INFLAMMATORY BOWEL DISEASE

Downregulation of epithelial apoptosis and barrier repair in active Crohn’s disease by tumour necrosis factor antibody treatment

S Zeissig^1,*, C Bojarski^1,*, N Buergel¹, J Mankertz¹, M Zeitz¹, M Fromm², J D Schulzke¹

¹ Departments of Gastroenterology, Infectious Diseases, and Rheumatology, Charité Campus Benjamin Franklin, Berlin, Germany
² Department of Clinical Physiology, Charité Campus Benjamin Franklin, Berlin, Germany

Correspondence to:
Correspondence to:
Professor J D Schulzke
Medizinische Klinik I, Charitè-Universitary Medicine Berlin, Campus Benjamin Franklin, Hindenburgdamm 30, Berlin 12200, Germany; joerg.schulzke{at}charite.de

Background and aims: Barrier dysfunction is an important feature contributing to inflammation and diarrhoea in Crohn’s disease (CD). Recently, tumour necrosis factor (TNF-) antibodies were recognised as effective in steroid refractory CD. The aim of this study was to characterise the effects of this therapy on the epithelial barrier.

Patients and methods: Forceps biopsies were obtained from the sigmoid colon before and 14 days after TNF- antibody therapy in 11 patients treated for chronic active CD (Crohn’s disease activity index >150). Epithelial apoptoses were measured after terminal deoxynucleotidyl transferase mediated deoxyuridine triphosphate nick end labelling (TUNEL) and 4',6-diamidino-2-phenylindole staining. Epithelial resistance was determined by alternating current impedance analysis in miniaturised Ussing chambers. Occludin, claudin 1, and claudin 4 expression was quantified in immunoblots.

Results: The epithelial apoptotic ratio was 2.1 (0.2)% in controls and increased to 5.3 (1.0)% in CD. TNF- antibody therapy decreased the apoptotic ratio to 2.9 (1.0)% (normalised in 10 of 11 patients). In parallel, epithelial resistance was lower in CD than in controls (24 (3) v 42 (3) xcm²) and improved to 34 (3) xcm² after therapy. Occludin, claudin 1, and claudin 4 were not affected by TNF- antibody therapy. In support of a functional role of epithelial apoptoses in CD, a similar decrease in resistance of –40% was observed when the apoptotic rate was selectively upregulated from 2.6% to 5.4% with camptothecin in HT-29/B6 cells.

Conclusions: Epithelial apoptoses were upregulated in the colon in CD and restored to normal in 10 of 11 patients by TNF- antibody therapy. This is the structural correlate of epithelial barrier dysfunction measured as epithelial resistance while expression of tight junction proteins did not contribute to this therapeutic effect.

Abbreviations: TNF-, tumour necrosis factor ; DAPI, 4',6-diamidino-2-phenylindole; R^e, epithelial electrical wall resistance; R^sub, subepithelial electrical wall resistance; R^t, transmural wall resistance; CD, Crohn’s disease; TUNEL, terminal deoxynucleotidyl transferase mediated deoxyuridine triphosphate nick end labelling; CDAI, Crohn’s disease activity index

Keywords: epithelial apoptosis; barrier repair; Crohn’s disease; tumour necrosis factor

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