, Massachusetts, United States of America.
Thomas C
TC
Ferrante
Wyss Institute for Biologically Inspired Engineering at Harvard University, Boston, Massachusetts, United States of America.
Anthony
A
Bahinski
Molecular Virology Team, Division of Molecular Biology, Center for Food Safety and Applied Nutrition, U.S. Food and Drug Administration, Laurel, Maryland, United States of America.
Christopher A
CA
Elkins
Molecular Virology Team, Division of Molecular Biology, Center for Food Safety and Applied Nutrition, U.S. Food and Drug Administration, Laurel, Maryland, United States of America.
Michael
M
Kulka
Wyss Institute for Biologically Inspired Engineering at Harvard University, Boston, Massachusetts, United States of America.|Harvard John A. Paulson School of Engineering and Applied Sciences, Cambridge, Massachusetts, United States of America.|Vascular Biology Program, Boston Children's Hospital and Harvard Medical School, Boston, Massachusetts, United States of America.
Donald E
DE
Ingber
D017209:Apoptosis; D018938:Caco-2 Cells; D020169:Caspases / Q000378:metabolism; D002478:Cells, Cultured; D003384:Coxsackievirus Infections / Q000378:metabolism / Q000821:virology; D016207:Cytokines / Q000378:metabolism; D003588:Cytopathogenic Effect, Viral; D029822:Enterovirus B, Human / Q000502:physiology*; D006801:Humans; D007413:Intestinal Mucosa / Q000821:virology*; D056656:Lab-On-A-Chip Devices; D010948:Viral Plaque Assay; D014779:Virus Replication
D016207:Cytokines; D020169:Caspases