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Jean-Paul Motta,
Luis G Bermúdez-Humarán,
Céline Deraison,
Laurence Martin,
Corinne Rolland, Perrine Rousset,
Jérôme Boue,
Gilles Dietrich,
Kevin Chapman,
Pascale Kharrat,
Jean-Pierre Vinel,
Laurent Alric,
Emmanuel Mas,
Jean-Michel Sallenave,
Philippe Langella,
Nathalie Vergnolle
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ABSTRACT: Elafin, a natural protease inhibitor expressed in healthy intestinal mucosa, has pleiotropic anti-inflammatory properties in vitro and in animal models. We found that mucosal expression of Elafin is diminished in patients with inflammatory bowel disease (IBD). This defect is associated with increased elastolytic activity (elastase-like proteolysis) in colon tissue. We engineered two food-grade strains of lactic acid bacteria (LAB) to express and deliver Elafin to the site of inflammation in the colon to assess the potential therapeutic benefits of the Elafin-expressing LAB. In mouse models of acute and chronic colitis, oral administration of Elafin-expressing LAB decreased elastolytic activity and inflammation and restored intestinal homeostasis. Furthermore, when cultures of human intestinal epithelial cells were treated with LAB secreting Elafin, the inflamed epithelium was protected from increased intestinal permeability and from the release of cytokines and chemokines, both of which are characteristic of intestinal dysfunction associated with IBD. Together, these results suggest that oral delivery of LAB secreting Elafin may be useful for treating IBD in humans.
Science translational medicine 10/2012; 4(158):158ra144. · 7.80 Impact Factor
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Jean-Paul Motta,
Laurent Magne,
Delphyne Descamps,
Corinne Rolland,
Camila Squarzoni-Dale, Perrine Rousset,
Laurence Martin,
Nicolas Cenac,
Viviane Balloy,
Michel Huerre,
Leopold F Fröhlich,
Dieter Jenne,
Julien Wartelle,
Azzaq Belaaouaj,
Emmanuel Mas,
Jean-Pierre Vinel,
Laurent Alric,
Michel Chignard,
Nathalie Vergnolle,
Jean-Michel Sallenave
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ABSTRACT: Colonic tissues of patients with inflammatory bowel disease have been reported to have increased proteolytic activity, but no studies have clearly addressed the role of the balance between proteases and antiproteases in the pathogenesis of colitis. We investigated the role of Elafin, a serine protease inhibitor expressed by skin and mucosal surfaces in human inflammatory conditions, and the proteases neutrophil elastase (NE) and proteinase-3 (PR-3) in mice with colitis.
We studied mice with heterozygous disruptions in NE and PR-3, mice that express human elafin (an inhibitor of NE and PR-3), and naïve mice that received intracolonic adenoviral vectors that express elafin. Trinitrobenzene sulfonic acid (TNBS) or dextran sodium sulphate (DSS) was used to induce colitis. Protease, cytokine levels, and NF-κB activity were measured in colons of mice. Caco-2 and HT29 cells were studied in assays for cytokine expression, permeability, and NF-κB activity.
Elafin expression or delivery re-equilibrated the proteolytic balance in inflamed colons of mice. In mice given TNBS or DSS, transgenic expression of elafin or disruption of NE and PR-3 protected against the development of colitis. Similarly, adenoviral delivery of Elafin significantly inhibited inflammatory parameters. Elafin modulated a variety of inflammatory mediators in vitro and in vivo and strengthened intestinal epithelial barrier functions.
The protease inhibitor Elafin prevents intestinal inflammation in mouse models of colitis and might be developed as a therapeutic agent for inflammatory bowel disease.
Gastroenterology 01/2011; 140(4):1272-82. · 11.68 Impact Factor
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ABSTRACT: Ligand-gated calcium channels have been reported to be involved in the pathogenesis of inflammatory bowel disease. One family member, transient receptor potential vanilloid 4 (TRPV4), is activated by arachidonic acid derivatives that might be released on inflammation, yet its role in gastrointestinal inflammation has not been characterized. We investigated whether TRPV4 activation participates in intestinal inflammation and its expression and functions in the gastrointestinal tract.
TRPV4 expression was studied in human colon samples, human intestinal epithelial cell lines (Caco-2 and T84), and inflamed colons of mice. Calcium mobilization and cytokine release were analyzed in intestinal epithelial cells exposed to the selective TRPV4 agonist 4α-phorbol-12,13-didecanoate (4αPDD). Mice were killed 3, 6, or 24 hours after intracolonic administration of 4αPDD; inflammatory parameters were measured in their colon tissues, and paracellular colonic permeability was measured by the passage of (51)Cr-EDTA from the colon lumen to the blood.
High levels of TRPV4 were detected in Caco-2 cells and in epithelial cells of human colon tissue samples; its expression was up-regulated in colons from inflamed mice compared with noninflamed control mice. Administration of 4αPDD to Caco-2 and T84 cells caused a dose-dependent increase in intracellular calcium concentration and chemokine release. In mice, intracolonic administration of 4αPDD caused colitis to develop 3 to 6 hours later; inflammation resolved by 24 hours. Increased colonic permeability was observed in vivo 3 hours after intracolonic administration of 4αPDD.
