Abstract
In vitro studies on the pathogenesis in swine have been hampered by the lack of relevant porcine cell lines. Since many bacterial infections are swine-specific, studies on pathogenic mechanisms require appropriate cell lines of porcine origin. We have characterized the permanent porcine intestinal epithelial cell line, IPEC-J2, using a variety of methods in order to assess the usefulness of this cell line as an in vitro infection model. Electron microscopic analyses and histochemical staining revealed the cells to be enterocyte-like with microvilli, tight junctions and glycocalyx-bound mucin. The functional integrity of monolayers was determined by transepithelial electrical resistance (TEER) measurements. Both commensal bacteria and important bacterial pathogens were chosen for study based on their principally different infection mechanisms: obligate extracellular Escherichia coli, facultative intracellular Salmonella and obligate intracellular Chlamydia. We determined the colonization and proliferation of the bacteria on and within the host cells and monitored the host cell response. We verified the expression of mRNAs encoding the cytokines IL-1α, −6, −7, −8, −18, TNF-α and GM-CSF, but not TGF-β or MCP-1. IL-8 protein expression was enhanced by Salmonella invasion. We conclude that the IPEC-J2 cell line provides a relevant in vitro model system for porcine intestinal pathogen–host cell interactions.
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Abbreviations
- AE:
-
Attaching and effacing
- CLSM:
-
Confocal laser scanning microscopy
- DAPI:
-
4′,6-Diamidino-2-phenylindole
- DMEM:
-
Dulbecco’s modified eagle medium
- EGF:
-
Epidermal growth factor
- EPEC:
-
Enteropathogenic E. coli
- FCS:
-
Fetal calf serum
- FITC:
-
Fluoresceinisothiocyanate
- GFP:
-
Green fluorescent protein
- GM-CSF:
-
Granulocyte macrophage colony stimulating factor
- IFN:
-
Interferon
- ITS:
-
Insulin transferrin selenium
- MOI:
-
Multiplicity of infection
- PAS:
-
Periodic acid-Schiff reaction
- PBS:
-
Phosphate buffered saline
- RT:
-
Reverse transcriptase
- TEER:
-
Transepithelial electrical resistance
- TNF:
-
Tumor necrosis factor
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Acknowledgements
We thank H. Steinrück (Bundesinstitut für Risikobewertung, Berlin, Germany) for serotyping of E. coli, B. Seeger (Neurowissenschaftliches Forschungszentrum, Berlin, Germany) for assistance with the proliferation assays and L. Scharek (Institut für Immunologie und Molekularbiologie, Berlin, Germany) for fluorescence activated cell scanning. This work was supported by grant FOR 438/1-1 from the Deutsche Forschungsgemeinschaft. M. Pollmann was supported by the Studienstiftung des deutschen Volkes, S. Kleta by NaFöG Berlin.
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Schierack, P., Nordhoff, M., Pollmann, M. et al. Characterization of a porcine intestinal epithelial cell line for in vitro studies of microbial pathogenesis in swine. Histochem Cell Biol 125, 293–305 (2006). https://doi.org/10.1007/s00418-005-0067-z
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DOI: https://doi.org/10.1007/s00418-005-0067-z