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 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.
Schierack
Peter
P
Institut für Mikrobiologie und Tierseuchen, Freie Universität Berlin, Berlin, Germany. schierack.peter@vetmed.fu-berlin.de
Nordhoff
Marcel
M
Pollmann
Marion
M
Weyrauch
Karl Dietrich
KD
Amasheh
Salah
S
Lodemann
Ulrike
U
Jores
Jörg
J
Tachu
Babila
B
Kleta
Sylvia
S
Blikslager
Anthony
A
Tedin
Karsten
K
Wieler
Lothar H
LH
D000818:Animals; D001419:Bacteria / Q000472:pathogenicity; D001424:Bacterial Infections / Q000209:etiology / Q000382:microbiology / Q000662:veterinary; D001483:Base Sequence; D002460:Cell Line; D016207:Cytokines / Q000235:genetics; D017931:DNA Primers / Q000235:genetics; D004847:Epithelial Cells / Q000382:microbiology; D004927:Escherichia coli Infections / Q000209:etiology / Q000382:microbiology / Q000662:veterinary; D066298:In Vitro Techniques; D007422:Intestines / Q000166:cytology / Q000382:microbiology*; D008854:Microscopy, Electron; D008856:Microscopy, Fluorescence; D008954:Models, Biological; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012481:Salmonella Infections, Animal / Q000209:etiology / Q000382:microbiology; D013552:Swine / Q000382:microbiology*; D013553:Swine Diseases / Q000209:etiology / Q000382:microbiology
D016207:Cytokines; D017931:DNA Primers