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wissenschaft braucht emotionen
FORSCHUNGSBERICHT 2013/2014 WISSENSCHAFT BRAUCHT EMOTIONEN WISSENSCHAFTLICHER BEIRAT/SCIENTIFIC ADVISORY BOARD Prof. Dr. Claus Vogelmeier (Vorsitzender) Klinik für Pneumologie Universitätsklinikum Gießen-Marburg, Marburg Prof. Dr. Axel Brakhage (stv. Vorsitzender) Leibniz-Institut für Naturstoff-Forschung u. Infektionsbiologie (HKI), Jena Prof. Dr. Christoph Dehio Infection Biology Universität Basel, Basel Prof. Dr. Bernhard Fleischer Bernhard-Nocht-Institut für Tropenmedizin, Hamburg Prof. Dr. Inken König Institut für Medizinische Biometrie und Statistik Universität zu Lübeck, Lübeck Prof. Dr. Gisa Tiegs Institut für Experimentelle Immunologie und Hepatologie Universitätsklinikum Eppendorf, Hamburg Prof. Dr. Erika von Mutius Dr. von Haunersches Kinderspital der Ludwig-Maximilians-Universität München, München IMPRESSUM Herausgeber Prof. Dr. S. Ehlers Koordination Dr. B. C. Brand Bilder Fotolia S. 5 @ Sergey Nivens S. 6 @ Westend S. 8 @ Victoria S. 16 @ Sagittaria S. 76 @ 2867025 S. 77 @ 64660 FZB S. 13; S. 15; S.17; S. 16; S. 19 (H. Haas); S. 25 Design STILPUNKT3 Designbüro Hamburg Druck Partner Werbung & Druck Hamburg Seite 5 /Inhalt VORWORT Seite 6 MAGAZIN Leibniz macht es Spaß im Job Farewell Highlights Menschen • People möglich: Fächerüber- Zufriedene Mitarbeiter Sabine Rüsch-Gerdes Presse 2013 greifend gegen Infekti- sind essentiell für Jabbar Ahmed 2013 2014 onskrankheiten den Erfolg Martin Ernst 2014 Helmut Haas Seite 11-12 Seite 8-10 Seite 13-15 Seite 20-23 Seite 16-19 Seite 24-27 RESEARCH REPORTS MEDICINE PRA ASTHMA AND ALLERGY PRA INFECTIONS 52 Center for Clinical Studies 68 Bioanalytical Chemistry 28 Biochemical Immunology Bioinformatics 30 Clinical and Molecular Allergology 54 Clin. and Exp. Pathology 70 Biophysics32 Experimental Pneumology 56 National Reference Center Cellular Microbiology 34 Innate Immunity 58 for Mycobacteria 72 Cellular Pneumology 36 Invertebrate Models 60 Clinical Infectious Diseases 38 Mouse Models of Asthma 62 Coinfection 40 Mucosa Immunology & Diagnostics64 CENTRAL UNITS Immunobiophysics 42 Structural Biochemistry 66 Fluorescence Cytometry 74 Infection Immunology 44 Microbial Interface Biology 46 Models of Inflammation 48 Molecular Mycobacteriology 50 FACTS & FIGURES Funding Seite 76 Patents and Licenses Seite 76 Academic Degrees and Professional Qualifications Seite 76 Conferences and Workshops Seite 77 Peer Review Publications Seite 77 Books and Book Articles Seite 77 National Networks Seite 77 International Networks Seite 77 Organization Chart Seite 78 IMPRESSUM WISSENSCHAFT BRAUCHT EMOTIONEN ––––––––––––––––––– SCIENCE NEEDS EMOTIONS PROF. LUDWIG SCHULTZ, GDNÄ Seite 7 / Vorwort • Preface SEHR GEEHRTE LESERIN, SEHR GEEHRTER LESER ––––––––––––––––––– DEAR READER D T Der Magazinteil berichtet über Aktivitäten des Zentrums, ein developing structured and versatile well-being-programs for strukturiertes und vielseitiges Well-Being-Programm für die employees, events affecting the Borstel community, important Beschäftigten aufzubauen, bewegende Ereignisse und wich- research results, scientific networks and new research teams. tige Forschungsergebnisse, Verbundaktivitäten und neue By way of examples, hand-picked projects of the research Forschungsgruppen. Exemplarisch ausgewählte Projekte der groups and clinical units illustrate both the core competences Forschungsteams zeigen sowohl die Kernkompetenzen auf als and the achieved accomplishments. er vorliegende Forschungsbericht informiert in kompakter Form über die wissenschaftlichen Aktivitäten des Forschungszentrums Borstel in den Jahren 2013 und 2014. auch die erbrachten Leistungen. he present report informs about the scientific activities of the Research Center Borstel in the years 2013 and 2014 in a compact form. The magazine section reports on activities In this way, we would like to give an understanding of our Auf diese Weise möchten wir nicht nur versierten Kolleginnen research approach and objectives not only to experienced col- und Kollegen, sondern auch einer breiteren Öffentlichkeit unse- leagues but also to a broader public. In addition, this report re wissenschaftlichen Fragestellungen und Ziele näher bringen, shows the progress we have made during the last two years in und darüber hinaus auch die Fortschritte aufzeigen, die wir in contributing to solving medical problems relevant to society. den beiden vergangenen Jahren bei der Lösung gesellschaftlich The Research Center Borstel is the Lung Center of the Leibniz relevanter medizinischer Probleme erzielt haben. Association. In this context, “Leibniz” stands for application- Das FZB ist das Lungenforschungszentrum der LeibnizGemeinschaft. „Leibniz“ steht in diesem Zusammenhang für and patient-oriented basic biomedical research; lung research focuses on “infections” and “asthma and allergy”. anwendungs- und patientenorientierte Grundlagenforschung The Borstel holistic approach spanning from molecular struc- („theoria cum praxi“); die Lungenforschung konzentriert sich tures via cellular processes to the application of complex model auf die Schwerpunkte „Infektionen“ und „Asthma und Aller- systems, opens up an entirely new understanding of causal re- gie“. Die Borstel-typische ganzheitliche Betrachtungsweise, lationships. Some of our main goals in pulmonary research are: die von molekularen Strukturen bis hin zum komplexen biologischen System des Menschen selbst reicht, ermöglicht ein •Identification of new target structures for anti-bacterial, an- völlig neues Verständnis für Zusammenhänge. Dabei streben ti-inflammatory and anti-allergic preventive and therapeutic wir u.a. die folgenden Ziele an: interventions. •Exploiting our knowledge of mechanisms of resistance, vir- 1.Identifizierung neuer Zielstrukturen für anti-bakterielle, antientzündliche und anti-allergische Prophylaxen und Therapien. 2.Nutzung der Kenntnis von Resistenz-, Virulenz-, Persistenz- und Reaktivierungsmechanismen für Therapien und Prognose. 