Cargando…
Infection of porcine enteroids and 2D differentiated intestinal epithelial cells with rotavirus A to study cell tropism and polarized immune response
Intestinal epithelial cell interactions with enteric pathogens have been incompletely elucidated owing to the lack of model systems that recapitulate the cellular diversity, architecture and functionality of the intestine. To analyze rotavirus (RV) infection and the subsequent innate immune response...
Autores principales: | , , , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Taylor & Francis
2023
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10399479/ https://www.ncbi.nlm.nih.gov/pubmed/37483148 http://dx.doi.org/10.1080/22221751.2023.2239937 |
_version_ | 1785084255001378816 |
---|---|
author | Yan, Miaomiao Su, Ang Pavasutthipaisit, Suvarin Spriewald, Rebecca Graßl, Guntram A. Beineke, Andreas Hoeltig, Doris Herrler, Georg Becher, Paul |
author_facet | Yan, Miaomiao Su, Ang Pavasutthipaisit, Suvarin Spriewald, Rebecca Graßl, Guntram A. Beineke, Andreas Hoeltig, Doris Herrler, Georg Becher, Paul |
author_sort | Yan, Miaomiao |
collection | PubMed |
description | Intestinal epithelial cell interactions with enteric pathogens have been incompletely elucidated owing to the lack of model systems that recapitulate the cellular diversity, architecture and functionality of the intestine. To analyze rotavirus (RV) infection and the subsequent innate immune response, we established cultures of differentiated porcine intestinal epithelial cells in three different variations: basolateral-out enteroids, apical-out enteroids and two-dimensional (2D) filter-grown intestinal epithelial cells. Application of specific antibodies for fluorescent staining indicated that enteroids and enteroid-derived cell cultures contain multiple intestinal epithelial cell types. Infection studies indicated that both apical-out enteroids and 2D intestinal epithelial cells are susceptible to porcine RV infection. However, 2D intestinal epithelial cells are more useful for a detailed characterization and comparison of apical and basolateral infection than apical-out enteroids. Virus-induced apoptosis was observed in apical-out enteroids at 24 h post infection but not at earlier time points after infection. RV infected not only enterocytes but also goblet cells and Paneth cells in apical-out enteroids and 2D intestinal epithelial cells. Interestingly, despite the lack of significant differences in the efficiency of infection after apical and basolateral infection of 2D intestinal epithelial cells, stronger innate immune and inflammatory responses were observed after basolateral infection as compared to infection via the apical route. Therefore, apical-out enteroids and 2D intestinal epithelial cells provide useful primary cell culture models that can be extended to analyze invasion and replication strategies of agents implicated in enteric diseases or to study immune and inflammatory responses of the host induced by enteric pathogens. |
format | Online Article Text |
id | pubmed-10399479 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | Taylor & Francis |
record_format | MEDLINE/PubMed |
spelling | pubmed-103994792023-08-04 Infection of porcine enteroids and 2D differentiated intestinal epithelial cells with rotavirus A to study cell tropism and polarized immune response Yan, Miaomiao Su, Ang Pavasutthipaisit, Suvarin Spriewald, Rebecca Graßl, Guntram A. Beineke, Andreas Hoeltig, Doris Herrler, Georg Becher, Paul Emerg Microbes Infect Research Article Intestinal epithelial cell interactions with enteric pathogens have been incompletely elucidated owing to the lack of model systems that recapitulate the cellular diversity, architecture and functionality of the intestine. To analyze rotavirus (RV) infection and the subsequent innate immune response, we established cultures of differentiated porcine intestinal epithelial cells in three different variations: basolateral-out enteroids, apical-out enteroids and two-dimensional (2D) filter-grown intestinal epithelial cells. Application of specific antibodies for fluorescent staining indicated that enteroids and enteroid-derived cell cultures contain multiple intestinal epithelial cell types. Infection studies indicated that both apical-out enteroids and 2D intestinal epithelial cells are susceptible to porcine RV infection. However, 2D intestinal epithelial cells are more useful for a detailed characterization