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. 2010:2010:469583.
doi: 10.1155/2010/469583. Epub 2010 Dec 29.

Preliminary Characterization of the Transcriptional Response of the Porcine Intestinal Cell Line IPEC-J2 to Enterotoxigenic Escherichia coli, Escherichia coli, and E. coli Lipopolysaccharide

Marisa M Geens  1 , Theo A Niewold
Affiliations

Preliminary Characterization of the Transcriptional Response of the Porcine Intestinal Cell Line IPEC-J2 to Enterotoxigenic Escherichia coli, Escherichia coli, and E. coli Lipopolysaccharide

Marisa M Geens et al. Comp Funct Genomics. 2010.

Abstract

IPEC-J2, a promising in vitro model system, is not well characterized especially on the transcriptional level, in contrast to human counterparts. The aim of this study was to characterize the gene expression in IPEC-J2 cells when coincubated with enterotoxigenic Escherichia coli (ETEC), nonpathogenic E. coli, and E. coli endotoxin. Apical infection of polarized IPEC-J2 monolayers caused a time-dependent decrease in transepithelial electrical resistance (TEER). Microarray analysis showed up-regulation of interleukins when IPEC-J2 were cocultured with E. coli strains this has so far never been measured in this cell line. Highest IL8 expression was found with the ETEC strain possessing the F4 fimbrium, suggesting IPEC-J2 cells to be F4 receptor positive, confirmed in a brush border membrane adhesion assay. It is concluded that the innate immune responses to pathogens and LPS makes the IPEC-J2 cell line a suitable model for research on intestinal host pathogen interaction.

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Figures

Figure 1
Figure 1
Linear regression of qRT-PCR data versus microarray data of IL8, IRG6, CYP1A1 and FABP5. The goodness of fit (r 2) and P-value are given.
Figure 2
Figure 2
qRT-PCR analysis of IL8, IRG6, CYP1A1 and FABP5, in response to the four treatments at 4 h. LPS (1), CVI-444 (2), CVI-1000 (3) and CVI-1048 (4) (mean with S.D. of n = 5-6). Asterisks indicate significant differences between treatments. *.05 < P < .01, **.01 < P < .005, ***P < .005.
Figure 3
Figure 3
Intracellular concentration of I-FABP (mean ± S.D. of n = 6–8) in the three different cell lines during 14 days of culture (open circle: IPEC-J2 cells, closed triangle: COS-7 cells and closed rectangle: T84 cells).
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