3.Identifikation von Immunmodulatoren zur Prophylaxe und ulence, persistence and reactivation for improving therapy and prognosis. •Identification of immune modulators for prophylaxis and therapy of infectious and non-infectious, acute and chronic inflammatory conditions. Therapie von infektiösen und nicht-infektiösen Entzündungen und deren chronischer Ausprägung. Change is positive – and excellent research will ultimately benefit society! We are proud of the excellent work of all of our Veränderung ist unser Motto – für eine exzellente Forschung colleagues, and we are deeply grateful to all friends and sup- zum Nutzen der Gesellschaft! Stolz sind wir auf die hervorra- porters from science, politics, and industry. With your support gende Arbeit aller unserer Mitarbeiterinnen und Mitarbeiter, we are looking optimistically into the future! ein herzlicher Dank gilt allen Freunden und Förderern aus Wissenschaft, Politik und Wirtschaft. Mit Ihrer Unterstützung blicken wir optimistisch in die Zukunft! Stefan Ehlers, Heinz Fehrenbach, Stefan Niemann, Stefan Ehlers, Heinz Fehrenbach, Stefan Niemann, Frank Petersen, Ulrich Schaible, Peter Zabel Frank Petersen, Ulrich Schaible, Peter Zabel –––––––––––––––– Autor: Stefan Ehlers und Ulrich Schaible Bild: ©Victoria, fotolia –––––––––––––––– LEIBNIZ MACHT ES MÖGLICH: FÄC HERÜBERGREIFEND GEGEN INFEKTIONSKRANKHEITEN ––––––––––––––––––– Der Leibniz-Forschungsverbund Infections ’21 (Infektionen im 21. Jahrhundert) nimmt Fahrt auf. Seite 9 / Reportage D ie Leibniz-Gemeinschaft fördert im Rahmen der Förder- III. die Übertragung von Erregern wie z. B. Grippeviren über linie Strategische Vernetzung ab 2015 für 4 Jahre ein Wasservögel und möglicherweise kontaminierte Oberflä- vom Forschungszentrum Borstel initiiertes und koordiniertes chengewässer (WATER); Verbundvorhaben mit insgesamt 600.000 Euro. Der Leibniz IV. die klimabedingte Ausbreitung von Insekten und Zecken Forschungsverbund mit dem Titel INFECTIONS’21 – Transmis- und durch sie übertragene Krankheitserreger, die bisher nicht sion Control of Infections in the 21st Century hat es sich zur bei uns vorkommen (VECTORS). Aufgabe gemacht Kontrolle, Prävention und Bekämpfung von Infektionskrankheiten weit über den biomedizinischen Rah- Während im MANtoMAN-Projekt Sozialwissenschaftler und men hinaus fächerübergreifend anzugehen. Für dieses inter- Biomediziner gemeinsam Methoden entwickeln, um schwierig disziplinäre Vorhaben hat das FZB 13 weitere Leibniz-Institute zugängliche Randgruppen zu erreichen und die Ausbreitungs- der Sektionen Lebens-, Umwelt und Sozialwissenschaften so- dynamik wichtiger Infektionen zu erfassen, studieren bei AIR wie 3 externe Partner um sich geschart. Klimaforscher, Physiker und Infektionsbiologen um klimati- Dass Gesundheit mehr ist als nur körperliches Wohlbefin- sche und physikalische Bedingungen für die Verbreitung von den, ist jedem spätestens seit der WHO-Definition von Gesund- Erregern über die Luft zu definieren und retrospektiv einen heit bekannt. Ausbruch der Legionärskrankheit, die ihren Ursprung in einer ––––––––––––––––––– Health is a state of complete physical, mental and social well-being and not merely the absence of disease or infirmity. Preamble to the Constitution of the World Health Organization as adopted by the International Health Conference, New York, 19-22 June, 1946; signed on 22 July 1946 by the representatives of 61 States (Official Records of the World Health Organization, no. 2, p. 100) and entered into force on 7 April 1948. ––––––––––––––––––– Die Entwicklungen der Ebola-Epidemie in Westafrika und Kühlanlage einer Brauerei hatte, aufzuarbeiten. Die an WATER beteiligten Biomediziner, Umwelt- und Naturwissenschaftler untersuchen Wasservögel und die von ihnen frequentierten Gewässer, die als Hot Spots für den Austausch von Infektionserregern zwischen verschiedenen Tierarten dienen können. VECTORS integriert Umwelt-, Klima-, Agrar- und biomedizinische Forschung aber auch die Öffentlichkeit, um die Ausbreitung neuer Krankheitserreger, die von Arthropoden übertragen werden, zu verfolgen und entsprechende Frühwarnsysteme zu etablieren, um ihre Ankunft in Deutschland nachzuweisen. ihre sozialen und wirtschaftlichen Folgeerscheinungen macht es offenkundig: Neben der Biomedizin sind weitere Disziplinen Die vier ausgewählten Themen werden anhand definierter gefordert, um den umfassenden Herausforderungen durch In- Projekte innerhalb sogenannter Interdisciplinary Research fektionskrankheiten zu begegnen. Unwissenheit über die An- Groups (IRG) beforscht. Dazu werden interdisziplinäre Dok- steckungswege fördert die Verbreitung der Infektion, Angst vor torarbeiten vergeben, die von Wissenschaftlerinnen und Wis- sozialer Isolierung führt zur Vermeidung von Schutzmaßnah- senschaftlern aus den unterschiedlichen Leibniz-Instituten men, der Mangel an Behandlungszentren verdrängt die medi- gemeinsam betreut werden. Jedes der Projekte wird von den zinische Primärversorgung, Angst vor Ansteckung beschränkt Promovierenden aber auch ihren Betreuenden die Bereit- Märkte und Gütertransportwege und leitet den wirtschaftli- schaft einfordern, sich auf anders geartete wissenschaftliche chen Zusammenbruch ein, und Fehleinschätzungen vor Ort Arbeitsweisen einzulassen; gleichzeitig werden sie aus den verzögern schnelle internationale Hilfe. jeweils anderen Fachbereichen vollkommen neue Forschungs- Aber auch bei Themen, die zunächst vor allem Fragen zur aspekte kennen lernen. Die Promotionsprojekte werden den Ausbreitung von Infektionskrankheiten in Europa betreffen, völlig neuen Qualitätsanspruch größtmöglicher Interdisziplina- ist eine fächerübergreifende Auseinandersetzung erforderlich. rität haben. Hierzu hat der Forschungsverbund vier wichtige Themen zur Für die Entwicklung nachhaltiger Ansätze zur Bekämpfung Verbreitung und Übertragung von Infektionskrankheiten des übertragbarer Erkrankungen unter Einbeziehung verschiedens- Menschen identifiziert: ter Aspekte und Vorrausetzungen bietet sich die Bündelung unterschiedlicher Expertisen aus Leibniz-Einrichtungen verschie- I. die Übertragung von schweren Infektionen wie TB und dener Ausrichtungen geradezu an. Zusammen können die HIV in Randzonen der Gesellschaft, die für die öffentliche biomedizinischen