A Three-Dimensional Cell Culture Model To Study Enterovirus Infection of Polarized Intestinal Epithelial Cells

ABSTRACT Despite serving as the primary entry portal for coxsackievirus B (CVB), little is known about CVB infection of the intestinal epithelium, owing at least in part to the lack of suitable in vivo models and the inability of cultured cells to recapitulate the complexity and structure associated...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Coyne G. Drummond, Cheryl A. Nickerson, Carolyn B. Coyne
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2016
Materias:	intestinal epithelial cell enterovirus coxsackievirus B 3-D cell culture RWV bioreactor Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/6ac14801c86841b087d8d39cf9411260
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:6ac14801c86841b087d8d39cf9411260
record_format	dspace
spelling	oai:doaj.org-article:6ac14801c86841b087d8d39cf94112602021-11-15T15:21:39ZA Three-Dimensional Cell Culture Model To Study Enterovirus Infection of Polarized Intestinal Epithelial Cells10.1128/mSphere.00030-152379-5042https://doaj.org/article/6ac14801c86841b087d8d39cf94112602016-02-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mSphere.00030-15https://doaj.org/toc/2379-5042ABSTRACT Despite serving as the primary entry portal for coxsackievirus B (CVB), little is known about CVB infection of the intestinal epithelium, owing at least in part to the lack of suitable in vivo models and the inability of cultured cells to recapitulate the complexity and structure associated with the gastrointestinal (GI) tract. Here, we report on the development of a three-dimensional (3-D) organotypic cell culture model of Caco-2 cells to model CVB infection of the gastrointestinal epithelium. We show that Caco-2 cells grown in 3-D using the rotating wall vessel (RWV) bioreactor recapitulate many of the properties of the intestinal epithelium, including the formation of well-developed tight junctions, apical-basolateral polarity, brush borders, and multicellular complexity. In addition, transcriptome analyses using transcriptome sequencing (RNA-Seq) revealed the induction of a number of genes associated with intestinal epithelial differentiation and/or intestinal processes in vivo when Caco-2 cells were cultured in 3-D. Applying this model to CVB infection, we found that although the levels of intracellular virus production were similar in two-dimensional (2-D) and 3-D Caco-2 cell cultures, the release of infectious CVB was enhanced in 3-D cultures at early stages of infection. Unlike CVB, the replication of poliovirus (PV) was significantly reduced in 3-D Caco-2 cell cultures. Collectively, our studies show that Caco-2 cells grown in 3-D using the RWV bioreactor provide a cell culture model that structurally and transcriptionally represents key aspects of cells in the human GI tract and can thus be used to expand our understanding of enterovirus-host interactions in intestinal epithelial cells. IMPORTANCE Coxsackievirus B (CVB), a member of the enterovirus family of RNA viruses, is associated with meningitis, pericarditis, diabetes, dilated cardiomyopathy, and myocarditis, among other pathologies. CVB is transmitted via the fecal-oral route and encounters the epithelium lining the gastrointestinal tract early in infection. The lack of suitable in vivo and in vitro models to study CVB infection of the gastrointestinal epithelium has limited our understanding of the events that surround infection of these specialized cells. Here, we report on the development of a three-dimensional (3-D) organotypic cell culture model of human intestinal epithelial cells that better models the gastrointestinal epithelium in vivo. By applying this 3-D model, which recapitulates many aspects of the gastrointestinal epithelium in vivo, to the study of CVB infection, our work provides a new cell system to model the mechanisms by which CVB infects the intestinal epithelium, which may have a profound impact on CVB pathogenesis. Podcast: A podcast concerning this article is available.Coyne G. DrummondCheryl A. NickersonCarolyn B. CoyneAmerican Society for Microbiologyarticleintestinal epithelial cellenteroviruscoxsackievirus B3-D cell cultureRWV bioreactorMicrobiologyQR1-502ENmSphere, Vol 1, Iss 1 (2016)
institution	DOAJ
collection	DOAJ
language	EN
topic	intestinal epithelial cell enterovirus coxsackievirus B 3-D cell culture RWV bioreactor Microbiology QR1-502
spellingShingle	intestinal epithelial cell enterovirus coxsackievirus B 3-D cell culture RWV bioreactor Microbiology QR1-502 Coyne G. Drummond Cheryl A. Nickerson Carolyn B. Coyne A Three-Dimensional Cell Culture Model To Study Enterovirus Infection of Polarized Intestinal Epithelial Cells
