Toll-like receptor 2 regulates intestinal inflammation by controlling integrity of the enteric nervous system

Paola Brun, Maria Cecilia Giron, Marsela Qesari, Andrea Porzionato, Valentina Caputi, Chiara Zoppellaro, Serena Banzato, Alessia Rosaria Grillo, Lisa Spagnol, Raffaele De Caro, Daniela Pizzuti, Vito Barbieri, Antonio Rosato, Giacomo Carlo Sturniolo, Diego Martines, Giovanni Zaninotto, Giorgio Palù, Ignazio Castagliuolo

Research output: Contribution to journalArticle

110 Citations (Scopus)

Abstract

Background & Aims In the intestines, Toll-like receptor 2 (TLR2) mediates immune responses to pathogens and regulates epithelial barrier function; polymorphisms in TLR2 have been associated with inflammatory bowel disease phenotype. We assessed the effects of TLR2 signaling on the enteric nervous system (ENS) in mice. Methods TLR2 distribution and function in the ileal neuromuscular layer of mice were determined by immunofluorescence, cytofluorimetric analysis, immunoprecipitation, and immunoblot analyses. We assessed morphology and function of the ENS in Tlr2-/- mice and in mice with wild-type Tlr2 (wild-type mice) depleted of intestinal microbiota, using immunofluorescence, immunoblot, and gastrointestinal motility assays. Levels and signaling of glial cell line-derived neurotrophic factor (GDNF) were determined using quantitative reverse transcriptase polymerase chain reaction, immunohistochemistry, and immunoprecipitation analyses. Colitis was induced by administration of dextran sulfate sodium or 2,4 dinitrobenzensulfonic acid to Tlr2-/- mice after termination of GDNF administration. Results TLR2 was expressed in enteric neurons, glia, and smooth muscle cells of the intestinal wall. Tlr2-/- mice had alterations in ENS architecture and neurochemical profile, intestinal dysmotility, abnormal mucosal secretion, reduced levels of GDNF in smooth muscle cells, and impaired signaling via Ret-GFRα1. ENS structural and functional anomalies were completely corrected by administration of GDNF to Tlr2-/- mice. Wild-type mice depleted of intestinal microbiota had ENS defects and GDNF deficiency, similar to Tlr2-/- mice; these defects were partially restored by administration of a TLR2 agonist. Tlr2-/- mice developed more severe colitis than wild-type mice after administration of dextran sulfate sodium or 2,4 dinitrobenzensulfonic acid; colitis was not more severe if Tlr2 -/- mice were given GDNF before dextran sulfate sodium or 2,4 dinitrobenzensulfonic acid. Conclusions In mice, TLR2 signaling regulates intestinal inflammation by controlling ENS structure and neurochemical coding, along with intestinal neuromuscular function. These findings provide information as to how defective TLR2 signaling in the ENS affects inflammatory bowel disease phenotype in humans.

Original languageEnglish
Pages (from-to)1323-1333
Number of pages11
JournalGastroenterology
Volume145
Issue number6
DOIs
Publication statusPublished - Dec 2013

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Enteric Nervous System
Toll-Like Receptor 2
Inflammation
Glial Cell Line-Derived Neurotrophic Factor
Dextran Sulfate
Colitis
Inflammatory Bowel Diseases
Immunoprecipitation
Acids
Smooth Muscle Myocytes
Fluorescent Antibody Technique
Phenotype
Gastrointestinal Motility
Reverse Transcriptase Polymerase Chain Reaction
Neuroglia
Cell Wall
Intestines

Keywords

  • Immune Regulation
  • Innate Immunity
  • Microbe
  • Ulcerative Colitis

ASJC Scopus subject areas

  • Gastroenterology

Cite this

Brun, P., Giron, M. C., Qesari, M., Porzionato, A., Caputi, V., Zoppellaro, C., ... Castagliuolo, I. (2013). Toll-like receptor 2 regulates intestinal inflammation by controlling integrity of the enteric nervous system. Gastroenterology, 145(6), 1323-1333. https://doi.org/10.1053/j.gastro.2013.08.047

Toll-like receptor 2 regulates intestinal inflammation by controlling integrity of the enteric nervous system. / Brun, Paola; Giron, Maria Cecilia; Qesari, Marsela; Porzionato, Andrea; Caputi, Valentina; Zoppellaro, Chiara; Banzato, Serena; Grillo, Alessia Rosaria; Spagnol, Lisa; De Caro, Raffaele; Pizzuti, Daniela; Barbieri, Vito; Rosato, Antonio; Sturniolo, Giacomo Carlo; Martines, Diego; Zaninotto, Giovanni; Palù, Giorgio; Castagliuolo, Ignazio.

