Development of a novel human intestinal model to elucidate the effect of anaerobic commensals on Escherichia coli infection
The gut microbiota plays a crucial role in protecting against enteric infection. However, the underlying mechanisms are largely unknown owing to a lack of suitable experimental models. Although most gut commensals are anaerobic, intestinal epithelial cells require oxygen for survival. In addition, m...
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| Format: | Article |
| Language: | English |
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The Company of Biologists
2022-04-01
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| Series: | Disease Models & Mechanisms |
| Subjects: | |
| Online Access: | http://dmm.biologists.org/content/15/4/dmm049365 |
| _version_ | 1818470477059850240 |
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| author | Conor J. McGrath Edgaras Laveckis Andrew Bell Emmanuelle Crost Nathalie Juge Stephanie Schüller |
| author_facet | Conor J. McGrath Edgaras Laveckis Andrew Bell Emmanuelle Crost Nathalie Juge Stephanie Schüller |
| author_sort | Conor J. McGrath |
| collection | DOAJ |
| description | The gut microbiota plays a crucial role in protecting against enteric infection. However, the underlying mechanisms are largely unknown owing to a lack of suitable experimental models. Although most gut commensals are anaerobic, intestinal epithelial cells require oxygen for survival. In addition, most intestinal cell lines do not produce mucus, which provides a habitat for the microbiota. Here, we have developed a microaerobic, mucus-producing vertical diffusion chamber (VDC) model and determined the influence of Limosilactobacillus reuteri and Ruminococcus gnavus on enteropathogenic Escherichia coli (EPEC) infection. Optimization of the culture medium enabled bacterial growth in the presence of mucus-producing T84/LS174T cells. Whereas L. reuteri diminished EPEC growth and adhesion to T84/LS174T and mucus-deficient T84 epithelia, R. gnavus only demonstrated a protective effect in the presence of LS174T cells. Reduced EPEC adherence was not associated with altered type III secretion pore formation. In addition, co-culture with L. reuteri and R. gnavus dampened EPEC-induced interleukin 8 secretion. The microaerobic mucin-producing VDC system will facilitate investigations into the mechanisms underpinning colonization resistance and aid the development of microbiota-based anti-infection strategies. This article has an associated First Person interview with the first author of the paper. |
| first_indexed | 2024-04-13T21:37:37Z |
| format | Article |
| id | doaj.art-f4931e9606c84499affb7d4d15ef4427 |
| institution | Directory Open Access Journal |
| issn | 1754-8403 1754-8411 |
| language | English |
| last_indexed | 2024-04-13T21:37:37Z |
| publishDate | 2022-04-01 |
| publisher | The Company of Biologists |
| record_format | Article |
| series | Disease Models & Mechanisms |
| spelling | doaj.art-f4931e9606c84499affb7d4d15ef44272022-12-22T02:28:53ZengThe Company of BiologistsDisease Models & Mechanisms1754-84031754-84112022-04-0115410.1242/dmm.049365049365Development of a novel human intestinal model to elucidate the effect of anaerobic commensals on Escherichia coli infectionConor J. McGrath0Edgaras Laveckis1Andrew Bell2Emmanuelle Crost3Nathalie Juge4Stephanie Schüller5 Department of Clinical Medicine, Norwich Medical School, University of East Anglia, Norwich NR4 7UQ, UK Department of Clinical Medicine, Norwich Medical School, University of East Anglia, Norwich NR4 7UQ, UK Gut Microbes and Health Programme, Quadram Institute Bioscience, Gut Microbes and Health Institute Strategic Programme, Norwich NR4 7UQ, UK Gut Microbes and Health Programme, Quadram Institute Bioscience, Gut Microbes and Health Institute Strategic Programme, Norwich NR4 7UQ, UK Gut Microbes and Health Programme, Quadram Institute Bioscience, Gut Microbes and Health Institute Strategic Programme, Norwich NR4 7UQ, UK Department of Clinical Medicine, Norwich Medical School, University of East Anglia, Norwich NR4 7UQ, UK The gut microbiota plays a crucial role in protecting against enteric infection. However, the underlying mechanisms are largely unknown owing to a lack of suitable experimental models. Although most gut commensals are anaerobic, intestinal epithelial cells require oxygen for survival. In addition, most intestinal cell lines do not produce mucus, which provides a habitat for the microbiota. Here, we have developed a microaerobic, mucus-producing vertical diffusion chamber (VDC) model and determined the influence of Limosilactobacillus reuteri and Ruminococcus gnavus on enteropathogenic Escherichia coli (EPEC) infection. Optimization of the culture medium enabled bacterial growth in the presence of mucus-producing T84/LS174T cells. Whereas L. reuteri diminished EPEC growth and adhesion to T84/LS174T and mucus-deficient T84 epithelia, R. gnavus only demonstrated a protective effect in the presence of LS174T cells. Reduced EPEC adherence was not associated with altered type III secretion pore formation. In addition, co-culture with L. reuteri and R. gnavus dampened EPEC-induced interleukin 8 secretion. The microaerobic mucin-producing VDC system will facilitate investigations into the mechanisms underpinning colonization resistance and aid the development of microbiota-based anti-infection strategies. This article has an associated First Person interview with the first author of the paper.http://dmm.biologists.org/content/15/4/dmm049365epeclimosilactobacillus reuteriruminococcus gnavuscolonization resistancemicrobiotaintestinal epitheliummucusmodel system |
| spellingShingle | Conor J. McGrath Edgaras Laveckis Andrew Bell Emmanuelle Crost Nathalie Juge Stephanie Schüller Development of a novel human intestinal model to elucidate the effect of anaerobic commensals on Escherichia coli infection Disease Models & Mechanisms epec limosilactobacillus reuteri ruminococcus gnavus colonization resistance microbiota intestinal epithelium mucus model system |
| title | Development of a novel human intestinal model to elucidate the effect of anaerobic commensals on Escherichia coli infection |
| title_full | Development of a novel human intestinal model to elucidate the effect of anaerobic commensals on Escherichia coli infection |
| title_fullStr | Development of a novel human intestinal model to elucidate the effect of anaerobic commensals on Escherichia coli infection |
| title_full_unstemmed | Development of a novel human intestinal model to elucidate the effect of anaerobic commensals on Escherichia coli infection |
| title_short | Development of a novel human intestinal model to elucidate the effect of anaerobic commensals on Escherichia coli infection |
| title_sort | development of a novel human intestinal model to elucidate the effect of anaerobic commensals on escherichia coli infection |
| topic | epec limosilactobacillus reuteri ruminococcus gnavus colonization resistance microbiota intestinal epithelium mucus model system |
| url | http://dmm.biologists.org/content/15/4/dmm049365 |
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