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Old Herborn University Monograph 16 Old Herborn University Seminar Monograph 16. HOST MICROFLORA CROSSTALK EDITORS: PETER J. HEIDT TORE MIDTVEDT VOLKER RUSCH DIRK VAN DER WAAIJ Old Herborn University Seminar Monograph 16 ISBN 3-923022-27-1 ISSN 1431-6579 COPYRIGHT © 2003 BY HERBORN LITTERAE ALL RIGHTS RESERVED NO PART OF THIS PUBLICATION MAY BE REPRODUCED OR TRANSMITTED IN ANY FORM OR BY ANY MEANS, ELECTRONIC OR MECHANICAL, INCLUDING PHOTOCOPY, RECORDING, OR ANY INFORMATION STORAGE AND RETRIEVAL SYSTEM, WITHOUT PERMISSION IN WRITING FROM THE PUBLISHER EDITORS: Peter J. Heidt, Ph.D., B.M. Department of Animal Science Biomedical Primate Research Centre (BPRC) Lange Kleiweg 139 2288 GJ - Rijswijk The Netherlands Tore Midtvedt, M.D., Ph.D. Department of Medical Microbial Ecology Karolinska Insttute von Eulers väg 5 S 171 77 Stockholm Sweden Volker Rusch, Dr. rer. nat. Institute for Integrative Biology Kornmarkt 2 D-35745 Herborn-Dill Germany Dirk van der Waaij, M.D., Ph.D. Professor emeritus, University of Groningen Hoge Hereweg 50 9756 TJ - Glimmen The Netherlands Verlag wissenschaftlicher Schriften und Bücher Am Kornmarkt 2 Postfach 1664 D-35745 Herborn-Dill Germany Telephone: +49 - 2772 - 921100 Telefax: +49 - 2772 - 921101 Contents ——————————————————————————————————————— Participating authors V I. THE GUT IMMUNE SYSTEM AND THE MUCOSAL BACTERIA (Agnes E. Wold) 1 Summary ……………………………………………………………….. 1 IgA ……………………………………………………………………… 1 T cells…………………………………………………………………... 2 Induction of mucosal immune responses……………………………….. 2 Importance of gut flora on the specific immune system ………………… 3 The transient nature of the response to gut bacteria …………………….. 4 Immune response to food proteins ……………………………………… 5 Oral tolerance …………………………………………………………… 5 Mechanisms for oral tolerance ………………………………………….. 6 The normal microflora and oral tolerance……………………………….. 7 Influence of the commensal flora on innate immunity…………………... 7 Different bacteria elicit different innate responses ……………………… 8 Are we intolerant because of a faulty intestinal microflora? …………….. 9 Literature ……………………………………………………………….. 10 II. DEFENSINS AND DEFENSIN-LIKE MOLECULES: ANTIBACTERIAL MODE OF ACTION (Hilde Ulvatne, Ørjan Samuelsen, and Lars Harry Vorland) 17 Summary ……………………………………………………………….. 17 Introduction …………………………………………………………….. 17 Discussion………………………………………………..…………….. 21 Conclusion ……………………………………………….…………….. 25 Literature ……………………………………………………………….. 26 III. DEFENSINS AND BACTERIA, A QUESTION OF "LIVE OR LET DIE"? (Lars-Göran Axelsson) 33 Summary ……………………………………………………………….. 33 Introduction …………………………………………………………….. 33 Innate immunity of the gastrointestinal tract ……………………………. 34 Antimicrobial peptides, molecular properties and mode of action ………. 35 Small intestinal microbial peptides ……………………………………… 36 Pharmaceutical applications …………………………………………….. 38 Literature ……………………………………………………………….. 38 I Contents (continued) ——————————————————————————————————————— IV. PHENOTYPIC EXPRESSIONS IN THE SMALL INTESTINE (Elisabeth Norin)4 3 Introduction …………………………………………………………….. 43 Cell kinetics …………………………………………………………….. 44 Intestinal motility ……………………………………………………….. 45 Short chain fatty acids ………………………………………………….. 45 Intestinal tryptic activity ………………………………………………… 46 Bilirubin and urobilins………………………………………………….. 47 Concluding remarks…………………………………………………….. 47 Literature ……………………………………………………………….. 47 V. MICROBIAL P450: DOES IT EXIST, AND WHAT CAN IT MEAN? (Tore Midtvedt) 51 Summary ……………………………………………………………….. 51 Nomenclature …………………………………………………………… 51 P450s and metabolism of xenobiotics ……………………..…………... 51 P450s and evolution ……………………………………………………. 52 Xenobiotic metabolism and substrate specificity ……………………….. 53 Localisation of P450s…………………………………………………… 54 P450s and microorganisms ……………………………………………. 54 P450s and specific members of the intestinal flora …………………….. 54 Literature ……………………………………………………………….. 55 VI. A LINK BETWEEN MUCOSAL REGULATORY LYMPHOCYTES AND CHILDHOOD FOOD ALLERGY (Simon H. Murch) 57 Introduction …………………………………………………………….. 57 The increase in the incidence of food allergies………………………….. 57 Does IgE or IgA determine sensitisation? ………………………………. 57 Genetic predisposition to food allergy ………………………………….. 58 Regulation of IgE responses ……………………………………………. 58 Multiple food intolerance, food-allergic dysmotility and the eosinophil response ………………………………………………………………… 59 Demographics of allergic sensitisation: The role of enteric challenges….. 59 Mucosal challenges in children born in developing countries…………… 60 Contrasting changes in early-life gut flora in the developed-world child . 61 The development of enteric tolerance …………………………………… 62 Mechanisms of oral tolerance: A central role for NF-κB ……………….. 63 Is food allergy regulated to defective generation of regulatory lymphocytes? …………………………………………………………… 64 Literature ……………………………………………………………….. 