Functional modulation of Crohn's disease myofibroblasts by anti-tumor necrosis factor antibodies.
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.
Di Sabatino
Antonio
A
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.
Pender
Sylvia L F
SL
Jackson
Claire L
CL
Prothero
Joanna D
JD
Gordon
John N
JN
Picariello
Lucia
L
Rovedatti
Laura
L
Docena
Guillermo
G
Monteleone
Giovanni
G
Rampton
David S
DS
Tonelli
Francesco
F
Corazza
Gino R
GR
MacDonald
Thomas T
TT
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
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