Fachzentren für Mikrobiologie und Virologie Gesundheitsversorgung schwer zugänglich sind, also illega- (BNI, FZB, HPI, HKI, DSMZ), Umwelt-, Klima- und Agrar-wissen- le Einwanderer, Obdachlose, Drogensüchtige, Prostituierte schaftlichen Fachzentren (ATB, IGB, IZW, PIK, TROPOS, ZALF, (MANtoMAN); ZMT) sowie die Fachzentren für Wirtschafts- und Sozialpolitik II. die Charakteristik der Ausbreitung von Krankheitserre- (GIGA, GESIS), die diesen Forschungsverbund ausmachen, ei- gern wie Grippeviren, Meningokokken oder Lungenentzün- nen radikal neuen Blick auf diese komplexe Thematik entwi- dungsbakterien über die Luft unter besonderen klimatischen ckeln. Frei von dem klassischen fachspezifischen Anspruch, Bedingungen im Freiland oder in Innenräumen wie Kranken- möglichst tief in die eigene Fachdisziplin einzusteigen, kann häusern oder Ställen (AIR); der Forschungsverbund über die ganze wissenschaftliche Brei- te seiner Partnerinstitute modular und skalierbar angelegte Zu- werden Workshops durchgeführt, um eine gemeinsame Ba- gangsweisen zur Problembewältigung erproben. sis zwischen den verschiedenen Disziplinen zu erschaffen und die unterschiedlichen wissenschaftlichen Arbeitsweisen ––––––––––––––––––– den jeweils anderen verständlich zu machen. Die Aktivitäten Social sciences Vectors ZALF, BNITM PIK, IGB, IZW UNI HH, FLI Water Krankheitserregern zur Verfügung zu stellen. aber auch die Chance zu beweisen, dass Wissenschaft in der Leibniz-Gemeinschaft auch anders und vor allem gemeinsam geht, dass Impact für die Menschheit jenseits von Impact-Faktoren der resultierenden Publikationen generiert werden kann, und dass theoria cum praxi vielfältig, originell und unerwartet Ma na HPI, ZMT, Public IZW, IGB relations ZALF, DSMZ Outbreak Perception an health sammeln, um sie den Leibnizinstituten zum Nachweis von Leibniz-Forschungsverbünde sind Beides, großes Wagnis Air Life sciences Enviromental science AFT, FZB GESIS, GIGA HPI, LSHTM PIK, TROPOS Graduate training Pathogen mit einbezogen. So können sie z. B. bundesweit Stechmücken se GESIS, GIGA FZB, BNITM Capacity building HPI, HKI Biomedicine Bürger werden in sogenannte „Citizen Science“ Projekte direkt Di s Man2Man Citizen science Hum sondern beteiligen auch die Öffentlichkeit. Bürgerinnen und Transmission me ge ion ct des Verbundes machen an den Toren der Institute nicht halt ea Inf e INFECTIONS ’21 erfolgreich sein kann. Dazu werden die beteiligten Institute nicht unerhebliche finanzielle und personelle Resourcen aus ihren eigenen Haushalten zur Verfügung stellen, um die zugesicherte Förderung zu ergänzen. Der LFV Infections ’21 bietet von der Thematik, der Methodik Ri sk und der Auswahl der beteiligten Institutionen beste Vorausset- ––––––––––––––––––– zungen, diese visionäre Programmatik mit Leben zu füllen. Es wird darauf ankommen, wirklich um die Ecke zu denken und Die Zusammenarbeit unterschiedlicher wissenschaftlicher die Vielfalt unserer Partner synergistisch zu nutzen – aber wo Disziplinen erfordert naturgemäß zunächst die gegenseitige sonst, wenn nicht bei Leibniz-Einrichtungen, ist dies möglich? Öffnung der jeweiligen Wissenschafts- und Sprach-Kultur: Das FZB koordiniert den LFV, weil wir uns über die strate- was für Mediziner Kohorten bzw. für Naturwissenschaftler gische Bedeutung dieser Verbünde für die Leibnizgemein- Experimente sind, sind für Sozialwissenschaftler Fragebögen schaft im Klaren sind: sie können Kernthemen bei zukünfti- und für Wirtschaftswissenschaftler Einkommensstatistiken – gen Evaluierungen sein, sie können zentrale Schwerpunkte die Kluft ist noch tief aber nicht unüberbrückbar, und es wird in Programmbudgets werden, sie können die Kriterien für viel Enthusiasmus und innere Überzeugung aller beteiligten Leibniz-orientierte Mittelvergaben der Zukunft werden. Das Wissenschaftlerinnen und Wissenschaftler brauchen, um FZB gestaltet so auch die Zukunft der Leibniz-Gemeinschaft hier nicht nur einen Brückenschlag zu bewerkstelligen. Dazu aktiv mit. LEIBNIZ-FORSC HUNGSVERBÜNDE – WARUM? Der Wissenschaftsrat hatte in seinem Gutachten „Perspektiven des deutschen Wissenschaftssystems“ angemahnt, dass die Leibniz-Gemeinschaft gerade aufgrund der Vielseitigkeit der wissenschaftlichen Disziplinen der ihr angehörenden Institute dazu in der Lage sein sollte, einen neuen Markenkern zu entwickeln. Dieser müsse das Credo theoria cum praxi durch eine transdisziplinäre Beantwortung brennender Fragen noch überzeugender als bisher verkörpern. Forschungsverbünde mit mehr als 10 Leibniz-Instituten sollen an den Grenzbereichen zwischen naturwissenschaftlichen, biomedizinischen, sozial- und wirtschaftswissenschaftlichen Forschungsfeldern Schnittmengen identifizieren, um sie für die Erarbeitung innovativer Lösungen für komplexe, gesellschaftlich relevante Fragestellungen zu nutzen. Es gibt zurzeit 11 Forschungsverbünde, wovon zwei finanziell von der Leibniz-Gemeinschaft über die nächsten 4 Jahre projektbezogen gefördert werden: Historische Authentizität und Infections ’21. Das FZB beteiligt sich auch an den LFV Medizintechnik, Wirkstoffe, Nanosicherheit und Biodiversität. Seite 11 / Presse-Highlights FORSCHUNGSZENTRUM BORSTEL 15 Millionen Euro für Laborumbau FORSCHUNGSZENTRUM BORSTEL für Ausbildung ausgezeichnet Stormaner Tageblatt, 19.12.2014 Foto: Olbertz Stormaner Tageblatt, 17.06.2013 ––––––––––––––– ––––––––––––––– EINE BÖSE ALTE BEKANNTE ––––––––––––––– Süddeutsche Zeitung, 23.04.2013 Foto: CDC GUTE TEILCHEN, SCHLECHTE TEILCHEN Welche Nanopartikel sind für die Lunge gefährlich? ––––––––––––––– BORSTEL Mit Kaugummi gegen Tuberkulose 125.10.2013, Deutschlandfunk ––––––––––––––– Lübecker Nachrichten, 09.07.2013 Foto: C. Steinhäuser GLUTEN IM ESSEN? Ein Neuer Schnelltest soll helfen ––––––––––––––– TUBERKULOSE Langsam gefährlich Hamburger Abendblatt, 12.11.2013 Foto: Marcelo Hernandez ––––––––––––––– 19.11.2013, DeutschlandRadio Wissen HIGHLIGHTS 2013/14 ––––––––––––––––––– ––––––––––––––––––– Ob im Fernsehen, im Radio oder in der Zeitung: In den Jahren 2013 und 2014 war das FZB häufig in den Medien vertreten – und das nicht nur zu wissenschaftlichen Themen. BESTE IM LANDE: Topnoten für Malina Schulz. Tangstedterin