In: Gastroenterology, Vol. 145, No. 6, 12.2013, p. 1323-1333.

Research output: Contribution to journalArticle

Brun, P, Giron, MC, Qesari, M, Porzionato, A, Caputi, V, Zoppellaro, C, Banzato, S, Grillo, AR, Spagnol, L, De Caro, R, Pizzuti, D, Barbieri, V, Rosato, A, Sturniolo, GC, Martines, D, Zaninotto, G, Palù, G & Castagliuolo, I 2013, 'Toll-like receptor 2 regulates intestinal inflammation by controlling integrity of the enteric nervous system', Gastroenterology, vol. 145, no. 6, pp. 1323-1333. https://doi.org/10.1053/j.gastro.2013.08.047
Brun, Paola ; Giron, Maria Cecilia ; Qesari, Marsela ; Porzionato, Andrea ; Caputi, Valentina ; Zoppellaro, Chiara ; Banzato, Serena ; Grillo, Alessia Rosaria ; Spagnol, Lisa ; De Caro, Raffaele ; Pizzuti, Daniela ; Barbieri, Vito ; Rosato, Antonio ; Sturniolo, Giacomo Carlo ; Martines, Diego ; Zaninotto, Giovanni ; Palù, Giorgio ; Castagliuolo, Ignazio. / Toll-like receptor 2 regulates intestinal inflammation by controlling integrity of the enteric nervous system. In: Gastroenterology. 2013 ; Vol. 145, No. 6. pp. 1323-1333.
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abstract = "Background & Aims In the intestines, Toll-like receptor 2 (TLR2) mediates immune responses to pathogens and regulates epithelial barrier function; polymorphisms in TLR2 have been associated with inflammatory bowel disease phenotype. We assessed the effects of TLR2 signaling on the enteric nervous system (ENS) in mice. Methods TLR2 distribution and function in the ileal neuromuscular layer of mice were determined by immunofluorescence, cytofluorimetric analysis, immunoprecipitation, and immunoblot analyses. We assessed morphology and function of the ENS in Tlr2-/- mice and in mice with wild-type Tlr2 (wild-type mice) depleted of intestinal microbiota, using immunofluorescence, immunoblot, and gastrointestinal motility assays. Levels and signaling of glial cell line-derived neurotrophic factor (GDNF) were determined using quantitative reverse transcriptase polymerase chain reaction, immunohistochemistry, and immunoprecipitation analyses. Colitis was induced by administration of dextran sulfate sodium or 2,4 dinitrobenzensulfonic acid to Tlr2-/- mice after termination of GDNF administration. Results TLR2 was expressed in enteric neurons, glia, and smooth muscle cells of the intestinal wall. Tlr2-/- mice had alterations in ENS architecture and neurochemical profile, intestinal dysmotility, abnormal mucosal secretion, reduced levels of GDNF in smooth muscle cells, and impaired signaling via Ret-GFRα1. ENS structural and functional anomalies were completely corrected by administration of GDNF to Tlr2-/- mice. Wild-type mice depleted of intestinal microbiota had ENS defects and GDNF deficiency, similar to Tlr2-/- mice; these defects were partially restored by administration of a TLR2 agonist. Tlr2-/- mice developed more severe colitis than wild-type mice after administration of dextran sulfate sodium or 2,4 dinitrobenzensulfonic acid; colitis was not more severe if Tlr2 -/- mice were given GDNF before dextran sulfate sodium or 2,4 dinitrobenzensulfonic acid. Conclusions In mice, TLR2 signaling regulates intestinal inflammation by controlling ENS structure and neurochemical coding, along with intestinal neuromuscular function. These findings provide information as to how defective TLR2 signaling in the ENS affects inflammatory bowel disease phenotype in humans.",
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AU - Giron, Maria Cecilia

