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Updated: Jul 18, 2026
A Novel Human Epithelial Enteroid Model of Necrotizing Enterocolitis
Published on: April 10, 2019
In vitro model of intestinal crypt abscess. A novel neutrophil-derived secretagogue activity
1Department of Pathology, Brigham and Women's Hospital, Boston, Massachusetts.
Abstract:
In order to model crypt abscesses, a histological finding which correlates with disease activity in intestinal inflammation, human polymorphonuclear leukocytes (PMN) were layered onto monolayers of the human intestinal epithelial cell line T84, a crypt-like epithelium which is capable of Cl- secretion. Such PMN-epithelial interaction had no substantial effect on monolayer integrity or function. However, when PMN were stimulated by conditions including those present naturally in the human colonic lumen, monolayers responded with a bumetanide-sensitive short circuit current (Isc) indicative of Cl- secretion, the basis of secretory diarrhea. This Isc response was induced by a neutrophil-derived secretagogue (NDS), which was only active when applied to the luminal surface of monolayers and did not require PMN-epithelial contact. NDS activity is resistant to boiling, acid, and trypsin and passes a 500 nominal mol wt cutoff filter. NDS activity is not secondary to the respiratory burst products O2- or H2O2 and does not appear to be a myeloperoxidase product. We speculate NDS elicited Cl- secretion may contribute to the secretory diarrhea seen in patients with intestinal inflammation and crypt abscesses.
Insights
Human neutrophils release a factor that causes intestinal cells to secrete chloride, potentially explaining secretory diarrhea in inflammatory bowel disease with crypt abscesses.
Area of Science:
- Gastroenterology
- Immunology
- Cell Biology
Background:
- Crypt abscesses are histological markers of intestinal inflammation.
- Intestinal inflammation can lead to secretory diarrhea.
- Human intestinal epithelial cells (T84) form crypt-like monolayers.
Purpose of the Study:
- To model crypt abscesses and their role in intestinal inflammation.
- To investigate the interaction between human polymorphonuclear leukocytes (PMN) and intestinal epithelial cells.
- To identify mechanisms of chloride secretion in response to PMN stimulation.
Main Methods:
- Layering human PMN onto T84 cell monolayers.
- Stimulating PMN under conditions mimicking the colonic lumen.
- Measuring short-circuit current (Isc) to assess chloride secretion.
- Characterizing the properties of the neutrophil-derived secretagogue (NDS).
Main Results:
- PMN-epithelial interactions did not affect monolayer integrity.
- Stimulated PMN induced bumetanide-sensitive Isc, indicating chloride secretion.
- A neutrophil-derived secretagogue (NDS) was identified, acting on the luminal surface.
- NDS activity is heat, acid, and trypsin resistant, and filterable (500 Da cutoff).
- NDS is not derived from respiratory burst products or myeloperoxidase.
Conclusions:
- A neutrophil-derived secretagogue may contribute to secretory diarrhea in intestinal inflammation.
- NDS-induced chloride secretion is a potential mechanism in conditions like crypt abscesses.
- Further research is needed to elucidate the exact nature and role of NDS.
