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Characterization of a porcine intestinal epithelial cell line for in vitro studies of microbial pathogenesis in swine
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 e...
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Published in: | Histochemistry and cell biology 2006-03, Vol.125 (3), p.293-305 |
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creator | Schierack, Peter Nordhoff, Marcel Pollmann, Marion Weyrauch, Karl Dietrich Amasheh, Salah Lodemann, Ulrike Jores, Jörg Tachu, Babila Kleta, Sylvia Blikslager, Anthony Tedin, Karsten Wieler, Lothar H |
description | 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-1alpha, -6, -7, -8, -18, TNF-alpha and GM-CSF, but not TGF-beta 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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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-1alpha, -6, -7, -8, -18, TNF-alpha and GM-CSF, but not TGF-beta 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.</description><identifier>ISSN: 0948-6143</identifier><identifier>EISSN: 1432-119X</identifier><identifier>DOI: 10.1007/s00418-005-0067-z</identifier><identifier>PMID: 16215741</identifier><language>eng</language><publisher>Germany: Springer Nature B.V</publisher><subject>Animals ; Bacteria - pathogenicity ; Bacterial Infections - etiology ; Bacterial Infections - microbiology ; Bacterial Infections - veterinary ; Base Sequence ; Cell Line ; Cytokines - genetics ; DNA Primers - genetics ; Epithelial Cells - microbiology ; Escherichia coli Infections - etiology ; Escherichia coli Infections - microbiology ; Escherichia coli Infections - veterinary ; In Vitro Techniques ; Intestines - cytology ; Intestines - microbiology ; Microscopy, Electron ; Microscopy, Fluorescence ; Models, Biological ; Reverse Transcriptase Polymerase Chain Reaction ; Salmonella Infections, Animal - etiology ; Salmonella Infections, Animal - microbiology ; Swine - microbiology ; Swine Diseases - etiology ; Swine Diseases - microbiology</subject><ispartof>Histochemistry and cell biology, 2006-03, Vol.125 (3), p.293-305</ispartof><rights>Springer-Verlag 2006</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c392t-fcdacf6bb5646c84b6959b3a62a1bc409ecf45acda0c0e077d6cfc7a9310be963</citedby><cites>FETCH-LOGICAL-c392t-fcdacf6bb5646c84b6959b3a62a1bc409ecf45acda0c0e077d6cfc7a9310be963</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,780,784,27924,27925</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/16215741$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Schierack, Peter</creatorcontrib><creatorcontrib>Nordhoff, Marcel</creatorcontrib><creatorcontrib>Pollmann, Marion</creatorcontrib><creatorcontrib>Weyrauch, Karl Dietrich</creatorcontrib><creatorcontrib>Amasheh, Salah</creatorcontrib><creatorcontrib>Lodemann, Ulrike</creatorcontrib><creatorcontrib>Jores, Jörg</creatorcontrib><creatorcontrib>Tachu, Babila</creatorcontrib><creatorcontrib>Kleta, Sylvia</creatorcontrib><creatorcontrib>Blikslager, Anthony</creatorcontrib><creatorcontrib>Tedin, Karsten</creatorcontrib><creatorcontrib>Wieler, Lothar H</creatorcontrib><title>Characterization of a porcine intestinal epithelial cell line for in vitro studies of microbial pathogenesis in swine</title><title>Histochemistry and cell biology</title><addtitle>Histochem Cell Biol</addtitle><description>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-1alpha, -6, -7, -8, -18, TNF-alpha and GM-CSF, but not TGF-beta 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.</description><subject>Animals</subject><subject>Bacteria - pathogenicity</subject><subject>Bacterial Infections - etiology</subject><subject>Bacterial Infections - microbiology</subject><subject>Bacterial Infections - veterinary</subject><subject>Base Sequence</subject><subject>Cell Line</subject><subject>Cytokines - genetics</subject><subject>DNA Primers - genetics</subject><subject>Epithelial Cells - microbiology</subject><subject>Escherichia coli Infections - etiology</subject><subject>Escherichia coli Infections - microbiology</subject><subject>Escherichia coli Infections - veterinary</subject><subject>In Vitro Techniques</subject><subject>Intestines - cytology</subject><subject>Intestines - microbiology</subject><subject>Microscopy, Electron</subject><subject>Microscopy, Fluorescence</subject><subject>Models, Biological</subject><subject>Reverse Transcriptase Polymerase Chain Reaction</subject><subject>Salmonella Infections, Animal - 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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-1alpha, -6, -7, -8, -18, TNF-alpha and GM-CSF, but not TGF-beta 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.</abstract><cop>Germany</cop><pub>Springer Nature B.V</pub><pmid>16215741</pmid><doi>10.1007/s00418-005-0067-z</doi><tpages>13</tpages></addata></record> |
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subjects | Animals Bacteria - pathogenicity Bacterial Infections - etiology Bacterial Infections - microbiology Bacterial Infections - veterinary Base Sequence Cell Line Cytokines - genetics DNA Primers - genetics Epithelial Cells - microbiology Escherichia coli Infections - etiology Escherichia coli Infections - microbiology Escherichia coli Infections - veterinary In Vitro Techniques Intestines - cytology Intestines - microbiology Microscopy, Electron Microscopy, Fluorescence Models, Biological Reverse Transcriptase Polymerase Chain Reaction Salmonella Infections, Animal - etiology Salmonella Infections, Animal - microbiology Swine - microbiology Swine Diseases - etiology Swine Diseases - microbiology |
title | Characterization of a porcine intestinal epithelial cell line for in vitro studies of microbial pathogenesis in swine |
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