65 II Contents (continued) ——————————————————————————————————————— VII. SYNBIOTIC TREATMENT IN CLINICAL PRAXIS (Stig Bengmark)6 9 Introduction …………………………………………………………….. 69 Fermented food has outstanding quality………………………………… 69 Flora boosts the immune system of the host ……………………………. 71 Prebiotic fibres are essential ……………………………………………. 72 Probiotics in diarrhoea in children ……………………………………… 73 Probiotics - and prebiotics - in inflammatory bowel disease (IBD)……... 74 Probiotics in Helicobacter pylori infections …………………………….. 75 Synbiotics in ICU patients ……………………………………………… 76 Flora important on all body surfaces……………………………………. 77 Gut ecology and health - future aspects…………………………………. 78 Literature………………………………………………………………… 79 VIII. DEVELOPING AN IN VITRO MODEL ON THE INVESTIGATION OF THE CROSSTALK AMONG BACTERIA, ENTEROCYTES AND LEUKOCYTES NEAR THE INTESTINAL MUCOSA (Alexandr Parlesak and Christiane Bode) 83 Summary …………………………………………………………..…... 83 Introduction …………………………………………………………….. 83 Material and methods …………………………………………………… 87 First results and conclusion …………………………………………….. 87 Literature ……………………………………………………………….. 89 IX. HOW MUCOSAL IMMUNITY IS CONTROLLED BY LOCAL FACTORS (Stefan C. Meuer) 91 The author's literature concerning this field…………………………….. 93 X. OLD HERBORN UNIVERSITY SEMINAR ON HOST MICROFLORA CROSSTALK: MINUTES AND OVERVIEW OF THE DISCUSSIONS (Dirk van der Waaij)9 5 Elaine Vaughan: Approaches to investigate the diversity and functionality of intestinal microbes……………………………………... 95 Vanya E. Grant: Flow cytometry - can it help to analyse complex biosystems? …………………………………………………………….. 96 Hilde Ulvatne: Defensins and defensin-like molecules: Antibacterial mode of action ………………………………………………………….. 96 Lars-Göran Axelsson: Defensins and bacteria, a question of "live or let die?"…………………………………………………………. 98 Agnes Wold: Mucosal immunology ……………………………………. 99 III Contents (continued) ——————————————————————————————————————— Simon Murch: A link between mucosal regulatory lymphocytes and childhood food allergy ………………………………………………… 100 Stig Bengmark: Synbiotic treatment in clinical praxis…………………. 100 Elisabeth Norin: Phenotypic expressions in the small intestine ……….. 100 Barbara H. Iglewski: Quorum sensing in Pseudomonas aeruginosa …. 101 Alexander Parlesak: Developing an in vitro model on the investigation of the crosstalk among bacteria, enterocytes and leukocytes near the intestinal mucosa ……………………………………………………… 101 IV Participating authors ——————————————————————————————————————— Lars-Göran Axelsson, Ph.D., Microbiology and Tumorbiology Center, Gas­ trobiology Unit, Division of Molecular Pathology, Karolinska Institute, Box 280, Theorells väg 3, S-171 77 Stockholm, Sweden. Stig Bengmark, M.D., Ph.D., Lund University, Ideon Research Park, Schelevägen 18, S-22 370 Lund, Sweden. Stefan C. Meuer, M.D., Ph.D., Institute for Immunology, Ruprecht-Karls- University, Im Neuenheimer Feld 305, D-69210 Heidelberg, Germany. Tore Midtvedt, M.D., Ph.D., Department of Medical Microbial Ecology, Karolinska Institute, von Eulers väg 5, Box 60 400, S-17177 Stockholm, Sweden. Simon H. Murch, B.Sc. Ph.D., Royal Free and University College School of Medicine, Royal Free Campus, Rowland Hill Street, London NW3 2PF, UK. Elisabeth Norin, Ph.D., Microbiology and Tumorbiology Center, Karolinska Institute, von Eulers väg 5, S-171 77 Stockholm, Sweden. Alexandr Parlesak, Ph.D., Institute for Biological Chemistry and Nutrition, Department Physiology of Nutrition, Hohenheim University, Garbenstraße 28, D-70599 Stuttgart, Germany. Hilde Ulvatne Marthinsen, M.D., Ph.D., Department of Medical Micro­ biology, University Hospital of North Norway, N-9038 Tromsø, Norway. Dirk van der Waaij, M.D., Ph.D., Hoge Hereweg 50, 9756 TJ Glimmen, The Netherlands. Agnes E. Wold, M.D., Ph.D., Department of Clinical Bacteriology, Göteborg University, Guldhedsgatan 10a, S-413 46 Göteborg, Sweden. V VI THE GUT IMMUNE SYSTEM AND THE MUCOSAL BACTERIA AGNES E. WOLD Department of Clinical Bacteriology, Göteborg University, Göteborg, Sweden. SUMMARY The gut associated immune system harbours the vast majority of all lymphoid cells in the human body. This corresponds to the fact that most antigens reach us via the gut. The mucosa of the upper airways and the gastro-intestinal tracts also host a rich normal bacterial flora, which serves as a stimulus to the immune system. There are many indi­ cations that the normal intestinal microflora affects the way in which other antigens, e.g. food antigens and other innocuous antigens, are handled. Thus, alterations in intestinal colonisation pattern might have predisposed for allergies and other hypersensitivity reactions. In this review, the immune system of the gut and its relation to the
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