Human β-defensin 2 but not β-defensin 1 is expressed preferentially in colonic mucosa of inflammatory bowel disease

Objective Various antimicrobial peptides such as defensins are part of innate immunity and contribute to the intestinal barrier that may be defective in inflammatory bowel disease (IBD). This study investigated β-defensin mRNA and peptide expression in the colon from controls and patients with Crohn's disease, ulcerative colitis or unspecific colitis as inflammatory controls. Methods Mucosal mRNA expression was measured by multiplex reverse transcriptase–polymerase chain reaction (RT-PCR) with primers for human β-defensin 1 (HBD-1) and human β-defensin 2 (HBD-2) in CaCo-2 cells and in biopsies from 103 patients (33 controls, 24 Crohn's disease patients, 36 ulcerative colitis patients, 10 unspecific colitis patients). Paraffin-embedded tissue from colonic resections was tested for HBD-1 and HBD-2 peptides by immunohistochemistry. Results HBD-1 mRNA was expressed constitutively whereas HBD-2 was induced by pro-inflammatory cytokines in CaCo-2 cells. HBD-1 mRNA was detectable in 61% of control and Crohn's disease biopsies and 53% of ulcerative colitis biopsies. HBD-2 transcript was expressed differentially, with 18% of control biopsies positive as opposed to 34% in Crohn's disease and 53% in ulcerative colitis. HBD-2 mRNA but not HBD-1 mRNA was expressed preferentially in inflamed areas. Immunohistochemical investigation demonstrated the presence of defensin peptides in colonic epithelium as well as the differential induction in IBD. Conclusions HBD-1 is expressed constitutively in colonic tissue irrespective of inflammation. HBD-2 is barely present in uninflamed colon but it is induced in inflammation. The lower expression of HBD-2 in Crohn's disease compared with ulcerative colitis indicates different responses of the mucosal innate defence. Defensins may play a crucial role in controlling pathogen invasion in IBD, although the functional significance remains to be established.

[1]  K. Fellermann,et al.  Defensins – innate immunity at the epithelial frontier , 2001, European journal of gastroenterology & hepatology.

[2]  Mourad Sahbatou,et al.  Association of NOD2 leucine-rich repeat variants with susceptibility to Crohn's disease , 2001, Nature.

[3]  Judy H. Cho,et al.  A frameshift mutation in NOD2 associated with susceptibility to Crohn's disease , 2001, Nature.

[4]  J. Schröder,et al.  Isolation and Characterization of Human β-Defensin-3, a Novel Human Inducible Peptide Antibiotic* , 2001, The Journal of Biological Chemistry.

[5]  W. Chan,et al.  Human defensin 5 is stored in precursor form in normal Paneth cells and is expressed by some villous epithelial cells and by metaplastic Paneth cells in the colon in inflammatory bowel disease , 2001, Gut.

[6]  M. Kagnoff,et al.  Expression and regulation of the human beta-defensins hBD-1 and hBD-2 in intestinal epithelium. , 1999, Journal of immunology.

[7]  K. Fellermann,et al.  Enhanced Intestinal Expression of Heat Shock Protein 70 in Patients with Inflammatory Bowel Diseases , 1999, Digestive Diseases and Sciences.

[8]  M. Neurath,et al.  T cell specificity and cross reactivity towards enterobacteria,Bacteroides,Bifidobacterium, and antigens from resident intestinal flora in humans , 1999, Gut.

[9]  A. Kruse,et al.  Mucosal cytokine expression, cellular markers and adhesion molecules in inflammatory bowel disease. , 1999, European journal of gastroenterology & hepatology.

[10]  E. Greenberg,et al.  Production of β-defensins by human airway epithelia , 1998 .

[11]  N. Barnich,et al.  Presence of adherent Escherichia coli strains in ileal mucosa of patients with Crohn's disease. , 1998, Gastroenterology.

[12]  H. Heng,et al.  Structure and mapping of the human beta-defensin HBD-2 gene and its expression at sites of inflammation. , 1998, Gene.

[13]  T. Ganz,et al.  Isolation of human intestinal defensins from ileal neobladder urine , 1998, FEBS letters.

[14]  S. Krisanaprakornkit,et al.  Expression of the Peptide Antibiotic Human β-Defensin 1 in Cultured Gingival Epithelial Cells and Gingival Tissue , 1998, Infection and Immunity.

[15]  V. Bafna,et al.  Human beta-defensin 2 is a salt-sensitive peptide antibiotic expressed in human lung. , 1998, The Journal of clinical investigation.

[16]  M. Nakazato,et al.  Identification of human beta-defensin-2 in respiratory tract and plasma and its increase in bacterial pneumonia. , 1998, Biochemical and biophysical research communications.

[17]  R. Knuechel,et al.  Nuclear factor kappaB is activated in macrophages and epithelial cells of inflamed intestinal mucosa. , 1998, Gastroenterology.

[18]  K. Fellermann,et al.  Intestinal Expression of Human Heat Shock Protein 90 in Patients with Crohn's Disease and Ulcerative Colitis , 1998, Digestive Diseases and Sciences.

[19]  Christina H. Park,et al.  Human beta-defensin-1: an antimicrobial peptide of urogenital tissues. , 1998, The Journal of clinical investigation.

[20]  J. Hampe,et al.  Activation of nuclear factor κB in inflammatory bowel disease , 1998, Gut.

