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Eosinophil granulocytes are activated during the remission phase of ulcerative colitis
M Lampinen1, A Rönnblom, K Amin
1Department of Medical Sciences, Gastroenterology Group, Clinical Chemistry and Medicine, University Hospital, S-751 85 Uppsala, Sweden. maria.lampinen@medsci.uu.se
Insights
Activated eosinophils are found in both active and inactive ulcerative colitis (UC), suggesting a role in inflammation and potentially epithelial repair. Neutrophils are active during active UC but not in remission.
Area of Science:
- Gastroenterology
- Immunology
- Cell Biology
Background:
- Ulcerative colitis (UC) involves chronic inflammation of the gastrointestinal tract.
- The roles of eosinophils and neutrophils in UC pathogenesis are not fully elucidated.
- Understanding immune cell activity in UC is crucial for developing targeted therapies.
Purpose of the Study:
- To develop a flow cytometry method for analyzing intestinal eosinophils and neutrophils.
- To compare the distribution and activation of these cells in different stages and locations of UC.
- To investigate the cellular mechanisms underlying UC and its remission.
Main Methods:
- Biopsy samples were collected from multiple colonic locations and the terminal ileum in UC patients and controls.
- Cell suspensions were prepared and incubated with fluorescently labeled monoclonal antibodies.
- Flow cytometry with scatter plot-gating and specific antibodies was employed to quantify and assess cell activity.
Main Results:
- Eosinophils were more numerous and active in active UC compared to controls, with even higher activation during inactive UC.
- Neutrophils increased in number and activity during active UC but not inactive UC.
- Activated eosinophils were prominent in the rectum in distal colitis, while activated neutrophils were localized to the sigmoid colon and rectum.
Conclusions:
- The developed flow cytometry method effectively characterizes intestinal granulocytes in UC.
- Neutrophils are key players in active UC inflammation, exhibiting a resting state during remission.
- Activated eosinophils in UC suggest both pro-inflammatory and potential epithelial repair roles, particularly during inactive disease.
Aim:
The aim of this study was to establish a method of investigating intestinal eosinophil and neutrophil granulocytes by flow cytometry, and to compare the distribution and activity of these cells in different stages of ulcerative colitis (UC).
Methods:
Biopsy samples were taken from six locations of the entire colon and from the terminal ileum in 10 patients with active total UC, 10 patients with inactive total UC, eight patients with active distal UC, and 11 control subjects. Cell suspensions from biopsies and from peripheral blood were incubated with fluorophore conjugated monoclonal antibodies. The use of scatter plot-gating and specific antibodies was established in a flow cytometry assay.
Results:
Eosinophils were more numerous and more active in patients with active UC than in controls. Interestingly, during inactive UC, the number of activated eosinophils was even larger. Eosinophil activity was high in the rectum of patients with distal colitis but was also slightly elevated in the proximal colon. Neutrophils were increased in number and activity during active but not inactive UC. In patients with distal colitis, activated neutrophils were only found in the sigmoid colon and rectum.
Conclusion:
With this method, we confirm that neutrophils participate in the inflammatory process during active UC, and that they express a resting phenotype during remission. The finding of activated eosinophils in inflamed intestine strengthens the view of these cells as proinflammatory and tissue damaging. Nevertheless, our new finding of high eosinophil activation during inactive UC suggests that eosinophils play a role in repair of injured epithelium.
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