Abstract
The IPEC-J2 cell line was studied as a simple model for investigating responses of the newborn intestinal epithelium to diets. Especially, the small intestine of immature newborns is sensitive to diet-induced inflammation. We investigated gene expression of epithelial- and immune response-related genes in IPEC-J2 cells stimulated for 2 h with milk formula (CELL-FORM), colostrum (CELL-COLOS), or growth medium (CELL-CONTR) and in distal small intestinal tissue samples from preterm pigs fed milk formula (PIG-FORM) or colostrum (PIG-COLOS). High throughput quantitative PCR analysis of 48 genes revealed the expression of 22 genes in IPEC-J2 cells and 31 genes in intestinal samples. Principal component analysis (PCA) discriminated the gene expression profile of IPEC-J2 cells from that of intestinal samples. The expression profile of intestinal tissue was separated by PCA into 2 groups according to diet, whereas no diet-dependent grouping was seen for IPEC-J2 cells. Expression differences between PIG-FORM and PIG-COLOS were found for DEFB1, CXCL10, IL1RN, and ALPI, while IL8 was upregulated in CELL-FORM compared with CELL-CONTR. These differences, between IPEC-J2 cells and intestinal tissue from preterm pigs, both used as models for the newborn intestine, underline that caution must be exercised prior to analysis and interpretation of diet-induced effects on gene expression.
Highlights
The intestine is the site for nutrient digestion and absorption, and a major immunological defense barrier that recognizes and responds to external antigens
An overview of raw data for genes expressed in intestinal tissue from preterm pigs and in IPEC-J2 cells is presented in the heat map (Figure 1), based on color coding of the expression level before preprocessing
Including visual inspection of melting curves, 31 referenceand epithelial- and immune response-related genes were expressed within the dynamic range in pig intestinal tissue, and of these, 22 genes were expressed in IPEC-J2 cells
Summary
The intestine is the site for nutrient digestion and absorption, and a major immunological defense barrier that recognizes and responds to external antigens. It has previously been used to investigate hostpathogen interactions and immune responses with relevance for human and swine intestinal diseases [2, 3, 5,6,7,8] and could be a simple in vitro model to investigate the immune response of IECs in newborns. We set out to evaluate the IPEC-J2 cell line as an in vitro model for the preterm neonatal intestine and as a supporting model for the well-established preterm pig model of NEC, which is a valuable model for investigation of diet-induced effects [11,12,13,14]. In the preterm pig model of NEC, milk formula-fed preterm pigs have more NEC lesions compared with preterm pigs fed porcine or bovine colostrum, rich in growth- and immuno-modulatory factors [11, 15, 16]
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