Alimentary TractThe intestinal mucus layer from patients with inflammatory bowel disease harbors high numbers of bacteria compared with controls☆,☆☆
Section snippets
Patients and controls
Nineteen consecutive patients with IBD visiting the Department of Gastroenterology for endoscopy were included. Patients were enrolled in the study if they had clinically active UC, CD, or indeterminate colitis. Diagnosis was confirmed by endoscopy and histology. Controls were 9 patients who required endoscopy for reasons other than IBD (e.g., irritable bowel syndrome) and who did not complain of diarrhea. Controls were only included if results from endoscopy and histological analysis of biopsy
ISH of sheep collagen-colon cell specimens
Sections of sheep collagen specimens containing HT-29 cells and different species of Enterobacteriaceae, H. pylori, and S. aureus were hybridized with probes Eub338M, dT35f, and EB1f. After hybridization with Eub338M, bacteria of all species tested stained bright brown in contrast to the HT29-19A cells and the collagen in the tissue (Figure 1A).
Discussion
In this study we investigated the spatial distribution of intestinal bacteria in the rectal and colonic mucosa in paraffin-embedded biopsy specimens from 19 patients with IBD and from 14 controls. We developed a nonradioactive method for ISH enabling us to study the distribution of intestinal bacteria by light microscopy. To detect as many bacteria as possible, we used a probe that has been shown to detect rRNA of eubacteria present in the human large intestine.12, 13 In our ISH reconstruction
Acknowledgements
The authors thank P. van Amstel en W. van Est for technical assistance, P. Fockens for assistance with sample collection, R. van Leeuwen for assistance with statistical analyses, and M. D. de Jong for critically reviewing the manuscript.
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Address requests for reprints to: Constance Schultsz, M.D., Department of Medical Microbiology, Academic Medical Center, Meibergdreef 9, 1105 AZ Amsterdam, The Netherlands. e-mail: [email protected].
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Dr. Cohen has worked as a consultant to Centocor, Inc. Dr. Cominelli has a grant from Centocor, Inc. to study the incidence of lymphoma among hospitalized Crohn's disease patients. He is also a member of the speaking bureau for Centocor, Inc. Drs. Cominelli and Bickston are currently clinical trial investigators of CDP571 (CellTech Therapeutic Ltd., Berkshire, England).