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Related Experiment Video

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Investigating Intestinal Inflammation in DSS-induced Model of IBD
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Lysozyme gene expression in inflammatory bowel disease.

G W Stamp1, R Poulsom, L P Chung

  • 1Histopathology Unit in Sity Hybridisation Laboratory, Imperial Cancer Research Fund, Lincoln's Inn Fields, London.

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Riboprobe in situ hybridization reveals active lysozyme synthesis in inflammatory bowel diseases. Neutrophils contribute to fecal lysozyme in ulcerative colitis, while macrophages elevate serum lysozyme in Crohn's disease.

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Area of Science:

  • Gastroenterology
  • Molecular Biology
  • Immunology

Background:

  • Inflammatory bowel diseases (IBD), including ulcerative colitis (UC) and Crohn's disease (CD), involve complex immune responses.
  • Lysozyme, an antimicrobial enzyme, plays a role in innate immunity and is implicated in IBD pathogenesis.
  • Previous studies using immunohistochemistry have provided insights but can be limited by cell infiltration and enzyme localization.

Purpose of the Study:

  • To investigate the sites of active lysozyme synthesis in ulcerative colitis and Crohn's disease using riboprobe in situ hybridization (rISH).
  • To differentiate lysozyme expression patterns between UC and CD and identify specific cell types involved in its production.
  • To clarify the cellular sources of elevated lysozyme levels observed in fecal and serum samples from IBD patients.

Main Methods:

  • Riboprobe in situ hybridization (rISH) was employed to detect lysozyme messenger RNA (mRNA) in tissue samples from patients with ulcerative colitis and Crohn's disease.
  • Immunohistochemical studies were used for comparison, highlighting the limitations of detecting cellular infiltration.
  • Analysis focused on identifying maximal labeling in specific cell populations such as Paneth cells, macrophages, and granulomas.

Main Results:

  • rISH demonstrated active lysozyme synthesis in both ulcerative colitis and Crohn's disease, with maximal labeling in Paneth cells, macrophages, and granulomas.
  • In ulcerative colitis, strong lysozyme mRNA signals were found in Paneth cell metaplasia and regenerative mucosa, while neutrophils did not express lysozyme mRNA.
  • In Crohn's disease, abundant lysozyme mRNA was observed in tuberculoid granulomas and macrophage aggregates, distinct patterns not seen in ulcerative colitis.

Conclusions:

  • rISH provides a clearer picture of active lysozyme synthesis in the gut during IBD compared to immunohistochemistry, especially in the presence of polymorph infiltration.
  • Neutrophils are likely the primary source of elevated fecal lysozyme in ulcerative colitis.
  • Macrophages are the probable source of elevated serum lysozyme in Crohn's disease.