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...
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Taylor & Francis Group
2023-12-01
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| Series: | Emerging Microbes and Infections |
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| Online Access: | https://www.tandfonline.com/doi/10.1080/22221751.2023.2239937 |
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| author | Miaomiao Yan Ang Su Suvarin Pavasutthipaisit Rebecca Spriewald Guntram A. Graßl Andreas Beineke Doris Hoeltig Georg Herrler Paul Becher |
| author_facet | Miaomiao Yan Ang Su Suvarin Pavasutthipaisit Rebecca Spriewald Guntram A. Graßl Andreas Beineke Doris Hoeltig Georg Herrler Paul Becher |
| author_sort | Miaomiao Yan |
| collection | DOAJ |
| 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 | Article |
| id | doaj-art-84ed079bf3b54aca89c03247cfe1aa97 |
| institution | OA Journals |
| issn | 2222-1751 |
| language | English |
| publishDate | 2023-12-01 |
| publisher | Taylor & Francis Group |
| record_format | Article |
| series | Emerging Microbes and Infections |
| spelling | doaj-art-84ed079bf3b54aca89c03247cfe1aa972025-08-20T02:37:28ZengTaylor & Francis GroupEmerging Microbes and Infections2222-17512023-12-0112210.1080/22221751.2023.2239937Infection of porcine enteroids and 2D differentiated intestinal epithelial cells with rotavirus A to study cell tropism and polarized immune responseMiaomiao Yan0Ang Su1Suvarin Pavasutthipaisit2Rebecca Spriewald3Guntram A. Graßl4Andreas Beineke5Doris Hoeltig6Georg Herrler7Paul Becher8Institute of Virology, University of Veterinary Medicine Hannover, Hannover, GermanyInstitute of Virology, University of Veterinary Medicine Hannover, Hannover, GermanyInstitute of Pathology, University of Veterinary Medicine Hannover, Hannover, GermanyInstitute of Microbiology, University of Veterinary Medicine Hannover, Hannover, GermanyInstitute of Medical Microbiology and Hospital Epidemiology, Hannover Medical School and German Center for Infection Research (DZIF), Hannover, GermanyInstitute of Pathology, University of Veterinary Medicine Hannover, Hannover, GermanyClinic for Swine and Small Ruminants, Forensic Medicine and Ambulatory Service, University of Veterinary Medicine Hannover, Hannover, GermanyInstitute of Virology, University of Veterinary Medicine Hannover, Hannover, GermanyInstitute of Virology, University of Veterinary Medicine Hannover, Hannover, GermanyIntestinal 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.https://www.tandfonline.com/doi/10.1080/22221751.2023.2239937Rotavirus Aenteroidsenterocytesgoblet cellsPaneth cellsinnate immunity |
| spellingShingle | Miaomiao Yan Ang Su Suvarin Pavasutthipaisit Rebecca Spriewald Guntram A. Graßl Andreas Beineke Doris Hoeltig Georg Herrler Paul Becher Infection of porcine enteroids and 2D differentiated intestinal epithelial cells with rotavirus A to study cell tropism and polarized immune response Emerging Microbes and Infections Rotavirus A enteroids enterocytes goblet cells Paneth cells innate immunity |
| 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 | Rotavirus A enteroids enterocytes goblet cells Paneth cells innate immunity |
| url | https://www.tandfonline.com/doi/10.1080/22221751.2023.2239937 |
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