ist beste Auszubildende in SchleswigHolstein/21-Jährige mit Leibnizpreis ausgezeichnet ––––––––––––––– Stormarner Tageblatt, 9. November 2013 ; Foto: st GENOM-DETEKTIVE Was kann Bioinformatik gegen Keime ausrichten? ––––––––––––––– WAS HILFT BEI EINER POLLENALLERGIE? Arte, Wissenschaftsmagazin X:enius ––––––––––––––– 08.10.2013, NDR Visite BLUTVERGIFTUNG Wie erkennt und behandelt man sie? ––––––––––––––– 14. August 2014 Arte, Wissenschaftsmagazin X:enius & Schleswig-Holstein Magazin, 24.09.2013, 19:30 Uhr ABENTEUER DIAGNOSE: Unter Druck ––––––––––––––– NDR, 27.08.2013, 20:15 Uhr BAUMSTARKE AKTION: 57 Linden für die Allee gespendet ––––––––––––––– TUBERKULOSE-FORSCHUNG IN BORSTEL ––––––––––––––– FORSCHER GEHEN MIT GEN-TESTS GEGEN TUBERKULOSE VOR Schleswig-Holstein Magazin – 14.11.2014 ––––––––––––––– Focus, 12.02.2013 Foto: dpa/Angelika Warmuth WENN DAS HANDY DIE PATIENTEN ÜBER TUBERKULOSE AUFKLÄRT ––––––––––––––– FORSCHUNGSZENTRUM BORSTEL hilft Tbc-Kranken in Moldawien Lübecker Nachrichten, 30.04.2014 Foto: M. Hollinde 08.01.2014, Hamburger Abendblatt ––––––––––––––– 30.10.2014, Stormaner Tageblatt Seite 13 / Reportage ––––––––––––––––––– Autoren: Bettina Brand, Ulrike Schroer und Britta Weller Bilder: ©B. Weller, B. Brand, FZB –––––––––––––––––––– S PA S S I M J O B ––––––––––––––––––– Zufriedene Beschäftigte sind essenziell für den Erfolg. Viele Arbeitgeber setzen auf Well-Being-Programme. Z ufriedene Menschen sind in der Arbeitswelt produkti- •Soziale Bedingungen – Kontaktmöglichkeiten ver und sorgen so dafür, dass sich der Erfolg ihres Ar- •Finanzielle Bedingungen – Lohn, Sozialleistungen beitgebers mehrt. Dabei spielt es eine wichtige Rolle, dass Vorgesetzte sich mit ihren Mitarbeitern und Mitarbeiterin- Dabei ist eine Überforderung, wie auch eine Unterforde- nen auseinandersetzen, ihnen eine Vision bieten und die rung, im Arbeitsprozess zu vermeiden. Zudem stellen Burn- Möglichkeit ihre Potenziale zu entfalten, etwas zu lernen, out, aber auch Bore-out Phänomene unter Arbeitnehmer/ beizutragen, zu steuern oder bewirken zu können – schlicht innen einen hohen Kostenfaktor dar. die Sinnhaftigkeit ihrer eigenen Arbeit zu erkennen. Die Ansprüche der Beschäftigten an ihre Arbeit beziehen Das Forschungszentrum Borstel hat Mitarbeiterzufriedenheit als ein strategisches Ziel definiert. Das Zentrum sich laut Semmer und Udris* auf die Aspekte: befindet sich seit einigen Jahren in dem Prozess die Kom- •Arbeitsinhalt – Ganzheitlichkeit, Abwechslungsreichtum ponenten zu identifizieren, die für ein Borstel-spezifisches •Arbeitsbedingungen – Arbeitszeit, Belastungsfaktoren Well-Being-Paket zusammengetragen und welche Konse- •Organisationale Rahmenbedingungen – Sicherheit quenzen und Aktionen daraus abgeleitet werden müssen. des Arbeitsplatzes, Aufstiegschancen Dazu gehört der Wandel von der Präsenzkultur hin zu mehr * Bedeutung und Wirkung von Arbeit (Semmer und Udris in Schuler, 2007) Flexibilität und Ergebnisorientierung, eine Kultur der Ge- WAS IST PASSIERT? sundheit und des Wohlbefindens am Arbeitsplatz zu entwi- „Der größte Fehler ist, dass im Anschluss nichts passiert“, ckeln und die Vereinbarkeit von Beruf und Privatleben zu sagt der Wirtschaftspsychologe Felix Brodbeck. „Mitarbei- ermöglichen. terbefragungen sind kein Wunschkonzert“, warnt er. „Aber ein oder zwei Initiativen sollten daraus entstehen.“ Das FZB hat sich diese Warnung zu Herzen genommen und INSTRUMENT MITARBEITERBEFRAGUNG Um die einzelnen Komponenten eines Well-Being Pakets in den letzten zwei Jahren bestehende Angebote mit weite- identifizieren zu können, hat das FZB 2012 sich des Instru- ren Infrastukturen und zusätzlichen Maßnahmen ergänzt. ments einer Mitarbeiterbefragung bedient. Dazu wurde mit Die Graphik gibt einen Überlick über die bisherigen Angebote Prof. Albert Martin und Dr. Thorsten Jochims aus dem Insti- und die neu hinzugekommenen (kursiv). tut für Mittelstandsforschung, Universität Lüneburg, ein Befragungsbogen konzipiert und anschließend wissenschaft- NÄCHSTE SCHRITTE! lich ausgewertet. Bei einer Teilnehmerquote von Ø 57 % Die Entwicklung einer Kultur des Wohlbefindens und der Ge- ergab sich, dass allen vier Hauptbereichen der Befragung sundheit und der damit verbundenen Produktivität zu be- – Personalpolitik – Arbeitsgestaltung – Führungsverhalten - fördern ist ein dynamischer Prozess. An welcher Stelle des Arbeitgeber-Arbeitnehmer-Beziehung – eine gleichermaßen Prozesses wir uns zum jetzigen Zeitpunkt befinden, messen hohe Bedeutung für die Beschäftigten zukommt. Dabei kor- wir erneut im Frühjahr 2015 durch eine weitere Mitarbei- relierten Faktoren wie Verhaltensweisen der Vorgesetzten terbefragung. Sie wird uns Hinweise zum bisher Erreichten, (Hilfe, Interesse usw.), Aspekte der Mitwirkung (Informati- aber auch zu neuen Zielen und Aufgaben geben. Und auch on, Partizipation) und Probleme der Arbeitsorganisation danach werden wir mehr als die geforderten „ein bis zwei (Arbeitsverteilung, Festlegung der Arbeitszeiten) besonders Initiativen“ daraus entstehen lassen. stark mit der Arbeitszufriedenheit. ERFOLG KÖNNEN WIR NUR GEMEINSAM! FAZIT Die Mitarbeiterinnen, Mitarbeiter und Vorgesetzten des FZB sind insbesondere mit den Inhalten ihrer Arbeit sehr zufrieden. Sie arbeiten mit großem Engagement und sind bereit, sich stark einzusetzen. Die deutliche Mehrheit der Mitarbeiterinnen, Mitarbeiter und Vorgesetzte berichtete außerdem über eine solide Grundzufriedenheit und eine große Verbundenheit mit ihrem Arbeitgeber. Verbesserungsbedarf wurde insbesondere in den Bereichen des Führungsverhaltens, der Informationspolitik, der Arbeitsorganisation und in der Bereitstellung längerfristiger Beschäftigungsperspektiven gesehen. ––––––––––––––––––– DIE SOZIALREFERENTINNEN •haben das offene Ohr für alle sozialen Themen der Beschäftigten. •vermitteln Informationen zur Vereinbarkeit von Beruf und Familie (Was stellt mein Arbeitgeber bereits zur Verfügung? Was gibt es im externen Umfeld?) •nehmen Anregungen auf und geben sie an Entscheider weiter (Was wird noch gebraucht? Was sollte mein Arbeitgeber vorhalten?) •verstehen sich als Lotsinnen zu internen und externen Hilfeangeboten SIND