[21]  Douglas R. Morgan,et al.  Sleisenger and Fordtran's Gastrointestinal and Liver Disease: Pathophysiology/Diagnosis/Management , 1998 .

[22]  Douglas E. Jones,et al.  Enteric β-Defensin: Molecular Cloning and Characterization of a Gene with Inducible Intestinal Epithelial Cell Expression Associated with Cryptosporidium parvumInfection , 1998, Infection and Immunity.

[23]  P. Rutgeerts,et al.  Early lesions of recurrent Crohn's disease caused by infusion of intestinal contents in excluded ileum. , 1998, Gastroenterology.

[24]  R. Siebert,et al.  Mapping of the gene encoding human beta-defensin-2 (DEFB2) to chromosome region 8p22-p23.1. , 1997, Genomics.

[25]  H. Heng,et al.  The human β-defensin-1 and α-defensins are encoded by adjacent genes : Two peptide families with differing disulfide topology share a common ancestry , 1997 .

[26]  J. Schröder,et al.  A peptide antibiotic from human skin , 1997, Nature.

[27]  James M. Wilson,et al.  Human β-Defensin-1 Is a Salt-Sensitive Antibiotic in Lung That Is Inactivated in Cystic Fibrosis , 1997, Cell.

[28]  R. Lehrer,et al.  Widespread expression of beta‐defensin hBD‐1 in human secretory glands and epithelial cells , 1996, FEBS letters.

[29]  J. Reimund,et al.  Increased production of tumour necrosis factor-alpha interleukin-1 beta, and interleukin-6 by morphologically normal intestinal biopsies from patients with Crohn's disease. , 1996, Gut.

[30]  G. Diamond,et al.  Inducible expression of an antibiotic peptide gene in lipopolysaccharide-challenged tracheal epithelial cells. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[31]  E. Greenberg,et al.  Cystic Fibrosis Airway Epithelia Fail to Kill Bacteria Because of Abnormal Airway Surface Fluid , 1996, Cell.

[32]  C. Prantera,et al.  An antibiotic regimen for the treatment of active Crohn's disease: a randomized, controlled clinical trial of metronidazole plus ciprofloxacin. , 1996, The American journal of gastroenterology.

[33]  B. Schonwetter,et al.  Epithelial antibiotics induced at sites of inflammation. , 1995, Science.

[34]  P. Rutgeerts,et al.  Controlled trial of metronidazole treatment for prevention of Crohn's recurrence after ileal resection. , 1995, Gastroenterology.

[35]  M. Selsted,et al.  Mouse Paneth cell defensins: primary structures and antibacterial activities of numerous cryptdin isoforms , 1994, Infection and immunity.

[36]  V. Binder,et al.  Mycobacterium paratuberculosis in intestinal tissue from patients with Crohn's disease demonstrated by a nested primer polymerase chain reaction. , 1994, Scandinavian journal of gastroenterology.

[37]  G. Rook,et al.  Specific detection of Mycobacterium paratuberculosis DNA associated with granulomatous tissue in Crohn's disease. , 1994, Gut.

[38]  S. Tonegawa,et al.  Spontaneous development of inflammatory bowel disease in T cell receptor mutant mice , 1993, Cell.

[39]  A. Feller,et al.  Ulcerative colitis-like disease in mice with a disrupted interleukin-2 gene , 1993, Cell.

[40]  K. Rajewsky,et al.  Interleukin-10-deficient mice develop chronic enterocolitis , 1993, Cell.

[41]  S. Miller,et al.  Enteric defensins: antibiotic peptide components of intestinal host defense , 1992, The Journal of cell biology.

[42]  P. Rutgeerts,et al.  Effect of faecal stream diversion on recurrence of Crohn's disease in the neoterminal ileum , 1991, The Lancet.

[43]  M. Zasloff,et al.  Tracheal antimicrobial peptide, a cysteine-rich peptide from mammalian tracheal mucosa: peptide isolation and cloning of a cDNA. , 1991, Proceedings of the National Academy of Sciences of the United States of America.

[44]  T. Ganz,et al.  Interaction of human defensins with Escherichia coli. Mechanism of bactericidal activity. , 1989, The Journal of clinical investigation.

[45]  D. Jewell,et al.  Split ileostomy and ileocolostomy for Crohn's disease of the colon and ulcerative colitis: a 20 year survey. , 1983, Gut.

[46]  R. Colman,et al.  Experimental models of inflammatory bowel disease. , 2003, Archivum immunologiae et therapiae experimentalis.

[47]  E. Greenberg,et al.  Production of beta-defensins by human airway epithelia. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[48]  D. Clark,et al.  Enteric beta-defensin: molecular cloning and characterization of a gene with inducible intestinal epithelial cell expression associated with Cryptosporidium parvum infection. , 1998, Infection and immunity.

[49]  H. Heng,et al.  The human beta-defensin-1 and alpha-defensins are encoded by adjacent genes: two peptide families with differing disulfide topology share a common ancestry. , 1997, Genomics.

[50]  A. Hayday,et al.  T cell receptor-alpha beta-deficient mice fail to develop colitis in the absence of a microbial environment. , 1997, The American journal of pathology.

[51]  A. Dutra,et al.  Human enteric defensin genes: chromosomal map position and a model for possible evolutionary relationships. , 1996, Genomics.

[52]  T. Ganz,et al.  Antimicrobial defensin peptides form voltage-dependent ion-permeable channels in planar lipid bilayer membranes. , 1990, Proceedings of the National Academy of Sciences of the United States of America.