Nod1 responds to peptidoglycan delivered by the Helicobacter pylori cag pathogenicity island

Epithelial cells can respond to conserved bacterial products that are internalized after either bacterial invasion or liposome treatment of cells. We report here that the noninvasive Gram-negative pathogen Helicobacter pylori was recognized by epithelial cells via Nod1, an intracellular pathogen-recognition molecule with specificity for Gram-negative peptidoglycan. Nod1 detection of H. pylori depended on the delivery of peptidoglycan to host cells by a bacterial type IV secretion system, encoded by the H. pylori cag pathogenicity island. Consistent with involvement of Nod1 in host defense, Nod1-deficient mice were more susceptible to infection by cag pathogenicity island–positive H. pylori than were wild-type mice. We propose that sensing of H. pylori by Nod1 represents a model for host recognition of noninvasive pathogens.

[1]  T. Meyer,et al.  Functional Analysis of the cag Pathogenicity Island in Helicobacter pylori Isolates from Patients with Gastritis, Peptic Ulcer, and Gastric Cancer , 2004, Infection and Immunity.

[2]  R. Peek,et al.  Host and microbial constituents influence Helicobacter pylori-induced cancer in a murine model of hypergastrinemia. , 2003, Gastroenterology.

[3]  A. Labigne,et al.  Essential role of Helicobacter pyloriγ‐glutamyltranspeptidase for the colonization of the gastric mucosa of mice , 1999, Molecular microbiology.

[4]  J. Bertin,et al.  Human CARD4 Protein Is a Novel CED-4/Apaf-1 Cell Death Family Member That Activates NF-κB* , 1999, The Journal of Biological Chemistry.

[5]  Y. Shiratori,et al.  Distinct Mechanism of Helicobacter pylori-mediated NF-κB Activation between Gastric Cancer Cells and Monocytic Cells* , 2001, The Journal of Biological Chemistry.

[6]  P. Laniece,et al.  A new high resolution radioimager for the quantitative analysis of radiolabelled molecules in tissue section , 1998, Journal of Neuroscience Methods.

[7]  M. Kagnoff,et al.  Nod1 Is an Essential Signal Transducer in Intestinal Epithelial Cells Infected with Bacteria That Avoid Recognition by Toll-Like Receptors , 2004, Infection and Immunity.

[8]  M. Borodovsky,et al.  cag, a pathogenicity island of Helicobacter pylori, encodes type I-specific and disease-associated virulence factors. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[9]  H. D. Reuse,et al.  The Helicobacter pylori UreI protein is not involved in urease activity but is essential for bacterial survival in vivo. , 1998, Infection and immunity.

[10]  F. Bäckhed,et al.  Toll-like receptor 4-mediated signaling by epithelial surfaces: necessity or threat? , 2003, Microbes and infection.

[11]  J. Han,et al.  NF-kappaB and c-Jun-dependent regulation of macrophage inflammatory protein-2 gene expression in response to lipopolysaccharide in RAW 264.7 cells. , 2003, Molecular immunology.

[12]  R. Peek,et al.  Helicobacter pylori flagellin evades toll-like receptor 5-mediated innate immunity. , 2004, The Journal of infectious diseases.

[13]  S. Foster,et al.  An essential role for NOD1 in host recognition of bacterial peptidoglycan containing diaminopimelic acid , 2003, Nature Immunology.

[14]  A. Labigne,et al.  Immune Responses of Specific-Pathogen-Free Mice to Chronic Helicobacter pylori (Strain SS1) Infection , 1998, Infection and Immunity.

[15]  R. Rappuoli,et al.  Helicobacter pylori: molecular evolution of a bacterial quasi-species. , 1998, Current opinion in microbiology.

[16]  S. Saccani,et al.  Modulation of NF-κB Activity by Exchange of Dimers , 2003 .

[17]  M. Blaser,et al.  Helicobacter pylori strain-specific differences in genetic content, identified by microarray, influence host inflammatory responses. , 2001, The Journal of clinical investigation.

[18]  B. Glauner Separation and quantification of muropeptides with high-performance liquid chromatography. , 1988, Analytical biochemistry.

[19]  D. Louvard,et al.  5 Epithelial cells: Establishment of primary cultures and immortalization , 2002 .

[20]  S. Falkow,et al.  Helicobacter pylori enter and survive within multivesicular vacuoles of epithelial cells , 2002, Cellular microbiology.

[21]  C. Josenhans,et al.  Helicobacter pylori flagellins have very low intrinsic activity to stimulate human gastric epithelial cells via TLR5. , 2003, Microbes and infection.

[22]  Mark Borodovsky,et al.  The complete genome sequence of the gastric pathogen Helicobacter pylori , 1997, Nature.

[23]  B. Roe,et al.  Analyses of the cag pathogenicity island of Helicobacter pylori , 1998, Molecular microbiology.

[24]  J. Goldberg,et al.  Toll-like Receptor (TLR) 2 and TLR5, but Not TLR4, Are Required for Helicobacter pylori-induced NF-κB Activation and Chemokine Expression by Epithelial Cells* , 2003, Journal of Biological Chemistry.

[25]  R. Haas,et al.  CagA tyrosine phosphorylation and interleukin-8 induction by Helicobacter pylori are independent from alpAB, HopZ and bab group outer membrane proteins. , 2002, International journal of medical microbiology : IJMM.