ANSPRECHBAR, EMPATHISCH UND DISKRET. ––––––––––––––––––– Intranet (Folien aus FG-LeiterVersammlung im Intranet) Zentrumsdirektor – pers. Sprechzeiten Sozialreferate Beschwerdestelle GWP-Training Sprachkurse Betriebssport Retreats Betriebsfeiern Sicherheit (Neuorganisation) Betriebsärztlicher Dienst Gestaltung Kantine beratende Dienstleistung (Coaching/Supervision) Priorisierung von Entfristungen Gesicherte Finanzierung der Doktorarbeiten und der Anschlussphase 01 Gleichstellungsplan Training für Führungskräfte strukturierte Mitarbeitergespräche 03 Persönliche Ansprechpartner Sprachkurse Ausflugs- und Einkaufstouren niedrigschwellige Kommunikation Team-Events/ Mannschaftsgeist 05 Arbeitsplatz 02 Führungsverhalten 04 Integration ausl. Kollegen/innen Kinderbetreuung (Campus-Kita-Aufn. ab 9. Lebenswoche) flexible Arbeitszeiten Homeoffice Eltern-Kind-Büro Audit berufundfamilie Vermittlung von Engpassbetreuung Unterstützung bei der Eldercare Bedarfsgerechte Stundenreduzierung Beruf, Familie, Privatleben Jobsharing etc. 06 Seite 15 / Reportage Jürgen Nagel, Elektroinstallateur am Forschungszentrum Borstel I n Borstel herrscht ein sehr freundlicher Umgangston und eine nette Arbeitsatmosphäre – die Kollegen und Kolleginnen sind sehr hilfsbereit und hier wird auch mal Danke gesagt! Als ge- lernter Elektroinstallateur hat man hier ein breites und vielfältiges Aufgabengebiet – von kleinen Reparaturen bis hin zu den medizinischen Großgeräten. Das gibt es nicht überall! Lisa-Marie Johannssen Auszubildende Biologielaborantin M eine Ausbildung zur Biologielaborantin in Borstel ist sehr interessant und vielseitig. In den verschiedenen Forschungsgruppen lerne ich bereits neueste Methoden kennen und komme so in Kontakt mit aktuellen Themen in der Forschung. Die Qualität der Ausbildung ist sehr gut und über die Grenzen Schleswig-Holsteins bekannt, und ich errechne mir damit gute Chancen später auf dem Arbeitsmarkt. Dr. Christian Herzmann, Oberarzt an der Medizinischen Klinik I ch arbeite in Borstel, weil ich die Mischung aus Infektiologie, Intensivmedizin und anderen Lungenkrankheiten faszinierend finde. Die Medizinische Klinik bietet mir vielfältige Möglichkeiten, sich einzubringen – in der Funktionsdiagnostik, in der Forschung oder in der Patientenversorgung. Aufgrund der überschaubaren Größe der Abteilungen kenne ich fast jeden Mitarbeiter im Haus. Das vereinfacht vieles. Martina Ott Pflegekraft an der Medizinischen Klinik I ch arbeite gerne in Borstel, weil hier die Zusammenarbeit und das Arbeitsklima zwischen Ärzten und Pflegern stimmt und die Arbeit auf einer Intensivstation spannend und verantwortungsvoll ist. Weitere Vorteile sind, dass man im Öffentlichen Dienst arbeitet und Familie und Beruf gut unter einen Hut bekommt – für diese Vereinbarkeit wird hier in Borstel viel getan! Gudrun Lehwark-Yvetot Gästebetreuerin D ie Betreuung der ausländischen (Gast)Wissenschaftler wie wir sie verstehen und umsetzen, ermöglicht, auch außerhalb des Wissenschaftsbetriebes, ein besseres Zurechtfinden und eine gute Integration im Land. Diese wiederum ist Basis für gute wissenschaftliche Arbeit, nicht nur hier bei uns, sondern auch weltweit. Außerdem ist Kulturaustausch immer eine Bereicherung. Indem wir interkulturelle Kompetenzen auf- und ausbauen, leisten auch wir einen Beitrag zu Völkerverständigung und Toleranz untereinander. Es könnte nicht besser zum Thema dieser Tage passen. –––––––––––––––––––– Autor: Peter Zabel Autor: Ulrich Schaible Bild: © Uwe Groenewold, Lübeck, ©vege_fotolia, FZB –––––––––––––––––––– SABINE RÜSCH-GERDES ––––––––––––––––––– Ein Arbeitsleben für den Kampf gegen die Tuberkulose K aum jemand hat das Renomeé und die Außenwahrneh- mit ihrem Team trainiert und so die Basis für eine nachhal- mung unseres Institutes so sehr geprägt und gefördert tige Verbesserung der TB-Diagnostik in den Heimatländern wie Dr. Sabine Rüsch-Gerdes, die im Oktober diesen Jahres gelegt. Für diese außergewöhnlichen Initiativen und Ver- als langjährige Leiterin der Mykobakteriologie und des Na- dienste wurde Sabine Rüsch-Gerdes 2004 der international tionalen Referenzzentrums in den Ruhestand gegangen ist: renommierte Gardener-Middlebrook Award und 2005 der Ihre Tätigkeit begann 1977 am Forschungszentrum Bors- Verdienstorden der Bundesrepublik Deutschland verliehen. tel zunächst als stellvertretende Leiterin der Mykobakterio- Ihre einzigartige Kompetenz in der TB-Diagnostik macht Sa- logie und nach der Ernennung zum Nationalen Referenz- bine Rüsch-Gerdes zu einem gefragten Consultant interna- zentrum (NRZ) wurde Sabine Rüsch-Gerdes ab 1993 die tionaler Organisationen wie WHO, Ärzte ohne Grenzen und Leiterin des NRZ und seit 1995 Leiterin eines supranatio- dem internationalen Komiteé des Roten Kreuzes. nalen Referenzzentrums (SRL) der WHO. Zu ihren Verdiens- Sabine Rüsch-Gerdes hat in ihrer Laufbahn über 180 ten im weltweiten Kampf gegen die Tuberkulose zählen wissenschaftliche Publikationen veröffentlicht in teilweise die Evaluierungen der diagnostischen Meilensteine in der höchst angesehenen Journalen, so dass sie deutschland- ––––––––––––––––––– weit zu den am häufigsten zitierten Wissenschaftlern im Bereich der Mikrobiologie gehört. Aber auch für Interviews in Presse, Funk und Fernsehen ist sie eine höchst begehrte Partnerin und versteht wie kaum eine andere für ihre Sache, den Kampf gegen die Tuberkulose, klare und verständliche Botschaften zu senden. Herausragend ist ihre Kompetenz in der Beratung zu Fragen der TB-Diagnostik und ihre für Ratsuchende fast grenzenlose Erreichbarkeit. Sabine Rüsch-Gerdes hat in den Jahren in Borstel ein fantastisches Team geformt und ist mit vielen Forschergruppen des Instituts und der Medizinischen Klinik Borstel in höchst erfolgreichen Kooperationen auch freundschaftlich verbunden, so dass wir das Farewell an Sabine mit höchsten Respekt aber auch einer großen Portion Wehmut sagen. ––––––––––––––––––– Tuberkulosediagnostik (BACTEC, MGIT, Xpert) und die Qualitätssicherung dieser Methoden in 30 europäischen Ländern gefördert durch die ECDC. Seit 2010 ist sie Sprecherin aller SRLs der WHO und dieser Funktion Mitglied der Global Laboratory