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Deoxynivalenol affects the composition of the basement membrane proteins and influences en route the migration of CD16(+) cells into the intestinal epithelium.

Nossol C, Diesing AK, Kahlert S, Kersten S, Kluess J, Ponsuksili S, Hartig R, Wimmers K, Dänicke S, Rothkötter HJ - Mycotoxin Res (2013)

Bottom Line: The in vivo results were extended with microarray analyses of epithelial cell (IPEC-J2 cells).Our results provide evidence that already low basolateral concentrations of DON (50 ng/mL) influence the production of the BM protein laminin by epithelial cells.Thus, DON affects the composition of the BM.

View Article: PubMed Central - PubMed

Affiliation: Institute of Anatomy, Otto-von-Guericke University Magdeburg, Leipziger Strasse 44, 39120, Magdeburg, Germany, constanze.nossol@med.ovgu.de.

ABSTRACT
The numerous pores in the basement membrane (BM) of the intestinal villi are essential for the communication of enterocytes with cells in the lamina propria, an important mechanism for the induction of intestinal immune responses. The intestinal epithelial barrier is affected by the mycotoxin deoxynivalenol (DON) from both the apical (luminal) and basolateral (serosal) side. The pig is the most susceptible species to the anorectic and immune-modulating effects of DON, which is most prevalent in crops. We analysed in pigs the effect of DON-contaminated feed on the composition and perforation of the BM and the presence of CD16(+) cells or their dendrites in the epithelium. In addition to in vivo experiments, in vitro studies were carried out. Using microarray analyses, the effects of DON on IPEC-J2 cells were studied with the focus on the BM. Our in vivo results showed in the control pigs: (1) a significant increased pore number (p ≤ 0.001) in the jejunum in comparison to ileum, (2) no difference in the pore size, and (3) comparable frequency of intraepithelial CD16(+) cells/dendrites in the jejunum and ileum. There was a marked trend that DON feeding increases: (1) the pore number in jejunum, and (2) the number of CD16(+) cells/dendrites in the epithelium (Tukey-Kramer; p = 0.055 and p = 0.067, respectively). The in vivo results were extended with microarray analyses of epithelial cell (IPEC-J2 cells). The down-regulation of genes like syndecan, fibulin 6 and BM-40 was observed. These proteins are important factors in the BM composition and in formation of pores. Our results provide evidence that already low basolateral concentrations of DON (50 ng/mL) influence the production of the BM protein laminin by epithelial cells. Thus, DON affects the composition of the BM.

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Analysis of the laminin expression on mRNA-level using to qPCR. IPEC-J2 cells were treated with DON 200 ng/mL ap and 2,000 ng/mL ap which resemble the chronic and the acute DON-exposure. The low concentration of 50 ng/mL was used to reflect the DON-concentration found in the peripheral blood after feeding DON-diet in pigs (Dänicke et al. 2004). No differences were found in the comparison of control and DON-treatment groups (RQ relative quantification)
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Fig7: Analysis of the laminin expression on mRNA-level using to qPCR. IPEC-J2 cells were treated with DON 200 ng/mL ap and 2,000 ng/mL ap which resemble the chronic and the acute DON-exposure. The low concentration of 50 ng/mL was used to reflect the DON-concentration found in the peripheral blood after feeding DON-diet in pigs (Dänicke et al. 2004). No differences were found in the comparison of control and DON-treatment groups (RQ relative quantification)

Mentions: As the effects of DON 200 ng/mL induced diverse effects in the laminin precursors expression, additional qPCR analyses were carried out in IPEC-J2 and the laminin α-3 chain precursor (LAMA3) and laminin γ-1 chain precursor (LAMC1) were studied. To check the effect of an even lower DON concentration (comparable to the level that has been observed in the peripheral blood), DON 50 ng/mL bl was used instead of DON 200 ng/mL bl. No significant differences in the LAMA3 and the LAMC1 mRNA-level were found in the DON-treatment groups in comparison to control (Fig. 7).Fig. 7


Deoxynivalenol affects the composition of the basement membrane proteins and influences en route the migration of CD16(+) cells into the intestinal epithelium.

