title: Intestinal trefoil factor induces decay-accelerating factor expression and enhances the protective activities against complement activation in intestinal epithelial cells.
abstract: Mucosal damage induces a massive influx of serum complement components into the lumen. The epithelium produces a number of factors that can potentially ameliorate injury including intestinal trefoil factor (ITF), a small protease-resistant peptide produced and secreted onto the mucosal surface by goblet cells, and decay-accelerating factor (DAF), a protein produced by columnar epithelium which protects the host tissue from autologous complement injury. However, coordination of these intrinsic defensive products has not been delineated. DAF protein and mRNA expression were evaluated by immunoblotting and Northern blotting, respectively. NF-kappaB-DNA binding activity and DAF promoter activity were assessed by an electrophoretic gel mobility shift assay and a reporter gene luciferase assay, respectively. ITF induced a dose- and time-dependent increase in DAF protein and mRNA expression in human (HT-29 and T84) and rat (IEC-6) intestinal epithelial cells. In differentiated T84 cells grown on cell culture inserts, basolateral stimulation with ITF strongly enhanced DAF expression, but apical stimulation had no effects. The C3 deposition induced by complement activation was significantly blocked by the treatment with ITF. In HT-29 cells, ITF increased the stability of DAF mRNA. ITF also enhanced the promoter activity of the DAF gene via NF-kappaB motif and induced activation of NF-kappaB-DNA binding activity. ITF promotes protection of epithelial cells from complement activation via up-regulation of DAF expression, contributing to a robust mucosal defense.
authors.affiliation: Gastrointestinal Unit, Department of Medicine, Massachusetts General Hospital, 55 Fruit Street, Boston, MA 02114, USA.
authors.forename: A
authors.initials: A
authors.lastname: Andoh
authors.forename: K
authors.initials: K
authors.lastname: Kinoshita
authors.forename: I
authors.initials: I
authors.lastname: Rosenberg
authors.forename: D K
authors.initials: DK
authors.lastname: Podolsky
mesh_terms: D000818:Animals; D018958:CD55 Antigens / Q000096:biosynthesis / Q000235:genetics*; D002460:Cell Line; D003167:Complement Activation; D003176:Complement C3 / Q000378:metabolism; D006133:Growth Substances / Q000494:pharmacology*; D006801:Humans; D018928:Immunity, Mucosal; D007413:Intestinal Mucosa / Q000276:immunology*; D007700:Kinetics; D009077:Mucins; D009124:Muscle Proteins; D016328:NF-kappa B / Q000378:metabolism; D009479:Neuropeptides; D010455:Peptides / Q000494:pharmacology*; D011401:Promoter Regions, Genetic; D014176:Protein Biosynthesis; D020871:RNA Stability; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D015533:Transcriptional Activation; D000071164:Trefoil Factor-2; D000071165:Trefoil Factor-3; D014407:Tumor Cells, Cultured
chemical_list: D018958:CD55 Antigens; D003176:Complement C3; D006133:Growth Substances; D009077:Mucins; D009124:Muscle Proteins; D016328:NF-kappa B; D009479:Neuropeptides; D010455:Peptides; D012333:RNA, Messenger; C099037:TFF3 protein, rat; D000071164:Trefoil Factor-2; D000071165:Trefoil Factor-3