Interleukin-23 Increases Intestinal Epithelial Cell Permeability In Vitro

Eur J Pediatr Surg. 2016 Jun;26(3):260-6. doi: 10.1055/s-0035-1551563. Epub 2015 May 22.

Abstract

Background Breast milk has a heterogeneous composition that differs between mothers and changes throughout the first weeks after birth. The proinflammatory cytokine IL-23 has a highly variable expression in human breast milk. We hypothesize that IL-23 found in human breast milk is biologically active and promotes epithelial barrier dysfunction. Methods The immature rat small intestinal epithelial cell line, IEC-18, was grown on cell inserts or standard cell culture plates. Confluent cultures were exposed to human breast milk with high or low levels of IL-23 and barrier function was measured using a flux of fluorescein isothiocyanate-dextran (FD-70). In addition, protein and mRNA expression of occludin and ZO-1 were measured and immunofluorescence used to stain occludin and ZO-1. Results Exposure to breast milk with high levels of IL-23 caused an increase flux of FD-70 compared with both controls and breast milk with low levels of IL-23. The protein expression of ZO-1 but not occludin was decreased by exposure to high levels of IL-23. These results correlate with immunofluorescent staining of ZO-1 and occludin which show decreased staining of occludin in both the groups exposed to breast milk with high and low IL-23. Conversely, cells exposed to high IL-23 breast milk had little peripheral staining of ZO-1 compared with controls and low IL-23 breast milk. Conclusion IL-23 in human breast milk is biologically active and negatively affects the barrier function of intestinal epithelial cells through the degradation of tight junction proteins.

MeSH terms

  • Animals
  • Cell Line
  • Cell Membrane Permeability / drug effects*
  • Dextrans / metabolism
  • Epithelial Cells / drug effects
  • Epithelial Cells / metabolism
  • Female
  • Fluorescein-5-isothiocyanate / analogs & derivatives
  • Fluorescein-5-isothiocyanate / metabolism
  • Humans
  • Interleukin-23 / analysis
  • Interleukin-23 / pharmacology*
  • Intestine, Small / cytology
  • Intestine, Small / physiology
  • Milk, Human / chemistry*
  • Occludin / genetics
  • Occludin / metabolism*
  • RNA, Messenger / metabolism
  • Rats
  • Real-Time Polymerase Chain Reaction
  • Zonula Occludens-1 Protein / metabolism*

Substances

  • Dextrans
  • Interleukin-23
  • Occludin
  • RNA, Messenger
  • Tjp1 protein, rat
  • Zonula Occludens-1 Protein
  • fluorescein isothiocyanate dextran
  • Fluorescein-5-isothiocyanate