Initiative (GLI) und seit 2012 der European TB Laboratory Initiative der WHO (ELI). Auf ungezählten Reisen in die europäischen Länder aber auch in die Brennpunktländer Asiens und Afrikas hat Sabine Rüsch-Gerdes unermüdlich die Vor-Ort-Infrastruktur der TB-Diagnostik verbessert. Laborpersonal aus vielen Ländern weltweit wurde in Borstel Seite 17 / Farewell JABBAR AHMED ––––––––––––––––––– After 22 years at the Research Center Borstel (RCB), Prof. Jabbar Ahmed retired from his long-term engagement in December 2014. H is official date of retirement was already beginning of theileriosis, Jabbar was always dedicated to improve farm 2013 but he continued as supervisor of his residual animal health and thereby the situation of farmers. Conse- group due to a severe illness of his long-time co-worker, Ulrike quently, he translated his basic research in cell biology and Seitzer, who sadly passed away in May 2014. immunology to develop novel diagnostic tools and vaccines. Jabbar Sabir Ahmed, a graduate from Bagdad University Jabbar‘s group significantly contributed to the identification with a PhD in Veterinary Medicine from Bukarest Universi- of immunologically relevant Theileria proteins to be used as ty, joined the RCB in 1992 as head of the research group diagnostic tools but also developed a live attenuated vaccine Veterinary Infection Biology and Immunology. As veterinari- against the theileriosis. an, Jabbar was responsible for the animal facility and the Jabbar is a leading scientist in the field parasitology and Borstel training school for lab technicians till 2002. In his especially, Theileria research. With only few groups worldwi- various functions, Jabbar always assumed responsibilities de able to work with this complex infection, his group beca- for the Center. me quickly internationally recognized. He and his co-workers ––––––––––––––––––– contributed to more than 130 peer reviewed publications in the field. Always an excellent networker, Jabbar built up international networks in order to link European scientists with colleagues in Africa, the Middle East, China, Central and South-East Asia. This was made possible by Jabbar‘s excellent research instrumental for acquisition of a large body of grants. Beneath several DFG projects including one within the German-AfricaInitiative, he and his co-workers were also able to gain a significant number of EU grants including a highly acclaimed Marie-Curie Training Network. Jabbar was involved in 13 EUfunded projects within Frame Work Programs 5, 6, and 7. He coordinated three projects within FP6, one project in FP7 and functioned as Deputy-Coordinator in several other projects. Thereby, he acquired more than 2.5 mill of funding only to the RCB in the last 10 years. He also served as an evaluator for the European Commission and still continues to do so. As active teacher and mentor, Jabbar supervised more than ––––––––––––––––––– 30 students and led them to successful doctoral theses. Jabbar‘s prime scientific interest was to understand patho- He was involved in teaching at the University of Lübeck and genesis and cell biology of the tick-borne apicomplexa para- the Free University in Berlin, where he is adjunct Professor sites of the genus Theileria. These important ruminant patho- since 1993. gens cause of massive annual loss in farm animals primarily We wholeheartedly thank Jabbar for his long-term commit- in Africa and Asia. Together with his excellent research team ment to the RCB. We wish Jabbar a long lasting, rich and hap- and numerous international collaborators he dug deep into py retirement. the signal pathways and virulence properties abused by Theileria to reprogram lymphocytes to serve as host cells. As descendent of the Middle East, a region affected by ––––––––––––––––––––– Autoren: Barbara Kalsdorf and Christoph Lange Autoren: Bettina Brand and Stefan Ehlers Bild: ©Sagittaria, fotolia, FZB ––––––––––––––––––––– MARTIN ERNST ––––––––––––––––––– In March 2013, after 32 years of committed successful work, Dr. rer. nat. Martin Ernst retired from the Research Center Borstel. H aving studied Physics at the University of Stuttgart, of Immune Cell-Analytics) at the Research Center Borstel, Martin Ernst continued with studies in Biology at the Leibniz Center for Medicine and Biosciences. University Konstanz, where he completed his PhD thesis on Martin Ernst´s research activities focused on the func- „Passive cationtransport in normal and transformed mouse tion of human peripheral blood monocytes, macrophages fibroblasts“ in 1977. He continued to work as a postdoc at and other phagocytes in innate and adaptive immunity. He ––––––––––––––––––– did pioneering work to establish the fluorescence activated cell sorting, phagocytosis induced chemiluminescence and blood cell elutriation at the Research Center Borstel, which brought an essential contribution for all research departments of the center until now. These techniques enabled his group to identify four new monocyte subpopulations by their CD16 and CD64-expression pattern. He translated basic research to clinical questions and characterized blood and bronchoalveolar lavage cells of patients with various infectious diseases, rheumatic diseases, immune deficiencies or allergies. He also explored the impact of sleep on vaccination-induced specific memory T-cell function. Martin Ernst‘s special interests were immune cell interactions between the human hosts and mycobacteria, especially M. tuberculosis. His contributions have been substantial for the development of a novel immunospot-assay for the rapid diagnosis of smear-negative pulmonary tuberculosis which is the most sensitive diagnostic method used in clinical practice until today. Martin Ernst encouraged and lived the „Borstel spirit“ ever since he joined the faculty. He soon became one of the most respected colleagues on the campus. Martin’s excellent mentorship, endurance and accuracy has been instrumental for our own development and the development of many other Borstelanians. Beyond national borders he sustained a long lasting collaboration with international colleagues, e.g. from the Polish Universities