description	ABSTRACT Despite serving as the primary entry portal for coxsackievirus B (CVB), little is known about CVB infection of the intestinal epithelium, owing at least in part to the lack of suitable in vivo models and the inability of cultured cells to recapitulate the complexity and structure associated with the gastrointestinal (GI) tract. Here, we report on the development of a three-dimensional (3-D) organotypic cell culture model of Caco-2 cells to model CVB infection of the gastrointestinal epithelium. We show that Caco-2 cells grown in 3-D using the rotating wall vessel (RWV) bioreactor recapitulate many of the properties of the intestinal epithelium, including the formation of well-developed tight junctions, apical-basolateral polarity, brush borders, and multicellular complexity. In addition, transcriptome analyses using transcriptome sequencing (RNA-Seq) revealed the induction of a number of genes associated with intestinal epithelial differentiation and/or intestinal processes in vivo when Caco-2 cells were cultured in 3-D. Applying this model to CVB infection, we found that although the levels of intracellular virus production were similar in two-dimensional (2-D) and 3-D Caco-2 cell cultures, the release of infectious CVB was enhanced in 3-D cultures at early stages of infection. Unlike CVB, the replication of poliovirus (PV) was significantly reduced in 3-D Caco-2 cell cultures. Collectively, our studies show that Caco-2 cells grown in 3-D using the RWV bioreactor provide a cell culture model that structurally and transcriptionally represents key aspects of cells in the human GI tract and can thus be used to expand our understanding of enterovirus-host interactions in intestinal epithelial cells. IMPORTANCE Coxsackievirus B (CVB), a member of the enterovirus family of RNA viruses, is associated with meningitis, pericarditis, diabetes, dilated cardiomyopathy, and myocarditis, among other pathologies. CVB is transmitted via the fecal-oral route and encounters the epithelium lining the gastrointestinal tract early in infection. The lack of suitable in vivo and in vitro models to study CVB infection of the gastrointestinal epithelium has limited our understanding of the events that surround infection of these specialized cells. Here, we report on the development of a three-dimensional (3-D) organotypic cell culture model of human intestinal epithelial cells that better models the gastrointestinal epithelium in vivo. By applying this 3-D model, which recapitulates many aspects of the gastrointestinal epithelium in vivo, to the study of CVB infection, our work provides a new cell system to model the mechanisms by which CVB infects the intestinal epithelium, which may have a profound impact on CVB pathogenesis. Podcast: A podcast concerning this article is available.
format	article
author	Coyne G. Drummond Cheryl A. Nickerson Carolyn B. Coyne
author_facet	Coyne G. Drummond Cheryl A. Nickerson Carolyn B. Coyne
author_sort	Coyne G. Drummond
title	A Three-Dimensional Cell Culture Model To Study Enterovirus Infection of Polarized Intestinal Epithelial Cells
title_short	A Three-Dimensional Cell Culture Model To Study Enterovirus Infection of Polarized Intestinal Epithelial Cells
title_full	A Three-Dimensional Cell Culture Model To Study Enterovirus Infection of Polarized Intestinal Epithelial Cells
title_fullStr	A Three-Dimensional Cell Culture Model To Study Enterovirus Infection of Polarized Intestinal Epithelial Cells
title_full_unstemmed	A Three-Dimensional Cell Culture Model To Study Enterovirus Infection of Polarized Intestinal Epithelial Cells
title_sort	three-dimensional cell culture model to study enterovirus infection of polarized intestinal epithelial cells
publisher	American Society for Microbiology
publishDate	2016
url	https://doaj.org/article/6ac14801c86841b087d8d39cf9411260
work_keys_str_mv	AT coynegdrummond athreedimensionalcellculturemodeltostudyenterovirusinfectionofpolarizedintestinalepithelialcells AT cherylanickerson athreedimensionalcellculturemodeltostudyenterovirusinfectionofpolarizedintestinalepithelialcells AT carolynbcoyne athreedimensionalcellculturemodeltostudyenterovirusinfectionofpolarizedintestinalepithelialcells AT coynegdrummond threedimensionalcellculturemodeltostudyenterovirusinfectionofpolarizedintestinalepithelialcells AT cherylanickerson threedimensionalcellculturemodeltostudyenterovirusinfectionofpolarizedintestinalepithelialcells AT carolynbcoyne threedimensionalcellculturemodeltostudyenterovirusinfectionofpolarizedintestinalepithelialcells
_version_	1718428133620711424

A Three-Dimensional Cell Culture Model To Study Enterovirus Infection of Polarized Intestinal Epithelial Cells

Ejemplares similares