TRPV4 is expressed and functional in intestinal epithelial cells; its activation in the gastrointestinal tract causes increases in intracellular calcium concentrations, chemokine release, and colitis.
Gastroenterology 09/2010; 140(1):275-85. · 11.68 Impact Factor
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ABSTRACT: Given the anti-inflammatory effects of insulin in human and animal studies done in vivo and given the signaling pathways in common between insulin and the protease-activated receptor 2 (PAR(2)), a G protein-coupled receptor, we hypothesized that insulin would have an impact on the inflammatory actions of PAR(2). We found that low doses or concentrations of insulin in the subnanomolar range reduced PAR(2)-induced inflammation in a murine paw edema model, attenuated PAR(2)-induced leukocyte trafficking in mouse intestinal venules, and reduced PAR(2) calcium signaling in cultured dorsal root ganglion neurons and endothelial cells. This effect of insulin to attenuate PAR(2)-mediated inflammation was reversed when cells were preincubated with LY294002 (a PI3K inhibitor) and GF 109203X (a pan-protein kinase C inhibitor). The enhanced inflammatory effect of PAR(2) observed in vivo in an insulin-deficient murine type 1 diabetes model was attenuated by the local administration of insulin at the inflammatory site. Our data point to an anti-inflammatory action of insulin that targets the acute innate inflammatory response triggered by PAR(2).
The Journal of Immunology 03/2010; 184(5):2702-9. · 5.79 Impact Factor
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ABSTRACT: Protease-activated receptor-2 (PAR(2)), a receptor highly expressed in the respiratory tract, can influence inflammation at mucosal surfaces. Although the effects of PAR(2) in the innate immune response to bacterial infection have been documented, knowledge of its role in the context of viral infection is lacking. We thus investigated the role of PAR(2) in influenza pathogenesis in vitro and in vivo. In vitro, stimulation of PAR(2) on epithelial cells inhibited influenza virus type A (IAV) replication through the production of IFN-gamma. In vivo, stimulation of PAR(2) using specific agonists protected mice from IAV-induced acute lung injury and death. This effect correlated with an increased clearance of IAV in the lungs associated with increased IFN- gamma production and a decreased presence of neutrophils and RANTES release in bronchoalveolar fluids. More importantly, the protective effect of the PAR(2) agonist was totally abrogated in IFN- gamma-deficient mice. Finally, compared with wild-type mice, PAR(2)-deficient mice were more susceptible to IAV infection and displayed more severe lung inflammation. In these mice higher neutrophil counts and increased RANTES concentration but decreased IFN- gamma levels were observed in the bronchoalveolar lavages. Collectively, these results showed that PAR(2) plays a protective role during IAV infection through IFN-gamma production and decreased excessive recruitment of inflammatory cells to lung alveoli.
The Journal of Immunology 07/2009; 182(12):7795-802. · 5.79 Impact Factor
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Jean-Paul Motta,
Laurent Magne,
Delphyne Descamps,
Corinne Rolland,
Camila Squarzoni Dale, Perrine Rousset,
Viviane Balloy,
Michel Huerre,
Dieter Jenne,
Julien Wartelle,
Azzaq Belaaouaj,
Emmanuel Mas,
Jean-Pierre Vinel,
Laurent Alric,
Michel Chignard,
Nathalie Vergnolle,
Jean-Michel Sallenave
[show abstract]
[hide abstract]
ABSTRACT: BACKGROUND & AIMS:: Colon tissues of patients with inflammatory bowel disease (IBD) have been reported to have increased proteolytic activity, but no studies have clearly addressed the protease to anti-protease balance in the pathogenesis of colitis. We investigated the role of Elafin, a serine protease inhibitor expressed by skin and mucosal surfaces in human inflammatory conditions, and the proteases neutrophil elastase (NE) and proteinase-3 (PR-3), in mice with colitis. METHODS:: We studied mice with heterozygous disruptions in NE and PR-3and 2 strains of mice that express transgenic human elafin (an inhibitor of NE and PR-3). Trinitrobenzene sulfonic acid (TNBS) or dextran sodium sulphate (DSS) was used to induce colitis. Protease and cytokine levels were measured in colonic tissues collected from the mice. CaCO(2) and HT29 cells were studied in assays for cytokine expression and permeability. RESULTS:: Mice that expressed transgenic elafin developed less colitis and had reduced levels of proteases than wild-type mice following administration of TNBS or DSS. Expression of elafin did not modify activities of elastase or PR-3, but inhibited their increase upon the induction of colitis. Mice that expressed reduced levels of NE and PR-3 were protected against DSS-induced colitis. Elafin expression altered the patterns of inflammatory mediators and strengthened intestinal epithelial barrier functions in cells and colonic tissues from mice. CONCLUSIONS:: The protease inhibitor Elafin prevents intestinal inflammation in a mouse model of colitis and might be developed as a therapeutic agent for IBD.
Gastroenterology.
