E Quévrain1,2,3, M A Maubert1,2,3,4, C Michon5,6, H Sokol1,2,3,5,7, P Seksik1,2,3,7, F Chain5,6, R Marquant1,3,8, J Tailhades1,3,8, S Miquel5,6, L Carlier1,3,8, L G Bermúdez-Humarán5,6, B Pigneur1,2,3, O Lequin1,3,8, P Kharrat5,6, G Thomas1,2,3, D Rainteau1,2,3,4, C Aubry5,6, N Breyner5,6, C Afonso9, S Lavielle1,3,8, J-P Grill1,2,3, G Chassaing1,3,8, J M Chatel5,6, G Trugnan1,2,3,4, R Xavier10, P Langella5,6. 1. Sorbonne Universités, UPMC Univ Paris 06, LBM, 27 rue de Chaligny, F-75012, Paris, France. 2. INSERM-ERL 1157 and Inflammation-Immunopathology-Biotherapy Department (DHU i2B), CHU Saint-Antoine 27 rue de Chaligny, F-75012 Paris, France. 3. CNRS, UMR 7203 LBM, F-75005, Paris, France. 4. APHP, Hôpital Saint Antoine - Département PM2 Plateforme de Métabolomique, Peptidomique et dosage de Médicaments, F-75012 Paris, France. 5. INRA, UMR1319 Micalis, F-78350 Jouy-en-Josas, France. 6. AgroParisTech, UMR Micalis, F-78350 Jouy-en-Josas, France. 7. APHP, Hôpital Saint Antoine - Service de Gastroentérologie et nutrition, F-75012 Paris, France. 8. Ecole Normale Supérieure- PSL Research University, Département de Chimie 24 rue Lhomond, F-75005 Paris, France. 9. Université de Rouen, UMR 6014 COBRA / IRCOF, F-76130 Mont Saint Aignan, France. 10. Center for Systems Biology, Massachusetts General Hospital, Boston, Massachusetts, USA.
Abstract
BACKGROUND: Crohn's disease (CD)-associated dysbiosis is characterised by a loss of Faecalibacterium prausnitzii, whose culture supernatant exerts an anti-inflammatory effect both in vitro and in vivo. However, the chemical nature of the anti-inflammatory compounds has not yet been determined. METHODS: Peptidomic analysis using mass spectrometry was applied to F. prausnitzii supernatant. Anti-inflammatory effects of identified peptides were tested in vitro directly on intestinal epithelial cell lines and on cell lines transfected with a plasmid construction coding for the candidate protein encompassing these peptides. In vivo, the cDNA of the candidate protein was delivered to the gut by recombinant lactic acid bacteria to prevent dinitrobenzene sulfonic acid (DNBS)-colitis in mice. RESULTS: The seven peptides, identified in the F. prausnitzii culture supernatants, derived from a single microbial anti-inflammatory molecule (MAM), a protein of 15 kDa, and comprising 53% of non-polar residues. This last feature prevented the direct characterisation of the putative anti-inflammatory activity of MAM-derived peptides. Transfection of MAM cDNA in epithelial cells led to a significant decrease in the activation of the nuclear factor (NF)-κB pathway with a dose-dependent effect. Finally, the use of a food-grade bacterium, Lactococcus lactis, delivering a plasmid encoding MAM was able to alleviate DNBS-induced colitis in mice. CONCLUSIONS: A 15 kDa protein with anti-inflammatory properties is produced by F. prausnitzii, a commensal bacterium involved in CD pathogenesis. This protein is able to inhibit the NF-κB pathway in intestinal epithelial cells and to prevent colitis in an animal model. Published by the BMJ Publishing Group Limited. For permission to use (where not already granted under a licence) please go to http://www.bmj.com/company/products-services/rights-and-licensing/
BACKGROUND:Crohn's disease (CD)-associated dysbiosis is characterised by a loss of Faecalibacterium prausnitzii, whose culture supernatant exerts an anti-inflammatory effect both in vitro and in vivo. However, the chemical nature of the anti-inflammatory compounds has not yet been determined. METHODS: Peptidomic analysis using mass spectrometry was applied to F. prausnitzii supernatant. Anti-inflammatory effects of identified peptides were tested in vitro directly on intestinal epithelial cell lines and on cell lines transfected with a plasmid construction coding for the candidate protein encompassing these peptides. In vivo, the cDNA of the candidate protein was delivered to the gut by recombinant lactic acid bacteria to prevent dinitrobenzene sulfonic acid (DNBS)-colitis in mice. RESULTS: The seven peptides, identified in the F. prausnitzii culture supernatants, derived from a single microbial anti-inflammatory molecule (MAM), a protein of 15 kDa, and comprising 53% of non-polar residues. This last feature prevented the direct characterisation of the putative anti-inflammatory activity of MAM-derived peptides. Transfection of MAM cDNA in epithelial cells led to a significant decrease in the activation of the nuclear factor (NF)-κB pathway with a dose-dependent effect. Finally, the use of a food-grade bacterium, Lactococcus lactis, delivering a plasmid encoding MAM was able to alleviate DNBS-induced colitis in mice. CONCLUSIONS: A 15 kDa protein with anti-inflammatory properties is produced by F. prausnitzii, a commensal bacterium involved in CD pathogenesis. This protein is able to inhibit the NF-κB pathway in intestinal epithelial cells and to prevent colitis in an animal model. Published by the BMJ Publishing Group Limited. For permission to use (where not already granted under a licence) please go to http://www.bmj.com/company/products-services/rights-and-licensing/
Authors: L A Dieleman; M J Palmen; H Akol; E Bloemena; A S Peña; S G Meuwissen; E P Van Rees Journal: Clin Exp Immunol Date: 1998-12 Impact factor: 4.330
Authors: Denise Kelly; Jamie I Campbell; Timothy P King; George Grant; Emmelie A Jansson; Alistair G P Coutts; Sven Pettersson; Shaun Conway Journal: Nat Immunol Date: 2003-12-21 Impact factor: 25.606