Enhanced human beta-defensin-2 (hBD-2) expression by corticosteroids is independent of NF-kappaB in colonic epithelial cells (CaCo2).
Beta-defensins are small cationic peptides with antimicrobial properties that contribute to innate host defense. Unlike human beta-defensin-1 (hBD-1), which is produced constitutively, human beta-defensin-2 (hBD-2) is expressed after adequate stimulation by cytokines and/or bacterial endotoxins in epithelial tissue and mononuclear phagocytes but may be deficient in patients with Crohn's disease. To further elucidate the role of the intestinal epithelium in antimicrobial host defense, gene regulation of hBD-2 and the interaction with NF-kappaB were analyzed in a cell culture model. Human colonic epithelial cells (CaCo2) were stimulated by pro-inflammatory cytokines (IL-1beta, TNF-alpha, IF-gamma) to induce hBD-2 mRNA transcription. Interactions with NF-kappaB were analyzed using specific inhibitors (sulfasalazine, gliotoxine, dexamethasone) at different concentrations. Defensin mRNA expression was quantified by competitive RT-PCR and antibacterial capacity of supernatants was determined by an antimicrobial assay. HBD-2 mRNA transcription and antimicrobial activity of CaCo2 cells were induced by stimulation with pro-inflammatory cytokines. Induction was not inhibited by sulfasalazine or gliotoxine, whereas dexamethasone further enhanced both gene transcription and antimicrobial capacity. The lack of inhibition of induced hBD-2 expression by specific NF-kappaB antagonists suggests an additional pathway of activation, independent of NF-kappaB. The induction of hBD-2 expression in cytokine-stimulated CaCo2 cells by corticosteroids indicates further immunomodulatory ability of steroid hormones not yet understood.
Witthöft
T
T
Department of Medicine I, Division of Gastroenterology, University Hospital of Schleswig-Holstein, Campus Lübeck, Lübeck, Germany.
Pilz
C S
CS
Fellermann
K
K
Nitschke
M
M
Stange
E F
EF
Ludwig
D
D
D000305:Adrenal Cortex Hormones / Q000494:pharmacology*; D018938:Caco-2 Cells; D003106:Colon / Q000378:metabolism*; D015169:Colony Count, Microbial; D016207:Cytokines / Q000494:pharmacology; D004847:Epithelial Cells / Q000378:metabolism; D004926:Escherichia coli / Q000254:growth & development; D005912:Gliotoxin / Q000494:pharmacology; D006801:Humans; D016209:Interleukin-8 / Q000235:genetics / Q000378:metabolism; D016328:NF-kappa B / Q000037:antagonists & inhibitors / Q000378:metabolism*; D012333:RNA, Messenger / Q000378:metabolism; D012460:Sulfasalazine / Q000494:pharmacology; D023083:beta-Defensins / Q000235:genetics / Q000378:metabolism* / Q000502:physiology
D000305:Adrenal Cortex Hormones; D016207:Cytokines; C406407:DEFB1 protein, human; C106641:DEFB4A protein, human; D016209:Interleukin-8; D016328:NF-kappa B; D012333:RNA, Messenger; D023083:beta-Defensins; D012460:Sulfasalazine; D005912:Gliotoxin