CARD15/NOD2 Is Required for Peyer's Patches Homeostasis in Mice

PLOS ONE, Jun 2007

Background CARD15/NOD2 mutations are associated with susceptibility to Crohn's Disease (CD) and Graft Versus Host Disease (GVHD). CD and GVHD are suspected to be related with the dysfunction of Peyer's patches (PP) and isolated lymphoid follicles (LFs). Using a new mouse model invalidated for Card15/Nod2 (KO), we thus analysed the impact of the gene in these lymphoid formations together with the development of experimental colitis. Methodology/Principal Findings At weeks 4, 12 and 52, the numbers of PPs and LFs were higher in KO mice while no difference was observed at birth. At weeks 4 and 12, the size and cellular composition of PPs were analysed by flow cytometry and immunohistochemistry. PPs of KO mice were larger with an increased proportion of M cells and CD4+ T-cells. KO mice were also characterised by higher concentrations of TNFα, IFNγ, IL12 and IL4 measured by ELISA. In contrast, little differences were found in the PP-free ileum and the spleen of KO mice. By Ussing chamber experiments, we found that this PP phenotype is associated with an increased of both paracellular permeability and yeast/bacterial translocation. Finally, KO mice were more susceptible to the colitis induced by TNBS. Conclusions Card15/Nod2 deficiency induces an abnormal development and function of the PPs characterised by an exaggerated immune response and an increased permeability. These observations provide a comprehensive link between the molecular defect and the Human CARD15/NOD2 associated disorders: CD and GVHD.

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CARD15/NOD2 Is Required for Peyer's Patches Homeostasis in Mice

et al (2007) CARD15/NOD2 Is Required for Peyer's Patches Homeostasis in Mice. PLoS ONE 2(6): e523. doi:10.1371/journal.pone.0000523 CARD15/NOD2 Is Required for Peyer's Patches Homeostasis in Mice Fre d erick Barreau 0 1 Ulrich Meinzer 0 1 Fabrice Chareyre 0 1 Dominique Berrebi 0 1 Michiko Niwa-Kawakita 0 1 Monique Dussaillant 0 1 Benoit Foligne 0 1 Vincent Ollendorff 0 1 Martine Heyman 0 1 Ste phane Bonacorsi 0 1 Thecla Lesuffleur 0 1 Ghislaine Sterkers 0 1 Marco Giovannini 0 1 Jean-Pierre Hugot 0 1 0 Academic Editor: Jacques Zimmer, Centre de Recherche Public-Sante , Luxembourg 1 1 U843, INSERM, Paris, France , 2 UMR-S843 , Universite Paris Diderot , Paris , France , 3 Service de Gastroente rologie, H opital R. Debre , AP-HP, Paris, France, 4 Institut Universitaire d'H e matologie, Universite Paris Diderot , Paris, France, 5 U674, INSERM, Paris, France, 6 EA3102 , Universite Paris Diderot , Paris , France , 7 Service d'Anatomie Pathologique, Institut Pasteur de Lille, Lille, France, 8 Laboratoire des Bacte ries Lactiques et Immunite des Muqueuses, Institut Pasteur de Lille, Lille, France, 9 IMRN and UMR 1111 INRA, Facult e Saint-Je ro me, Universite Paul Ce zanne , Marseille, France, 10 U793, INSERM, Paris, France, 11 IFR94 , Universite Paris Descartes, Paris, France , 12 EA3105 , Universite Paris Diderot , Paris , France , 13 Service de Microbiologie, H opital R. Debre , AP-HP, Paris, France, 14 Service d'Immunologie, H opital R. Debre , AP-HP , Paris , France Background. CARD15/NOD2 mutations are associated with susceptibility to Crohn's Disease (CD) and Graft Versus Host Disease (GVHD). CD and GVHD are suspected to be related with the dysfunction of Peyer's patches (PP) and isolated lymphoid follicles (LFs). Using a new mouse model invalidated for Card15/Nod2 (KO), we thus analysed the impact of the gene in these lymphoid formations together with the development of experimental colitis. Methodology/Principal Findings. At weeks 4, 12 and 52, the numbers of PPs and LFs were higher in KO mice while no difference was observed at birth. At weeks 4 and 12, the size and cellular composition of PPs were analysed by flow cytometry and immunohistochemistry. PPs of KO mice were larger with an increased proportion of M cells and CD4+ T-cells. KO mice were also characterised by higher concentrations of TNFa, IFNc, IL12 and IL4 measured by ELISA. In contrast, little differences were found in the PP-free ileum and the spleen of KO mice. By Ussing chamber experiments, we found that this PP phenotype is associated with an increased of both paracellular permeability and yeast/bacterial translocation. Finally, KO mice were more susceptible to the colitis induced by TNBS. Conclusions. Card15/Nod2 deficiency induces an abnormal development and function of the PPs characterised by an exaggerated immune response and an increased permeability. These observations provide a comprehensive link between the molecular defect and the Human CARD15/NOD2 associated disorders: CD and GVHD. - INTRODUCTION Caspase Recruitment Domain 15 (CARD15) also known as Nucleotide oligomerisation domain 2 (NOD2) has been associated with Crohns Disease (CD) [1,2] and graft versus host disease (GVHD) [3,4]. NOD2 belongs to a family of genes involved in innate immunity [5]. It can be activated by muropeptides which are components of the bacterial cell wall. When activated, NOD2 interacts with Rick/Rip2 which in turn activates the NF-kB pathway, resulting in the production of pro-inflammatory cytokines. Half of CD patients have one or more NOD2 mutations [6]. Some of the CD associated mutations were found unresponsive to muropeptides [5]. By consequence, CD is usually considered as an immune deficiency toward bacteria present in the gut lumen [7]. However, the exact mechanism by which NOD2 mutations are able to induce CD lesions is still subject to debate [710]. Holler et al. reported that the three major mutations associated with CD (R702W, G908R and 1007fs) are also associated with severe acute GVHD and bone marrow transplantation (BMT) related mortality [3]. Mutations in both donor and recipient were found deleterious, suggesting a role of epithelial and circulating cells in disease mechanisms. Despite some differences in their conclusions, other groups recently confirmed the association between NOD2 and BMT complications [4,11]. CD is a chronic relapsing inflammatory bowel disease (IBD) with mucosal ulcerations of the digestive tract. CD lesions are characterised by a T helper (Th) 1 immune response and several authors have shown that they are related with gut associated lymphoid tissue (GALT), known as lymphoid follicles (LFs). LFs are mainly encountered in the colon where they are isolated and in small bowel where they are grouped forming Peyers patches (PP) which are known to be pivotal sites for the host immune response and for the entry of enteropathogen bacteria. CD lesions are most often localized in colon and distal ileu (...truncated)


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Frédérick Barreau, Ulrich Meinzer, Fabrice Chareyre, Dominique Berrebi, Michiko Niwa-Kawakita, Monique Dussaillant, Benoit Foligne, Vincent Ollendorff, Martine Heyman, Stéphane Bonacorsi, Thecla Lesuffleur, Ghislaine Sterkers, Marco Giovannini, Jean-Pierre Hugot. CARD15/NOD2 Is Required for Peyer's Patches Homeostasis in Mice, PLOS ONE, 2007, 6, DOI: 10.1371/journal.pone.0000523