TECHNISCHE UNIVERSITÄT MÜNCHEN Lehrstuhl für Biofunktionalität der Lebensmittel Mechanistic impact of Enterococcus faecalis metalloprotease on bacterial virulence in the context of chronic intestinal inflammation Natalie Marie Annabel Steck Vollständiger Abdruck der von der Fakultät Wissenschaftszentrum Weihenstephan für Ernährung, Landnutzung und Umwelt der Technischen Universität München zur Erlangung des akademischen Grades eines Doktors der Naturwissenschaften genehmigten Dissertation. Vorsitzender: Univ.-Prof. Dr. M. Schemann Prüfer der Dissertation: 1. Univ.-Prof. Dr. D. Haller 2. Univ.-Prof. Dr. R.F. Vogel Die Dissertation wurde am 17.11.2011 bei der Technischen Universität München eingereicht und durch die Fakultät Wissenschaftszentrum Weihenstephan für Ernährung, Landnutzung und Umwelt am 06.02.2012 angenommen. Für meine Familie und Freunde PUBLICATIONS AND PRESENTATIONS PUBLICATIONS AND PRESENTATIONS Peer-reviewed original manuscripts and reviews Steck N*, Mueller K*, Schemann M and Haller D. Bacterial proteases in IBD and IBS. Gut, 2011 (*authors equally contributed to the manuscript) Steck N , Hoffmann M, Sava IG, Kim SC, Hahne H, Tonkonogy SL, Mair K, Krueger D, Pruteanu M, Shanahan F, Vogelmann R, Schemann M, Kuster B, Sartor RB and Haller D. Enterococcus faecalis Metalloprotease Compromises Epithelial Barrier and Contributes to Intestinal Inflammation. Gastroenterology, 2011 Sep;141(3):959-71. Published abstracts Steck N , Hoffmann M, Hew Ferstl CM, Kim SC, Tonkonogy SL, Vogel RF, Sartor RB, Haller D. E. faecalis Gelatinase contributes to the development of intestinal inflammation by impairing epithelial barrier function: Role for bacteria-derived proteases in the pathology of IBD. Journal of Crohn's and Colitis Supplements - April 2010, Vol. 4, Issue 1, Pages 22-23 Steck N , Hoffmann M, Hew Ferstl CM, Kim SC, Liu B, Vogel RF, Sartor RB, Haller D. Bacterial proteases contribute to the development of chronic intestinal inflammation through the impairment of epithelial barrier functions. Gastroenterology 2009, 5(136), A21-A22 Steck N , Hoffmann M, Hew Ferstl CM, Kim SC, Tonkonogy SL, Vogel RF, Sartor RB, Haller D. Bacterial proteases contribute to the development of chronic intestinal inflammation. Int J Med Microbiol 2009; Volume: 299, p. 91 Steck N , Hoffmann M, Hew CM, Kim SC, Vogel RF, Sartor RB, Haller D. Enterococcus faecalis metalloproteinase contributes to the development of chronic intestinal inflammation through impairment of epithelial barrier integrity. Int J Med Microbiol 2008; Volume: 298, p. 96 Oral presentations Steck N , Hoffmann M, Sava I, Kim SC, Hahne H, Schemann M, Kuster B, Sartor RB, Haller D. Enterococcus faecalis Metalloprotease Compromises Epithelial Barrier and Contributes to Intestinal Inflammation. 63 rd Conference of the German Society for Hygiene and Microbiology (DGHM) e.V., 25-28.09.2011, Essen, Germany Steck N , Hoffmann M, Sava I, Kim SC, Hahne H, Mair K, Vogelmann R, Schemann M, Sartor RB, Haller D. Bacterial proteases contribute to intestinal inflammation by impairing mucosal barrier function in the susceptible host. 4th conference of German Society of Hygiene and Microbiology (DGHM) section “Microbiota, Probiota and Host”, 15.-17.04.2011, Seeon, Germany I PUBLICATIONS AND PRESENTATIONS Steck N , Hoffmann M, Hew Ferstl CM, Kim SC, Tonkonogy SL, Mair K, Vogelmann R, Schemann M, Vogel RF, Sartor BR, Haller D. Enterococcus faecalis Gelatinase impairs intestinal barrier functions: role for bacteria-derived proteases in the pathology of IBD. 7 th European Mucosal Immunology Group Meeting (EMIG), 29.09.-02.10.2010, Amsterdam, The Netherlands Steck N , Hoffmann M, Kim SC, Vogelmann R, Schemann M, Sartor BR, Haller D. E. faecalis Gelatinase contribute to the development of intestinal inflammation by impairing epithelial barrier function: role for bacteria-derived proteases in the pathology of IBD. 3 rd Joint Conference of the German Society for Hygiene and Microbiology (DGHM) e.V. and the Association for General and Applied Microbiology (VAAM), 28-31. March 2010, Hannover, Germany Steck N , Hoffmann M, Rath E, Hew Ferstl CM, Kim SC, Liu B, Vogel RF, Sartor BR, Haller D. Bacterial proteases contribute to the development of chronic intestinal inflammation through the impairment of epithelial barrier functions. 14 th International Congress of Mucosal Immunology (ICMI), 5-9. July 2009, Boston, USA Steck N , Hoffmann M, Hew Ferstl CM, Kim SC, Liu B, Vogel RF, Sartor BR, Haller D. Bacterial proteases contribute to the development of chronic intestinal inflammation through the impairment of epithelial barrier functions. Digestive Disease Week (DDW) 2009, 30.05.