Nossol C, Diesing AK, Kahlert S, Kersten S, Kluess J, Ponsuksili S, Hartig R, Wimmers K, Dänicke S, Rothkötter HJ - Mycotoxin Res (2013)

Analysis of the laminin expression on mRNA-level using to qPCR. IPEC-J2 cells were treated with DON 200 ng/mL ap and 2,000 ng/mL ap which resemble the chronic and the acute DON-exposure. The low concentration of 50 ng/mL was used to reflect the DON-concentration found in the peripheral blood after feeding DON-diet in pigs (Dänicke et al. 2004). No differences were found in the comparison of control and DON-treatment groups (RQ relative quantification)
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3824239&req=5

Fig7: Analysis of the laminin expression on mRNA-level using to qPCR. IPEC-J2 cells were treated with DON 200 ng/mL ap and 2,000 ng/mL ap which resemble the chronic and the acute DON-exposure. The low concentration of 50 ng/mL was used to reflect the DON-concentration found in the peripheral blood after feeding DON-diet in pigs (Dänicke et al. 2004). No differences were found in the comparison of control and DON-treatment groups (RQ relative quantification)
Mentions: As the effects of DON 200 ng/mL induced diverse effects in the laminin precursors expression, additional qPCR analyses were carried out in IPEC-J2 and the laminin α-3 chain precursor (LAMA3) and laminin γ-1 chain precursor (LAMC1) were studied. To check the effect of an even lower DON concentration (comparable to the level that has been observed in the peripheral blood), DON 50 ng/mL bl was used instead of DON 200 ng/mL bl. No significant differences in the LAMA3 and the LAMC1 mRNA-level were found in the DON-treatment groups in comparison to control (Fig. 7).Fig. 7

Bottom Line: The in vivo results were extended with microarray analyses of epithelial cell (IPEC-J2 cells).Our results provide evidence that already low basolateral concentrations of DON (50 ng/mL) influence the production of the BM protein laminin by epithelial cells.Thus, DON affects the composition of the BM.

View Article: PubMed Central - PubMed

Affiliation: Institute of Anatomy, Otto-von-Guericke University Magdeburg, Leipziger Strasse 44, 39120, Magdeburg, Germany, constanze.nossol@med.ovgu.de.

ABSTRACT
The numerous pores in the basement membrane (BM) of the intestinal villi are essential for the communication of enterocytes with cells in the lamina propria, an important mechanism for the induction of intestinal immune responses. The intestinal epithelial barrier is affected by the mycotoxin deoxynivalenol (DON) from both the apical (luminal) and basolateral (serosal) side. The pig is the most susceptible species to the anorectic and immune-modulating effects of DON, which is most prevalent in crops. We analysed in pigs the effect of DON-contaminated feed on the composition and perforation of the BM and the presence of CD16(+) cells or their dendrites in the epithelium. In addition to in vivo experiments, in vitro studies were carried out. Using microarray analyses, the effects of DON on IPEC-J2 cells were studied with the focus on the BM. Our in vivo results showed in the control pigs: (1) a significant increased pore number (p ≤ 0.001) in the jejunum in comparison to ileum, (2) no difference in the pore size, and (3) comparable frequency of intraepithelial CD16(+) cells/dendrites in the jejunum and ileum. There was a marked trend that DON feeding increases: (1) the pore number in jejunum, and (2) the number of CD16(+) cells/dendrites in the epithelium (Tukey-Kramer; p = 0.055 and p = 0.067, respectively). The in vivo results were extended with microarray analyses of epithelial cell (IPEC-J2 cells). The down-regulation of genes like syndecan, fibulin 6 and BM-40 was observed. These proteins are important factors in the BM composition and in formation of pores. Our results provide evidence that already low basolateral concentrations of DON (50 ng/mL) influence the production of the BM protein laminin by epithelial cells. Thus, DON affects the composition of the BM.

Show MeSH
Related in: MedlinePlus