title: Soluble galectin-3 is a strong, colonic epithelial-cell-derived, lamina propria fibroblast-stimulating factor.
abstract: BACKGROUND
  Colonic lamina propria fibroblasts (CLPFs) play an important role in the pathogenesis of fibrosis and strictures in Crohn's disease.
  
  
  OBJECTIVE
  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 kappaB (NF-kappaB) was shown, and translocation of NF-kappaB was inhibited by a dominant-negative IkappaB-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/microg v 1.434 (0.695) pg/microg) was found. The effect of CEC-conditioned media on CLPF IL8 secretion was NF-kappaB dependent. A protein or DNA array confirmed the involvement of NF-kappaB 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-kappaB activation and IL8 secretion in these cells and may be a target for future therapeutic approaches to reduce or avoid stricture formation.
authors.affiliation: Department of Internal Medicine I, University of Regensburg, Regensburg 93042, Germany.
authors.forename: E
authors.initials: E
authors.lastname: Lippert
authors.forename: W
authors.initials: W
authors.lastname: Falk
authors.forename: F
authors.initials: F
authors.lastname: Bataille
authors.forename: T
authors.initials: T
authors.lastname: Kaehne
authors.forename: M
authors.initials: M
authors.lastname: Naumann
authors.forename: M
authors.initials: M
authors.lastname: Goeke
authors.forename: H
authors.initials: H
authors.lastname: Herfarth
authors.forename: J
authors.initials: J
authors.lastname: Schoelmerich
authors.forename: G
authors.initials: G
authors.lastname: Rogler
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D018938:Caco-2 Cells; D002460:Cell Line; D002478:Cells, Cultured; D003106:Colon / Q000737:chemistry*; D015179:Colorectal Neoplasms / Q000473:pathology; D017077:Culture Media, Conditioned; D004238:Diverticulitis / Q000473:pathology; D004847:Epithelial Cells / Q000737:chemistry; D005260:Female; D005347:Fibroblasts / Q000187:drug effects*; D037502:Galectin 3 / Q000032:analysis*; D019073:HT29 Cells; D006801:Humans; D016209:Interleukin-8 / Q000032:analysis; D007410:Intestinal Diseases / Q000473:pathology; D008297:Male; D008875:Middle Aged; D009092:Mucous Membrane / Q000737:chemistry*; D016328:NF-kappa B / Q000378:metabolism; D018808:Transcription Factor AP-1 / Q000378:metabolism
chemical_list: D017077:Culture Media, Conditioned; D037502:Galectin 3; D016209:Interleukin-8; D016328:NF-kappa B; D018808:Transcription Factor AP-1