Functional Modulation of Crohn’s Disease Myofibroblasts by Anti-Tumor Necrosis Factor Antibodies

ANTONIO DI SABATINO; SYLVIA L. F. PENDER; CLAIRE L. JACKSON; JOANNA D. PROTHERO; JOHN N. GORDON; LUCIA PICARIELLO; LAURA ROVEDATTI; GUILLERMO DOCENA; GIOVANNI MONTELEONE; DAVID S. RAMPTON; FRANCESCO TONELLI; GINO R. CORAZZA; THOMAS T. MACDONALD

GASTROENTEROLOGY

W B SAUNDERS CO-ELSEVIER INC

Año: 2007 vol. 133 p. 137 - 149

0016-5085

Background & Aims: Infliximab induces immune cellapoptosis by outside-to-inside signaling through transmembranetumor necrosis factor- (mTNF). However,in inflamed gut, myofibroblasts also produce TNF-,and the affects of anti-TNF antibodies on these structuralcells are unknown. We investigated the action ofinfliximab on apoptosis, the production of matrixmetalloproteinases (MMPs) and tissue inhibitor of metalloproteinases(TIMP)-1, and migration of Crohn’sdisease (CD) myofibroblasts. Methods: Colonic myofibroblastswere isolated from patients with active CDand controls. mTNF was evaluated by Western blottingand flow cytometry. Infliximab-treated myofibroblastswere analyzed for apoptosis by Annexin V staining andcaspase-3. TIMP-1 and MMPs were measured by Westernblotting, and fibroblast migration was assessed byusing an in vitro wound-healing scratch assay. Results:CD myofibroblasts showed higher mTNF expressionthan control myofibroblasts. Infliximab had no effecton CD myofibroblast apoptosis, caspase-3 activation,and production of MMP-3 and MMP-12. However, infliximabinduced a significant dose-dependent increasein TIMP-1 production, which was inhibited by the p38mitogen-activated protein kinase inhibitor SB 203580.The anti-TNF agents adalimumab, etanercept, and p55TNF-receptor–human IgG fusion protein also increasedTIMP-1 production. The migration of CD myofibroblastswas enhanced significantly by infliximaband recombinant human TIMP-1, and infliximab-inducedmigration was inhibited by anti–TIMP-1 neutralizingantibody. Infliximab also decreased CD myofibroblastcollagen production. Conclusions: Our findingsshow a novel therapeutic pathway for anti-TNF therapiesin enhancing TIMP-1 production and myofibroblastmigration, which may reduce MMP activity andfacilitate the wound healing.