title: Functional modulation of Crohn's disease myofibroblasts by anti-tumor necrosis factor antibodies.
abstract: OBJECTIVE
  Infliximab induces immune cell apoptosis by outside-to-inside signaling through transmembrane tumor necrosis factor-alpha (mTNF). However, in inflamed gut, myofibroblasts also produce TNF-alpha, and the affects of anti-TNF antibodies on these structural cells are unknown. We investigated the action of infliximab on apoptosis, the production of matrix metalloproteinases (MMPs) and tissue inhibitor of metalloproteinases (TIMP)-1, and migration of Crohn's disease (CD) myofibroblasts.
  
  
  METHODS
  Colonic myofibroblasts were isolated from patients with active CD and controls. mTNF was evaluated by Western blotting and flow cytometry. Infliximab-treated myofibroblasts were analyzed for apoptosis by Annexin V staining and caspase-3. TIMP-1 and MMPs were measured by Western blotting, and fibroblast migration was assessed by using an in vitro wound-healing scratch assay.
  
  
  RESULTS
  CD myofibroblasts showed higher mTNF expression than control myofibroblasts. Infliximab had no effect on CD myofibroblast apoptosis, caspase-3 activation, and production of MMP-3 and MMP-12. However, infliximab induced a significant dose-dependent increase in TIMP-1 production, which was inhibited by the p38 mitogen-activated protein kinase inhibitor SB 203580. The anti-TNF agents adalimumab, etanercept, and p55 TNF-receptor-human IgG fusion protein also increased TIMP-1 production. The migration of CD myofibroblasts was enhanced significantly by infliximab and recombinant human TIMP-1, and infliximab-induced migration was inhibited by anti-TIMP-1 neutralizing antibody. Infliximab also decreased CD myofibroblast collagen production.
  
  
  CONCLUSIONS
  Our findings show a novel therapeutic pathway for anti-TNF therapies in enhancing TIMP-1 production and myofibroblast migration, which may reduce MMP activity and facilitate the wound healing.
authors.affiliation: Centre for Infectious Disease, Institute of Cell and Molecular Science, Barts and the London Queen Mary's School of Medicine and Dentistry, London, United Kingdom.
authors.forename: Antonio
authors.initials: A
authors.lastname: Di Sabatino
authors.forename: Sylvia L F
authors.initials: SL
authors.lastname: Pender
authors.forename: Claire L
authors.initials: CL
authors.lastname: Jackson
authors.forename: Joanna D
authors.initials: JD
authors.lastname: Prothero
authors.forename: John N
authors.initials: JN
authors.lastname: Gordon
authors.forename: Lucia
authors.initials: L
authors.lastname: Picariello
authors.forename: Laura
authors.initials: L
authors.lastname: Rovedatti
authors.forename: Guillermo
authors.initials: G
authors.lastname: Docena
authors.forename: Giovanni
authors.initials: G
authors.lastname: Monteleone
authors.forename: David S
authors.initials: DS
authors.lastname: Rampton
authors.forename: Francesco
authors.initials: F
authors.lastname: Tonelli
authors.forename: Gino R
authors.initials: GR
authors.lastname: Corazza
authors.forename: Thomas T
authors.initials: TT
authors.lastname: MacDonald
mesh_terms: D000328:Adult; D000893:Anti-Inflammatory Agents / Q000494:pharmacology; D000911:Antibodies, Monoclonal / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects / Q000276:immunology; D001323:Autoantibodies / Q000276:immunology / Q000494:pharmacology*; D001706:Biopsy; D002478:Cells, Cultured; D003094:Collagen / Q000378:metabolism; D003106:Colon / Q000276:immunology / Q000473:pathology; D003424:Crohn Disease / Q000276:immunology* / Q000473:pathology*; D005347:Fibroblasts / Q000276:immunology* / Q000378:metabolism / Q000473:pathology; D006801:Humans; D000069285:Infliximab; D053508:Matrix Metalloproteinase 12 / Q000378:metabolism; D019278:Matrix Metalloproteinase 3 / Q000378:metabolism; D008875:Middle Aged; D009924:Organ Culture Techniques; D019715:Tissue Inhibitor of Metalloproteinase-1 / Q000276:immunology / Q000378:metabolism / Q000494:pharmacology; D014409:Tumor Necrosis Factor-alpha / Q000276:immunology*; D014945:Wound Healing / Q000187:drug effects / Q000276:immunology
chemical_list: D000893:Anti-Inflammatory Agents; D000911:Antibodies, Monoclonal; D001323:Autoantibodies; D019715:Tissue Inhibitor of Metalloproteinase-1; D014409:Tumor Necrosis Factor-alpha; D003094:Collagen; D000069285:Infliximab; D019278:Matrix Metalloproteinase 3; D053508:Matrix Metalloproteinase 12