of Krakow and Wroclaw. Martin provided an example ––––––––––––––––––– of the importance of collaboration and reciprocal stimulation between clinicians and basic scientific researchers. With his commitment and support for others he set standards. the Max-Planck-Institute for Immunobiology in Freiburg and We wholeheartedly thank Martin Ernst for his dedication followed the new Director Hans-Dieter Flad in 1981 to Nort- and support. Although he is now retired, we still have the hern Germany, where Martin became the Head of the Divisi- pleasure to meet him occasionally and to receive his advice. on of Cell Biology (which in 2004 was re-named to Division All best wishes shall be with him for his future steps. Seite 19 / Farewell HELMUT HAAS ––––––––––––––––––– In September 2013, Privatdozent Dr. med. Helmut Haas retired from his long-term engagement at the Research Center Borstel. I n 1981, Helmut started his work on the molecular charac- lized in high-throughput drug screening on in vitro-cultured terization of grass pollen allergens via immunoblotting in schistosomes and production of immunomodulatory parasite the Research Group ‘Clinical Immunology’. 1987 saw Helmut material (www.helminguard.de; user name ‘gast’, password setting up his own research group (Cellular Allergology), focu- ‘helminguard’). sing on the regulation of IgE synthesis. In 1990 he took a sabbatical and visited the lab of Prof. Kimishige Ishizaka, La Jolla ––––––––––––––––––– Institute for Allergy and Immunology, California. This visit had significant long-term effects and inspired him to “learn from the worm” (Schistosoma mansoni) in terms of efficiently activating or dampening immune defense reactions: Helmut became a bona-fide “worm whisperer”. In the next years Helmut and his coworkers established in vitro systems for studying mechanisms of Th2 and IgE induction including preparation of highly purified human basophils. They were among the first to elucidate the role of basophils as a source of early IL4, and he and his group remain to this day one of the very few experts in human basophils (which, Helmut will not cease to tell you, are very different from mouse basophils!) Supported by the Federal Ministry of Education and Science and the EU Consortium ‘TheSchistoVac’ the Haas team developed a unique technique to replicate the schistosomes’ natural habitat thereby enabling the examination of the development of the helminthes from larvae to adulthood, including the pairing and deposition of eggs. Working in collaboration with the teams around Mike J. Doenhoff (Nottingham), Maria Yazdanbakhsh (Leiden), Christoph G. Grevelding (Giessen) and other groups Helmut and his coworkers identified three key molecules from Schistosoma eggs that may help to develop immunomodulatory, and in particular anti-inflammatory drugs in the future: Omega-1, IL-4-inducing principle of Schistosoma mansoni eggs (IPSE) / alpha-1 and kappa-5. Helmut Haas was also an active teacher and mentor. He supervised many students and led them to successful doctoral theses and lectured at the University of Lübeck as well as at the Borstel Biomedical Research School. His dry humour and his thoughtful reflections on the philosophy of scientific exploration will be well remembered as a unique part of Borstel culture. Today, although officially retired, Helmut Haas is often still on campus. He founded helminGuard, a company specia- ––––––––––––––––––– Helmut, we thank you for your contributions which were often surprising and always innovative. We wish you all the best for the years to come! ––––––––––––––––––– ––––––––––––––––––– BMBF finanzierter GLUTEVIS Verbund entwickelt Nachweis- REACH-4-Moldova“ – DAAD fördert die Partnerschaft zwi- methoden zur Identifizierung glutenhaltiger Lebensmittel; schen Lübeck-Borstel-Moldawien Koordination Andreas und Barbara Frey, FG Mukosale Immunologie und Diagnostik REACH-4-Moldova“ – DAAD fosters Lübeck-Borstel-Moldova partnership Federal Ministry of Education and Science supports GLUTEVIS network in developing detection methods for the identification of gluten-containing foods. Coordination ––––––––––––––––––– Innovationspreis 2013 der BioRegionen Deutschland geht Andreas und Barbara Frey, RG Mucosal Immunology an Klaus Brandenburg (FG Biophysik) für die Entwicklung and Diagnostics eines hochwirksamen, neuartigen Anti-Sepsis Medikaments ––––––––––––––––––– Innovation Award 2013 awarded by BioRegion Germany Volkswagenstiftung fördert Entwicklung innovativer TB Dia- goes to Klaus Brandenburg (RG Biophysics) for the deve- gnostik; PI Christian Herzmann, FG Klinische Infektiologie lopment of a highly efficient, novel sepsis treatment. Volkswagenstiftung promotes the design of innovative TB diagnostic tools; PI Christian Herzmann, RG Clinical Infectious Diseases ––––––––––––––––––– ––––––––––––––––––– Den Deutscher Ideenpreis 2013 erhält Christian Herzmann (FG Klinische Infektiologie) für die Smartphone-basierte Tuberkulose Aufklärung ‚Explain TB!‘ in über 50 Sprachen Walter-Dosch-Lehrpreis der Universität Lübeck für Christian Herzmann (RG Clinical Infectious Diseases) hervorragende Lehre nicht-professoraler Dozenten geht receives the German Idea Award 2013 for a smart-phone an Uwe Mamat, FG Strukturbiochemie based mobile tuberculosis education for patients and health care workers. Available in more than 50 languages Uwe Mamat receives the Walter-Dosch-Lehrpreis, University Lübeck, for excellent teaching, RG Structural Biochemistry ––––––––––––––––––– ––––––––––––––––––– Der IHK Ausbildungs-Award, Schleswig-Holstein, für herausragende Leistungen in der betrieblichen Ausbildung geht an Susanne Zähringer & Team Sina Webering erhält den Young Investigator Award der European Respiratory Society, FG Experimentelle The chamber of Industry and Commerce, Schleswig- Pneumologie Holstein, awarded the excellent in-firm training offered by Susanne Zähringer and her team with the The Young Investigator Award awarded by the European Respiratory Society goes to Sina Webering, RG Experimental Pneumology ––––––––––––––––––– Training Award 2013 ––––––––––––––––––– GBM Innovation Award for Young