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Studies on the Antimicrobial Defence of Human Hair Follicle Epithelium

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Studies on the Antimicrobial Defence of Human Hair Follicle Epithelium Aus der Klinik für Dermatologie, Venerologie und Allergologie der medizinischen Fakultät der Universität Lübeck und dem Institut für Veterinär-Anatomie des Fachbereichs Veterinärmedizin der Freien Universität Berlin Studies on the antimicrobial defence of human hair follicle epithelium Inaugural-Dissertation zur Erlangung des Grades eines Doktors der Veterinärmedizin an der Freien Universität Berlin vorgelegt von Katrin Reithmayer Tierärztin aus Landshut Berlin 2008 Journal-Nr.:3266 Gedruckt mit Genehmigung des Fachbereichs Veterinärmedizin der Freien Universität Berlin Dekan: Prof. Dr. Leo Brunnberg Erster Gutachter: Prof. Dr. Johanna Plendl Zweiter Gutachter: Prof. Dr. Ralf Paus Dritter Gutachter: Prof. Dr. Lothar H. Wieler Deskriptoren (nach CAB-Thesaurus): man, hair follicle, natural immunity, defence mechanisms, antimicrobial properties, peptides, immunohistochemistry, polymerase chain reaction Tag der Promotion: 20.04.2009 Bibliografische Information der Deutschen Nationalbibliothek Die Deutsche Nationalbibliothek verzeichnet diese Publikation in der Deutschen Nationalbibliografie; detaillierte bibliografische Daten sind im Internet über <http://dnb.ddb.de> abrufbar. ISBN: 978-3-86664-591-2 Zugl.: Berlin, Freie Univ., Diss., 2008 Dissertation, Freie Universität Berlin D 188 Dieses Werk ist urheberrechtlich geschützt. Alle Rechte, auch die der Übersetzung, des Nachdruckes und der Vervielfältigung des Buches, oder Teilen daraus, vorbehalten. Kein Teil des Werkes darf ohne schriftliche Genehmigung des Verlages in irgendeiner Form reproduziert oder unter Verwendung elektronischer Systeme verarbeitet, vervielfältigt oder verbreitet werden. Die Wiedergabe von Gebrauchsnamen, Warenbezeichnungen, usw. in diesem Werk berechtigt auch ohne besondere Kennzeichnung nicht zu der Annahme, dass solche Namen im Sinne der Warenzeichen- und Markenschutz-Gesetzgebung als frei zu betrachten wären und daher von jedermann benutzt werden dürfen. This document is protected by copyright law. No part of this document may be reproduced in any form by any means without prior written authorization of the publisher. alle Rechte vorbehalten | all rights reserved mensch und buch verlag 2009 choriner str. 85 - 10119 berlin [email protected] – www.menschundbuch.de “The pyogenic cocci, staphylococci and streptococci are constantly found on human skin, most frequently hidden in the depths of the canals of hair follicles. These micro-organisms seize every opportunity to attack the organism, producing such local lesions as acne, pimples, boils and erisipelas [SIC] or even becoming generalized in the blood and tissues as in the septicemias and pyemias. To the skin, therefore, must be assigned a very important function in the prevention of the invasion of micro-organisms, which are found constantly on the surface of the body…”(Metchnikoff 1905) Table of contents TABLE OF CONTENTS Table of contents .........................................................................................................................I Figures......................................................................................................................................IV Tables .......................................................................................................................................VI Abbreviations ..........................................................................................................................VII 1 Introduction ........................................................................................................................ 1 2 Specific questions addressed.............................................................................................. 2 3 Literature ............................................................................................................................ 3 3.1 Overview over hair follicle biology ........................................................................... 3 3.1.1 Functional anatomy of the human hair follicle .................................................. 3 3.1.2 Hair follicle cycle............................................................................................... 8 3.1.3 Hair follicle immunology................................................................................. 11 3.2 Microflora of the skin and the hair follicle .............................................................. 13 3.2.1 Resident microflora.......................................................................................... 13 3.2.2 Transient microflora......................................................................................... 14 3.2.3 Most common bacteria and fungi of skin and hair follicle infections in humans ………………………………………………………………………………...14 3.2.4 Important pathogenicity factors of bacteria ..................................................... 16 3.2.4.1 Lipopolysaccharides..................................................................................... 17 3.2.4.2 Lipoteichoic acid.......................................................................................... 18 3.2.4.3 Protein A....................................................................................................... 19 3.3 Defence mechanisms in the human skin .................................................................. 19 3.3.1 Physical barrier................................................................................................. 20 3.3.2 Chemical barrier............................................................................................... 22 3.3.3 Recognition of pathogens................................................................................. 22 3.3.4 Cellular barrier................................................................................................. 24 3.4 Antimicrobial peptides............................................................................................. 26 3.4.1 General overview............................................................................................. 26 3.4.2 Primary modes of action .................................................................................. 31 3.4.2.1 Antimicrobial activity.................................................................................. 31 3.4.2.2 Anti-endotoxin effects.................................................................................. 33 3.4.2.3 Antifungal activity........................................................................................33 I Table of contents 3.4.2.4 Antiparasitic activity.................................................................................... 33 3.4.2.5 Antiviral activity.......................................................................................... 34 3.4.3 Antimicrobial peptides as key molecules at the interface of innate and adaptive immunity… ...................................................................................................... 34 3.4.4 Additional non-immunological functions of AMPs......................................... 35 3.4.5 Antimicrobial peptides in the skin ................................................................... 36 3.5 Selected AMPs......................................................................................................... 39 3.5.1 RNase 7............................................................................................................ 39 3.5.2 Psoriasin (S100A7) .......................................................................................... 41 3.5.3 Hornerin ........................................................................................................... 43 3.5.4 LEKTI-2........................................................................................................... 44 3.6 Antimicrobial peptides in the hair follicle................................................................ 45 3.6.1 Antimicrobial peptide expression and function in the human hair follicle...... 45 3.6.2 Antimicrobial peptide expression and function in the hair follicle of other mammalian species .......................................................................................... 47 4 Material and Methods....................................................................................................... 48 4.1 Tissue specimens...................................................................................................... 48 4.2 Full thickness scalp skin organ culture..................................................................... 48 4.3 Immunohistochemistry............................................................................................. 49 4.3.1 Primary and secondary antibodies.................................................................... 50 4.3.2 Avidin-Biotin-Complex-Alkaline Phosphatase (ABC).................................... 50 4.3.3 Tyramide signal amplification (TSA) .............................................................. 51 4.3.4 Immunocytochemistry...................................................................................... 52 4.4 Microscopical equipment ......................................................................................... 52 4.5 Quantitative immunhistomorphometry .................................................................... 53 4.6 Statistical analysis.................................................................................................... 54 4.7 Isolation of outer root sheath cells ..........................................................................
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