INFLAMMATORY BOWEL DISEASE

Soluble galectin-3 is a strong, colonic epithelial-cell-derived, lamina propria fibroblast-stimulating factor

E Lippert¹, W Falk¹, F Bataille², T Kaehne³, M Naumann³, M Goeke⁴, H Herfarth¹, J Schoelmerich¹, G Rogler¹

¹ Department of Internal Medicine I, University of Regensburg, Regensburg, Germany
² Insitute of Pathology, University of Regensburg, Regensburg, Germany
³ Institute of Experimental Internal Medicine, Otto-von-Guericke University, Magdeburg, Germany
⁴ Department of Gastroenterology, Hepatology and Endocrinology, Medizinische Hochschule, Hannover, Germany

Correspondence to:
Dr G Rogler
Department of Internal Medicine I, University of Regensburg, Regensburg 93042, Germany; gerhard.rogler{at}klinik.uni-regensburg.de

Background: Colonic lamina propria fibroblasts (CLPFs) play an important role in the pathogenesis of fibrosis and strictures in Crohn’s disease.

Aim: To identify colonic epithelial cell (CEC)-derived factors that activate CLPFs.

Methods: Primary human CECs and CLPFs were isolated from control mucosa and interleukin 8 (IL8) of CLPF cultures was quantified by ELISA. Activation of nuclear factor B (NF-B) was shown, and translocation of NF-B was inhibited by a dominant-negative IB-expressing adenovirus. The major CLPF-activating and IL8 inducing protein was purified using fast-performance liquid chromatography (HiPrep 16/60 Sephacryl S-200 High Resolution Column) and sodium dodecyl sulphate gel electrophoresis.

Results: A considerable increase in IL8 secretion by CLPFs cultured in CEC-conditioned media compared with that in unconditioned media (155.00 (10.00) pg/µg v 1.434 (0.695) pg/µg) was found. The effect of CEC-conditioned media on CLPF IL8 secretion was NF-B dependent. A protein or DNA array confirmed the involvement of NF-B and activator protein-1. Purification of a candidate band isolated with the use of sodium dodecyl sulphate-polyacrylamide gel electrophoresis and subsequent sequencing showed soluble galectin-3 to be a strong CLPF-activating factor. Depletion of galectin-3 from conditioned media by immunoprecipitation abolished the CLPF stimulatory effect.

Conclusions: Using a classical biochemical approach, soluble galectin-3 was identified as a strong activator of CLPFs produced by CEC. Galectin-3 induced NF-B activation and IL8 secretion in these cells and may be a target for future therapeutic approaches to reduce or avoid stricture formation.

Abbreviations: CEC, colonic epithelial cell; CLPF, colonic lamina propria fibroblast; CRD, carbohydrate recognition domain; DMEM, Dulbecco’s modified Eagle’s medium; FCS, fetal calf serum; FPLC, fast-performance liquid chromatography; IEC, intestinal epithelial cell; IB, inhibitor protein B; MALDI-TOF, matrix-assisted laser desorption/ionization time-of-flight; MOI, multiplicity of infection; NF-B, nuclear factor B; PBS, phosphate buffered saline; SDS-PAGE, sodium dodecyl sulphate-polyacrylamide gel electrophoresis; TGF, transforming growth factor

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