[26]  J. Bertin,et al.  Nod1 Detects a Unique Muropeptide from Gram-Negative Bacterial Peptidoglycan , 2003, Science.

[27]  E. Cascales,et al.  The versatile bacterial type IV secretion systems , 2003, Nature Reviews Microbiology.

[28]  G. Sherlock,et al.  A whole-genome microarray reveals genetic diversity among Helicobacter pylori strains. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[29]  R. Haas,et al.  Systematic mutagenesis of the Helicobacter pylori cag pathogenicity island: essential genes for CagA translocation in host cells and induction of interleukin‐8 , 2003, Molecular microbiology.

[30]  A. Petersen,et al.  Helicobacter pylori: an invading microorganism? A review. , 2003, FEMS immunology and medical microbiology.

[31]  L. del Peso,et al.  Nod1, an Apaf-1-like Activator of Caspase-9 and Nuclear Factor-κB* , 1999, The Journal of Biological Chemistry.

[32]  M. Blaser,et al.  Heightened inflammatory response and cytokine expression in vivo to cagA+ Helicobacter pylori strains. , 1995, Laboratory investigation; a journal of technical methods and pathology.

[33]  M. Chamaillard,et al.  Nod2 Is a General Sensor of Peptidoglycan through Muramyl Dipeptide (MDP) Detection* , 2003, The Journal of Biological Chemistry.

[34]  I. Helander,et al.  Compositional analysis of Helicobacter pylori rough-form lipopolysaccharides , 1992, Journal of bacteriology.

[35]  S. Chen,et al.  Nod1, an Apaf-1-like activator of caspase-9 and nuclear factor-kappaB. , 1999, The Journal of biological chemistry.

[36]  M. Blaser,et al.  Plasticity of Repetitive DNA Sequences within a Bacterial (Type IV) Secretion System Component , 2003, The Journal of experimental medicine.

[37]  D. Heuermann,et al.  A stable shuttle vector system for efficient genetic complementation of Helicobacter pylori strains by transformation and conjugation , 1998, Molecular and General Genetics MGG.

[38]  A. Lee,et al.  Atrophic gastric changes in both Helicobacter felis and Helicobacter pylori infected mice are host dependent and separate from antral gastritis. , 1996, Gut.

[39]  A. Baldwin,et al.  Activation of Nuclear Factor-κB-dependent Transcription by Tumor Necrosis Factor-α Is Mediated through Phosphorylation of RelA/p65 on Serine 529* , 1998, The Journal of Biological Chemistry.

[40]  M. Selbach,et al.  The Helicobacter pylori CagA protein induces cortactin dephosphorylation and actin rearrangement by c‐Src inactivation , 2003, The EMBO journal.

[41]  F. Bäckhed,et al.  Gastric mucosal recognition of Helicobacter pylori is independent of Toll-like receptor 4. , 2003, The Journal of infectious diseases.

[42]  A. Labigne,et al.  The GroES homolog of Helicobacter pylori confers protective immunity against mucosal infection in mice. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[43]  S. Czinn,et al.  Vaccine-Induced Reduction of Helicobacter pylori Colonization in Mice Is Interleukin-12 Dependent but Gamma Interferon and Inducible Nitric Oxide Synthase Independent , 2003, Infection and Immunity.

[44]  S. Saccani,et al.  Modulation of NF-kappaB activity by exchange of dimers. , 2003, Molecular cell.

[45]  D. Philpott,et al.  Reduced activation of inflammatory responses in host cells by mouse‐adapted Helicobacter pylori isolates , 2002, Cellular microbiology.

[46]  M. Blaser,et al.  Infection with Helicobacter pylori strains possessing cagA is associated with an increased risk of developing adenocarcinoma of the stomach. , 1995, Cancer research.

[47]  A. Cerami,et al.  Genomic cloning and promoter analysis of macrophage inflammatory protein (MIP)-2, MIP-1 alpha, and MIP-1 beta, members of the chemokine superfamily of proinflammatory cytokines. , 1993, Journal of immunology.

[48]  J. Parsonnet Helicobacter pylori: the size of the problem , 1998, Gut.

[49]  J. Han,et al.  NF-κB and c-Jun-dependent regulation of macrophage inflammatory protein-2 gene expression in response to lipopolysaccharide in RAW 264.7 cells , 2003 .

[50]  R. Haas,et al.  Translocation of Helicobacter pylori CagA into gastric epithelial cells by type IV secretion. , 2000, Science.

[51]  J. Bertin,et al.  CARD4/Nod1 mediates NF-kappaB and JNK activation by invasive Shigella flexneri. , 2001, EMBO reports.

[52]  M. Rohde,et al.  A novel sheathed surface organelle of the Helicobacter pylori cag type IV secretion system , 2003, Molecular microbiology.

[53]  S. Garg,et al.  CXC chemokine redundancy ensures local neutrophil recruitment during acute inflammation. , 2001, The American journal of pathology.

[54]  J. Bertin,et al.  CARD4/Nod1 mediates NF‐κB and JNK activation by invasive Shigella flexneri , 2001 .

[55]  D. Wang,et al.  Activation of nuclear factor-kappaB-dependent transcription by tumor necrosis factor-alpha is mediated through phosphorylation of RelA/p65 on serine 529. , 1998, The Journal of biological chemistry.

[56]  R. Haas,et al.  Systematic mutagenesis of the Helicobacter pylori cag pathogenicity island: essential genes for CagA translocation in host cells and induction of interleukin‐8 , 2001 .