and comparison of apical and basolateral infection than apical-out enteroids. Virus-induced apoptosis was observed in apical-out enteroids at 24 h post infection but not at earlier time points after infection. RV infected not only enterocytes but also goblet cells and Paneth cells in apical-out enteroids and 2D intestinal epithelial cells. Interestingly, despite the lack of significant differences in the efficiency of infection after apical and basolateral infection of 2D intestinal epithelial cells, stronger innate immune and inflammatory responses were observed after basolateral infection as compared to infection via the apical route. Therefore, apical-out enteroids and 2D intestinal epithelial cells provide useful primary cell culture models that can be extended to analyze invasion and replication strategies of agents implicated in enteric diseases or to study immune and inflammatory responses of the host induced by enteric pathogens. Taylor & Francis 2023-08-02 /pmc/articles/PMC10399479/ /pubmed/37483148 http://dx.doi.org/10.1080/22221751.2023.2239937 Text en © 2023 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group, on behalf of Shanghai Shangyixun Cultural Communication Co., Ltd https://creativecommons.org/licenses/by-nc/4.0/This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial License (http://creativecommons.org/licenses/by-nc/4.0/ (https://creativecommons.org/licenses/by-nc/4.0/) ), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited. The terms on which this article has been published allow the posting of the Accepted Manuscript in a repository by the author(s) or with their consent. |
spellingShingle | Research Article Yan, Miaomiao Su, Ang Pavasutthipaisit, Suvarin Spriewald, Rebecca Graßl, Guntram A. Beineke, Andreas Hoeltig, Doris Herrler, Georg Becher, Paul Infection of porcine enteroids and 2D differentiated intestinal epithelial cells with rotavirus A to study cell tropism and polarized immune response |
title | Infection of porcine enteroids and 2D differentiated intestinal epithelial cells with rotavirus A to study cell tropism and polarized immune response |
title_full | Infection of porcine enteroids and 2D differentiated intestinal epithelial cells with rotavirus A to study cell tropism and polarized immune response |
title_fullStr | Infection of porcine enteroids and 2D differentiated intestinal epithelial cells with rotavirus A to study cell tropism and polarized immune response |
title_full_unstemmed | Infection of porcine enteroids and 2D differentiated intestinal epithelial cells with rotavirus A to study cell tropism and polarized immune response |
title_short | Infection of porcine enteroids and 2D differentiated intestinal epithelial cells with rotavirus A to study cell tropism and polarized immune response |
title_sort | infection of porcine enteroids and 2d differentiated intestinal epithelial cells with rotavirus a to study cell tropism and polarized immune response |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10399479/ https://www.ncbi.nlm.nih.gov/pubmed/37483148 http://dx.doi.org/10.1080/22221751.2023.2239937 |
work_keys_str_mv | AT yanmiaomiao infectionofporcineenteroidsand2ddifferentiatedintestinalepithelialcellswithrotavirusatostudycelltropismandpolarizedimmuneresponse AT suang infectionofporcineenteroidsand2ddifferentiatedintestinalepithelialcellswithrotavirusatostudycelltropismandpolarizedimmuneresponse AT pavasutthipaisitsuvarin infectionofporcineenteroidsand2ddifferentiatedintestinalepithelialcellswithrotavirusatostudycelltropismandpolarizedimmuneresponse AT spriewaldrebecca infectionofporcineenteroidsand2ddifferentiatedintestinalepithelialcellswithrotavirusatostudycelltropismandpolarizedimmuneresponse AT graßlguntrama infectionofporcineenteroidsand2ddifferentiatedintestinalepithelialcellswithrotavirusatostudycelltropismandpolarizedimmuneresponse AT beinekeandreas infectionofporcineenteroidsand2ddifferentiatedintestinalepithelialcellswithrotavirusatostudycelltropismandpolarizedimmuneresponse AT hoeltigdoris infectionofporcineenteroidsand2ddifferentiatedintestinalepithelialcellswithrotavirusatostudycelltropismandpolarizedimmuneresponse AT herrlergeorg infectionofporcineenteroidsand2ddifferentiatedintestinalepithelialcellswithrotavirusatostudycelltropismandpolarizedimmuneresponse AT becherpaul infectionofporcineenteroidsand2ddifferentiatedintestinalepithelialcellswithrotavirusatostudycelltropismandpolarizedimmuneresponse |