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Activation of NF-κB in intestinal epithelial cells by E. coli strains isolated from the colonic mucosa of IBD patients

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Abstract

Background and aims

The involvement of bacteria in the pathogenesis of inflammatory bowel disease has been discussed for several years. In this study we evaluated the ability of E. coli isolates from inflamed and noninflamed colonic mucosa to activate NF-κB.

Materials and methods

Fifteen bacterial strains from inflamed and six from noninflamed colonic tissues from IBD patients. Their ability to induce NF-κB activation was examined in vitro by gel-shift assays. The activation of the TNF-α promoter was determined by reporter gene assays. Bacterial isolates were characterized by invasion assays, electron microscopy, and PCR.

Results

Four of 15 E. coli bacterial isolates from inflamed IBD tissues induced NF-κB activity in intestinal epithelial cells as determined by gel-shift assays. NF-κB activation was only seen with living bacteria but not with heat-inactivated cells. Isolates from noninflamed tissues and a wild-type E. coli control strain induced a weaker or no activation. Reporter gene assays with a construct comprising a luciferase gene driven by the TNF-α promoter revealed that isolates from Crohn’s disease patients induced a stronger activation of the TNF-α gene than isolates from ulcerative colitis patients. The isolated bacteria invaded HT-29 cells, although typical virulence genes for enteropathogenic, enterhemorrhagic, or enteroinvasive E. coli, i.e., eae, tir, EspA, Per (A-C), ipaC, were not detected in these cells. Bacterial invasion was additionally confirmed by electron microscopy examination.

Conclusion

Our results indicate that E. coli strains can be found in the mucosa of some IBD patients which are able to activate NF-κB similar to known pathogenic strains. The absence of several virulence genes in these cells suggests that they are members of the luminal flora which acquire as yet unidentified virulence determinants and are therefore involved in the pathophysiology of IBD.

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Acknowledgements

This work was supported in part by grants from a TMR project and by SFB415. We acknowledge the help of the nurses in the endoscopy department for their excellent work as well as Dr. Stephan Ott, Meike Barche, and Lorena Vailles for their skillful technical assistance. We also express many thanks to Marie-Luise Kruse who supported the preparation of the microscope samples.

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Authors and Affiliations

  1. Department of Pediatrics, University of Ulm, Prittwitzstrasse 43, 89075, Ulm, Germany

    Katia La Ferla

  2. CONARIS Research Institute AG, Schauenburgerstrasse 116, 24118, Kiel, Germany

    Dirk Seegert

  3. 1st Department of Medicine, University Clinic of Schleswig-Holstein, Schittenhelmstrasse 12, 24105, Kiel, Germany

    Dirk Seegert & Stefan Schreiber

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  1. Katia La Ferla
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  2. Dirk Seegert
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Correspondence to Dirk Seegert.

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La Ferla, K., Seegert, D. & Schreiber, S. Activation of NF-κB in intestinal epithelial cells by E. coli strains isolated from the colonic mucosa of IBD patients. Int J Colorectal Dis 19, 334–342 (2004). https://doi.org/10.1007/s00384-004-0583-7

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