Bacteroides fragilis Enterotoxin Induces Intestinal Epithelial Cell Secretion of Interleukin-8 through Mitogen-Activated Protein Kinases and a Tyrosine Kinase-Regulated Nuclear Factor-κB Pathway

ABSTRACT Enterotoxigenic Bacteroides fragilis (ETBF) secretes a 20-kDa metalloprotease toxin termed B. fragilis toxin (BFT). ETBF disease in animals is associated with an acute inflammatory response in the intestinal mucosa, and lethal hemorrhagic colitis may occur in rabbits. In this study, we confirmed recent reports (J. M. Kim, Y. K. Oh, Y. J. Kim, H. B. Oh, and Y. J. Cho, Clin. Exp. Immunol. 123:421-427, 2001; L. Sanfilippo, C. K. Li, R. Seth, T. J. Balwin, M. J. Menozzi, and Y. R. Mahida, Clin. Exp. Immunol. 119:456-463, 2000) that purified BFT stimulates interleukin-8 (IL-8) secretion by human intestinal epithelial cells (HT29/C1 cells) and demonstrate that stimulation of IL-8 production is dependent on biologically active BFT and independent of serum. Induction of IL-8 mRNA expression occurs rapidly and ceases by 6 h after BFT treatment, whereas IL-8 secretion continues to increase for at least 18 h. Our data suggest that BFT-stimulated IL-8 secretion involves tyrosine kinase-dependent activation of nuclear factor-κB (NF-κB) as well as activation of the mitogen-activated protein kinases (MAPKs), p38 and extracellular signal-related kinase. Simultaneous activation of NF-κB and MAPKs appears necessary for secretion of IL-8 by HT29/C1 cells treated with BFT.

[1]  C. Lowenstein,et al.  Inducible nitric oxide synthase expression inhibition by adenovirus E1A , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[2]  C. Lau,et al.  Title Differential Effects of Tyrosine Kinase Inhibitors on Volume-sensitive Chloride Current in Human Atrial Myocytes : Evidence for Dual Regulation by Src and EGFR Kinases , 2004 .

[3]  T. Wilkins,et al.  The Bacteroides fragilis toxin fragilysin disrupts the paracellular barrier of epithelial cells , 1997, Infection and immunity.

[4]  Y. Oh,et al.  Polarized secretion of CXC chemokines by human intestinal epithelial cells in response to Bacteroides fragilis enterotoxin: NF‐κB plays a major role in the regulation of IL‐8 expression , 2001, Clinical and experimental immunology.

[5]  D. Acheson,et al.  Shiga Toxins Stimulate Secretion of Interleukin-8 from Intestinal Epithelial Cells , 1999, Infection and Immunity.

[6]  W. Moore,et al.  Human fecal flora: the normal flora of 20 Japanese-Hawaiians. , 1974, Applied microbiology.

[7]  A. Baird,et al.  Tissue cytokine and chemokine expression in inflammatory bowel disease , 2001, Inflammation Research.

[8]  S. Savkovic,et al.  EPEC-activated ERK1/2 participate in inflammatory response but not tight junction barrier disruption. , 2001, American journal of physiology. Gastrointestinal and liver physiology.

[9]  M. Kagnoff,et al.  A distinct array of proinflammatory cytokines is expressed in human colon epithelial cells in response to bacterial invasion. , 1995, The Journal of clinical investigation.

[10]  L. Mahadevan,et al.  Transcription: MAPK-regulated transcription: a continuously variable gene switch? , 2002, Nature Reviews Molecular Cell Biology.

[11]  J. Silva,et al.  Bacteroides fragilis enterotoxin gene sequences in patients with inflammatory bowel disease. , 2000, Emerging infectious diseases.

[12]  D. Thuerauf,et al.  p38 MAPK and NF-kappa B collaborate to induce interleukin-6 gene expression and release. Evidence for a cytoprotective autocrine signaling pathway in a cardiac myocyte model system. , 2000, The Journal of biological chemistry.

[13]  R. Sack,et al.  Human colonic epithelial cells, HT29/C1, treated with crude Bacteroides fragilis enterotoxin dramatically alter their morphology , 1992, Infection and immunity.

[14]  R. Sartor,et al.  The I kappa B/NF-kappa B system: a key determinant of mucosalinflammation and protection. , 2000, American journal of physiology. Cell physiology.

[15]  C. Sears,et al.  Bacteroides fragilis enterotoxin cleaves the zonula adherens protein, E-cadherin. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[16]  J. Sambrook,et al.  Molecular Cloning: A Laboratory Manual , 2001 .

[17]  F. Herrmann,et al.  Control of transcriptional activation of the lipopolysaccharide binding protein (LBP) gene by proinflammatory cytokines. , 1997, Cytokines, cellular & molecular therapy.

[18]  J. Swantek,et al.  New insights into the control of MAP kinase pathways. , 1999, Experimental Cell Research.

[19]  Y. Mahida,et al.  NF‐κB may determine whether epithelial cell–microbial interactions in the intestine are hostile or friendly , 2001, Clinical and experimental immunology.

[20]  C. Lau,et al.  Differential Effects of Tyrosine Kinase Inhibitors on Volume-sensitive Chloride Current in Human Atrial Myocytes , 2004, The Journal of general physiology.