- 04.06.2009, Chicago, USA Steck N , Hoffmann M, Hew Ferstl CM, Kim SC, Vogel RF, Sartor BR, Haller D. Enterococcus faecalis metalloprotease contributes to the development of chronic intestinal inflammation through impairment of epithelial barrier function. 1st conference of German Society of Hygiene and Microbiology (DGHM) section “Microbiota, Probiota and Host”, 2-4.5.2008, Seeon, Germany Poster presentation Steck N , Hoffmann M, Sava I, Krüger D, Hahne H, Kim SC, Schemann M, Sartor BR, Haller D. Bacterial Gelatinase mediates loss of mucosal barrier function and contributes to intestinal inflammation. 15 th International Congress of Mucosal Immunology (ICMI), 5.-9.7.2011, Paris, France Steck N , Hoffmann M, Hew Ferstl CM, Kim SC, Tonkonogy SL, Mair K, Vogelmann R, Schemann M, Vogel RF, Sartor BR, Haller D. E. faecalis Gelatinase impairs intestinal barrier functions: role for commensal-derived proteases in the pathology of IBD. 3 rd conference of German Society of Hygiene and Microbiology (DGHM) section “Microbiota, Probiota and Host”, 18-20.06.2010, Seeon, Germany Steck N , Hoffmann M, Hew Ferstl CM, Kim SC, Tonkonogy S, Vogel RF, Sartor RB and Haller D. Bacterial proteases contribute to the development of chronic intestinal inflammation. 2nd conference of German Society of Hygiene and Microbiology (DGHM) section “Microbiota, Probiota and Host”, 23-25.04.2009, Seeon, Germany Steck N , Hoffmann M, Hew CM, Kim SC, Vogel RF, Sartor RB, Haller D. Enterococcus faecalis metalloproteinase contributes to the development of chronic intestinal inflammation through impairment of epithelial barrier integrity. 60 th conference of the German Society for Hygiene and Microbiology (DGHM) e.V., 21-24.09.2008, Dresden, Germany II PUBLICATIONS AND PRESENTATIONS Hoffmann M, Steck N , Hew CM, Kim SC, Vogel RF, Sartor RB, Haller D. Enterococcus faecalis metalloproteinase contributes to the development of chronic intestinal inflammation through impairment of epithelial barrier integrity. 45. Wissenschaftlicher Kongress der Deutschen Gesellschaft für Ernährung, 13-14.03.2008, Bonn, Germany. Grants Travel award, Organizing committee of the 7 th European Mucosal Immunology Group Meeting (EMIG), 29.09.-02.10.2010, Amsterdam, The Netherlands Awards Poster award 3rd conference of German Society of Hygiene and Microbiology (DGHM) section “Microbiota, Probiota and Host”, 18-20.06.2010, Seeon, Germany III TABLE OF CONTENTS TABLE OF CONTENTS PUBLICATIONS AND PRESENTATIONS ............................................................................. I TABLE OF CONTENTS ....................................................................................................... IV ABSTRACT ........................................................................................................................ VII ZUSAMMENFASSUNG ..................................................................................................... VIII 1 Introduction ......................................................................................................... 1 1.1 The importance of proteases .............................................................................................. 1 1.1.1 General characteristics and functions of proteases ....................................................... 1 1.1.2 Host-derived and bacterial proteases in the gut – an unexplored world of complexity . 2 1.2 Inflammatory bowel diseases ............................................................................................. 3 1.2.1 Proteases in chronic intestinal inflammation ................................................................. 4 1.2.2 Intestinal mucosal barrier dysfunction as a central key player in the pathogenesis of IBD ................................................................................................................................. 5 1.2.3 Bacteria-Host interaction: Commensal gut microbiota in IBD........................................ 7 1.2.4 Gastrointestinal infections as critical trigger for IBD ...................................................... 9 1.2.5 Bacterial proteases in IBD ........................................................................................... 10 1.2.6 Receptors for bacterial proteases in the gut ................................................................ 13 1.3 Enterococci: a narrow path between commensalism and pathogenicity .................... 15 1.3.1 Characteristics, distribution and association with disease ........................................... 16 1.3.2 Putative virulence factors in enterococci ..................................................................... 19 2 Study objective ...................................................................................................23 3 Material and Methods .........................................................................................24