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AU - Porzionato, Andrea

AU - Caputi, Valentina

AU - Zoppellaro, Chiara

AU - Banzato, Serena

AU - Grillo, Alessia Rosaria

AU - Spagnol, Lisa

AU - De Caro, Raffaele

AU - Pizzuti, Daniela

AU - Barbieri, Vito

AU - Rosato, Antonio

AU - Sturniolo, Giacomo Carlo

AU - Martines, Diego

AU - Zaninotto, Giovanni

AU - Palù, Giorgio

AU - Castagliuolo, Ignazio

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N2 - Background & Aims In the intestines, Toll-like receptor 2 (TLR2) mediates immune responses to pathogens and regulates epithelial barrier function; polymorphisms in TLR2 have been associated with inflammatory bowel disease phenotype. We assessed the effects of TLR2 signaling on the enteric nervous system (ENS) in mice. Methods TLR2 distribution and function in the ileal neuromuscular layer of mice were determined by immunofluorescence, cytofluorimetric analysis, immunoprecipitation, and immunoblot analyses. We assessed morphology and function of the ENS in Tlr2-/- mice and in mice with wild-type Tlr2 (wild-type mice) depleted of intestinal microbiota, using immunofluorescence, immunoblot, and gastrointestinal motility assays. Levels and signaling of glial cell line-derived neurotrophic factor (GDNF) were determined using quantitative reverse transcriptase polymerase chain reaction, immunohistochemistry, and immunoprecipitation analyses. Colitis was induced by administration of dextran sulfate sodium or 2,4 dinitrobenzensulfonic acid to Tlr2-/- mice after termination of GDNF administration. Results TLR2 was expressed in enteric neurons, glia, and smooth muscle cells of the intestinal wall. Tlr2-/- mice had alterations in ENS architecture and neurochemical profile, intestinal dysmotility, abnormal mucosal secretion, reduced levels of GDNF in smooth muscle cells, and impaired signaling via Ret-GFRα1. ENS structural and functional anomalies were completely corrected by administration of GDNF to Tlr2-/- mice. Wild-type mice depleted of intestinal microbiota had ENS defects and GDNF deficiency, similar to Tlr2-/- mice; these defects were partially restored by administration of a TLR2 agonist. Tlr2-/- mice developed more severe colitis than wild-type mice after administration of dextran sulfate sodium or 2,4 dinitrobenzensulfonic acid; colitis was not more severe if Tlr2 -/- mice were given GDNF before dextran sulfate sodium or 2,4 dinitrobenzensulfonic acid. Conclusions In mice, TLR2 signaling regulates intestinal inflammation by controlling ENS structure and neurochemical coding, along with intestinal neuromuscular function. These findings provide information as to how defective TLR2 signaling in the ENS affects inflammatory bowel disease phenotype in humans.

AB - Background & Aims In the intestines, Toll-like receptor 2 (TLR2) mediates immune responses to pathogens and regulates epithelial barrier function; polymorphisms in TLR2 have been associated with inflammatory bowel disease phenotype. We assessed the effects of TLR2 signaling on the enteric nervous system (ENS) in mice. Methods TLR2 distribution and function in the ileal neuromuscular layer of mice were determined by immunofluorescence, cytofluorimetric analysis, immunoprecipitation, and immunoblot analyses. We assessed morphology and function of the ENS in Tlr2-/- mice and in mice with wild-type Tlr2 (wild-type mice) depleted of intestinal microbiota, using immunofluorescence, immunoblot, and gastrointestinal motility assays. Levels and signaling of glial cell line-derived neurotrophic factor (GDNF) were determined using quantitative reverse transcriptase polymerase chain reaction, immunohistochemistry, and immunoprecipitation analyses. Colitis was induced by administration of dextran sulfate sodium or 2,4 dinitrobenzensulfonic acid to Tlr2-/- mice after termination of GDNF administration. Results TLR2 was expressed in enteric neurons, glia, and smooth muscle cells of the intestinal wall. Tlr2-/- mice had alterations in ENS architecture and neurochemical profile, intestinal dysmotility, abnormal mucosal secretion, reduced levels of GDNF in smooth muscle cells, and impaired signaling via Ret-GFRα1. ENS structural and functional anomalies were completely corrected by administration of GDNF to Tlr2-/- mice. Wild-type mice depleted of intestinal microbiota had ENS defects and GDNF deficiency, similar to Tlr2-/- mice; these defects were partially restored by administration of a TLR2 agonist. Tlr2-/- mice developed more severe colitis than wild-type mice after administration of dextran sulfate sodium or 2,4 dinitrobenzensulfonic acid; colitis was not more severe if Tlr2 -/- mice were given GDNF before dextran sulfate sodium or 2,4 dinitrobenzensulfonic acid. Conclusions In mice, TLR2 signaling regulates intestinal inflammation by controlling ENS structure and neurochemical coding, along with intestinal neuromuscular function. These findings provide information as to how defective TLR2 signaling in the ENS affects inflammatory bowel disease phenotype in humans.

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