[21]  M. Laburthe,et al.  Protease-activated Receptor 2 in Colon Cancer , 2004, Journal of Biological Chemistry.

[22]  Jerry L. Adams,et al.  A protein kinase involved in the regulation of inflammatory cytokine biosynthesis , 1994, Nature.

[23]  J. Collins,et al.  Rabbit model to evaluate enterovirulence of Bacteroides fragilis , 1990, Journal of clinical microbiology.

[24]  Cynthia L Sears,et al.  Bacteroides fragilis enterotoxin induces c-Myc expression and cellular proliferation. , 2001, Gastroenterology.

[25]  Penny A. Johnson,et al.  Chemokine expression in IBD. Mucosal chemokine expression is unselectively increased in both ulcerative colitis and Crohn's disease , 2003, The Journal of pathology.

[26]  T. Maniatis,et al.  NF-kappa B: a lesson in family values. , 1995, Cell.

[27]  C. Pothoulakis,et al.  p38 MAP kinase activation by Clostridium difficile toxin A mediates monocyte necrosis, IL-8 production, and enteritis. , 2000, The Journal of clinical investigation.

[28]  A. Tzianabos,et al.  Diversity of the Metalloprotease Toxin Produced by Enterotoxigenic Bacteroides fragilis , 2002, Infection and Immunity.

[29]  T. Maniatis,et al.  NF-κB: A lesson in family values , 1995, Cell.

[30]  R. Winzen,et al.  Stress-activated Protein Kinase/Jun N-terminal Kinase Is Required for Interleukin (IL)-1-induced IL-6 and IL-8 Gene Expression in the Human Epidermal Carcinoma Cell Line KB* , 1998, The Journal of Biological Chemistry.

[31]  M. Menozzi,et al.  Bacteroides fragilis enterotoxin induces the expression of IL‐8 and transforming growth factor‐beta (TGF‐β) by human colonic epithelial cells , 2000, Clinical and experimental immunology.

[32]  C. Sears,et al.  Detection of toxin production by Bacteroides fragilis: assay development and screening of extraintestinal clinical isolates. , 1996, Clinical infectious diseases : an official publication of the Infectious Diseases Society of America.

[33]  S. Haskill,et al.  Evidence for altered regulation of I kappa B alpha degradation in human colonic epithelial cells. , 1997, Journal of immunology.

[34]  R. Ulevitch,et al.  Lipopolysaccharide activation of human endothelial and epithelial cells is mediated by lipopolysaccharide-binding protein and soluble CD14. , 1993, Proceedings of the National Academy of Sciences of the United States of America.

[35]  J. Collins,et al.  Diarrheal disease caused by enterotoxigenic Bacteroides fragilis in infant rabbits , 1989, Journal of clinical microbiology.

[36]  R. Sack,et al.  Isolation of enterotoxigenic Bacteroides fragilis from humans with diarrhea , 1987, Journal of clinical microbiology.

[37]  A. Pantosti,et al.  Detection of enterotoxigenic Bacteroides fragilis and its toxin in stool samples from adults and children in Italy. , 1997, Clinical infectious diseases : an official publication of the Infectious Diseases Society of America.

[38]  T. Wilkins,et al.  Purification and characterization of an enterotoxin from Bacteroides fragilis , 1992, Infection and immunity.

[39]  D. Clark,et al.  Bacteroides fragilis toxin exhibits polar activity on monolayers of human intestinal epithelial cells (T84 cells) in vitro , 1997, Infection and immunity.

[40]  T. Wilkins,et al.  Proteolytic activity of the Bacteroides fragilis enterotoxin causes fluid secretion and intestinal damage in vivo , 1995, Infection and immunity.

[41]  C. L. Sears,et al.  Enterotoxigenic Bacteroides fragilis. , 1995, Clinical infectious diseases : an official publication of the Infectious Diseases Society of America.

[42]  C. Sears,et al.  The toxins of Bacteroides fragilis. , 2001, Toxicon : official journal of the International Society on Toxinology.

[43]  A. Pantosti,et al.  Detection of Bacteroides fragilis enterotoxin in the feces of a child with diarrhea. , 1994, Clinical infectious diseases : an official publication of the Infectious Diseases Society of America.

[44]  D. Bobak,et al.  Roles of intracellular calcium and NF-kappa B in the Clostridium difficile toxin A-induced up-regulation and secretion of IL-8 from human monocytes. , 1999, Journal of immunology.

[45]  Pascale Jeannin,et al.  Measurement of nuclear factor-kappa B translocation on lipopolysaccharide-activated human dendritic cells by confocal microscopy and flow cytometry. , 2002, Cytometry.

[46]  D. S. Shoop,et al.  Association of Enterotoxigenic Bacteroides fragilis with Diarrheal Disease in Calves , 1985, The Journal of infectious diseases.

[47]  T. Murayama,et al.  [Molecular mechanism of interleukin-8 gene expression]. , 1994, Rinsho byori. The Japanese journal of clinical pathology.

[48]  C. Pothoulakis,et al.  Bacteroides fragilis toxin 2 damages human colonic mucosa in vitro , 1999, Gut.

[49]  R. Sartor,et al.  The IκB/NF-κB system: a key determinant of mucosal inflammation and protection , 2000 .

[50]  J. Brazier,et al.  Medical and Environmental Aspects of Anaerobes , 1992 .