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Characterization of a porcine intestinal epithelial cell line for in vitro studies of microbial pathogenesis in swine

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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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Authors and Affiliations

  1. Institut für Mikrobiologie und Tierseuchen, Freie Universität Berlin, Philippstr. 13, 10115, Berlin, Germany

    Peter Schierack, Marcel Nordhoff, Marion Pollmann, Jörg Jores, Sylvia Kleta, Karsten Tedin & Lothar H. Wieler

  2. Institut für Veterinär-Anatomie, Freie Universität Berlin, Koserstr. 20, 14195, Berlin, Germany

    Karl Dietrich Weyrauch

  3. Institut für Klinische Physiologie, Charitè-Universitätsmedizin Berlin, Campus Benjamin Franklin, Hindenburgdamm 30, 12200, Berlin, Germany

    Salah Amasheh

  4. Institut für Veterinär-Physiologie, Freie Universität Berlin, Oertzenweg 19b, 14163, Berlin, Germany

    Ulrike Lodemann

  5. Lehrstuhl für Molekulare Parasitologie, Humboldt-Universität Berlin, Philippstr. 13, 10115, Berlin, Germany

    Babila Tachu

  6. Department of Clinical Sciences, College of Veterinary Medicine, North Carolina State University, Raleigh, NC, 27606, USA

    Anthony Blikslager

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  1. Peter Schierack
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  2. Marcel Nordhoff
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  3. Marion Pollmann
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  8. Babila Tachu
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  9. Sylvia Kleta
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  11. Karsten Tedin
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  12. Lothar H. Wieler
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Correspondence to Peter Schierack.

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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

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