Scientists geht an Christine Steinhäuser, FG Mikrobielle Grenzflächenbiologie Arte / X:enius begibt sich mit der Kamera auf die Spur der Genom-Detektive, FG Molekulare Mykobakteriologie GBM Innovation Award for Young Scientists goes to Christine Steinhäuser, RG Microbial Interface Biology Arte / Xenius tracking the Genome Detectives in front of a live camera, RG Molecular Mycobacteriology ––––––––––––––––––– ––––––––––––––––––– Seite 21 / Highlights ––––––––––––––––––– ––––––––––––––––––– Malina Schulz – 3-fache Gewinnerin: beste Auszubildende Der FZB Förderverein VEIAP zeichnet Viola Schleusener, in Schleswig-Holstein, im Bundesgebiet und in der Leibniz- FG Bioinformatik, für ihre hervorragende Masterarbeit aus Gemeinschaft The Förderverein (supporters‘ association) VEIAP honored Three times winner: best trainee of Schleswig-Holstein Viola Schleusener, RG Bioinformatics, for her excellent and nationwide and winner of the Leibniz-Auszubildenden scientific work (MSc) Preis – Malina Schulz ––––––––––––––––––– ––––––––––––––––––– Der Promotionspreis des Kreises Segeberg geht an Lars Der CleanColiTM Kit zur Produktion LPS freier Bakterien, Lunding, FG Mausmodelle des Asthma, für die beste entwickelt von Uwe Mamat und Kathleen Wilke, FG Struk- Doktorarbeit 2013 turbiochemie) wird unter die Top Ten der besten Innovationen in der Wissenschaftstechnologie gewählt. The prize for the best thesis 2013 awarded by the district Segeberg goes to Lars Lunding, Bacteria devoid of LPS (CleanColiTM) now ready for industrial application (Uwe Mamat, Kathleen Wilke, RG Structural Biochemistry), New Scientist: ‚Top10 Innovations in RG Asthma Mouse Models. ––––––––––––––––––– Science Technology’ ––––––––––––––––––– Publikation Blockbuster: PLOS Medicine (FG Mol. Mykobakteriologie), Nature Genetics (FG Mol. Mykobakteriologie); Nature Communications (FG Strukturbiochemie), Nature Chemical Biology (FG Biophysik), Gastroenterology (FG Strukturbiochemie, FG Biochemische Immunologie) Publication Blockbusters: PLOS Medicine (RG Mol. Mycobacteriology), Nature Genetics (RG Mol. Mycobacteriology); Nature Communications (RG Structural Biochemistry), Nature Chemical Biology(RG Biophysics), Gastroenterology (RG Structural Biochemistry, RG Biochemical Immunology) ––––––––––––––––––– HIGHLIGHTS ––––––––––––––––––– ––––––––––––––––––– ––––––––––––––––––– EU – NAREB (Nanotherapeutics for Antibiotic Resistant ––––––––––––––––––– Susanne Krauss-Etschmann tritt am 1. Oktober 2014 die Emerging Bacterial pathogens): das Konsortium kämpft W3-Professur „Experimentelle Asthmaforschung“ am FZB gegen multiresistente Mykobakterien und Methicillin-re- an. Die Berufung dieser Professur erfolgte gemeinsam mit sistente Staphylokokken. Beide Erreger sind für schwerste der Christian-Albrechts-Universität zu Kiel mit dem Ziel, Erkrankungen verantwortlich, die eine Langzeitherapie und den Schwerpunkt „Disease Area Asthma and Allergy“ des eine Intensivbetreuung erfordern. Ulrich Schaible (Zell. Airway Research Center North (ARCN) und den Schwer- Mikrobiologie), Elvira Richter (Diagn. Mykobakteriologie) punkt „Kiel Life Science“ der CAU zu stärken. EU – NAREB (Nanotherapeutics for Antibiotic Resistant Susanne Krauss-Etschmann takes over her Professorship Emerging Bacterial pathogens): fight against Multi-Drug ‘Experimental Asthma Research’ on October 1st, 2014. Resistant (MDR) Mycobacterium tuberculosis (M. tubercu- The chair was endowed by the University of Kiel and the losis) (TB) and Methicillin Resistant Staphylococcus aureus FZB to foster the ‘Disease Area Asthma and Allergy’ within (MRSA), leading to serious diseases which usually require the German Center for Lung Research intensive care treatment with a long hospitalization. Ulrich Schaible (Cell. Microbiology), Elvira Richter (Diag. Mycobacteriology). ––––––––––––––––––– ––––––––––––––––––– NanoCOLT untersucht in einem deutschlandweiten Verbund, wie sich Kohlenstoff-Nanopartikel auf den Atmungsapparat auswirken. Das Bundesforschungsministerium Leibniz-Forschungsverbund ‚Infections 21‘ – Transmission gibt in den nächsten drei Jahren insgesamt zirka 2,4 Control of Infections in the 21st Century“ – zur Kontrolle, Millionen Euro für das Konsortium um die Langzeitwirkung Prävention und Bekämpfung von Infektionskrankheiten. An modifizierter Carbon Black Nanopartikel auf gesunde und dem transdisziplinären, von der Leibniz-Gemeinschaft ge- vorgeschädigte Lungen zu untersuchen. förderten Projekt sind 14 Leibniz-Institute und drei externe Partner beteiligt. Koordination Ulrich Schaible. NanoCOLT, a German consortium, is devoted to investiga- Leibniz-Research-Network ‚Infections 21‘ – Transmissi- human respiratory tract. The Federal Ministry of Education on Control of Infections in the 21st Century“ – control, and Science supports the project with 2.4 Mio. Euro. te the long-term effects of carbon-nano-particles on the prevention and combat of infectious diseases. The Leibniz Association-funded network consisting of 14 Leibniz insti- ––––––––––––––––––– tutions and three external partners pursues a transdiscipli- Zum 1. Juli 2014 hat Christoph Lange die W3-Professur nary approach. Coordination Ulrich Schaible. „International Health/Infectious Diseases“ angetreten, die ––––––––––––––––––– im Rahmen der Kooperation zwischen dem Forschungszentrum Borstel, der Universität zu Lübeck und dem Deut- Mit Schwarmintelligenz gegen Tuberkulose: Christoph schen Zentrum für Infektionsforschung (DZIF) eingerichtet Lange erhält den Wissenschaftspreis des Stifterverban- wurde. Christoph Lange wird das Klinische Tuberkulosezen- des. Er wird für seine herausragenden Anstrengungen im trum (CLinTB) des DZIFs am Forschungszentrum Kampf gegen die Ausbreitung hochgradig antibiotikaresis- Borstel leiten. tenter Tuberkulose-Erreger ausgezeichnet. Christoph Lange takes over the chair ‚International Using swam intelligence to fight multi-resistant tubercu- Health/ Infectious Diseases’ on July 1, 2014. The chair losis: Christoph Lange receives the prestigious Science was implemented by the FZB, the University of Lübeck Award of the Deutscher Stifterverband for his outstanding and the German Center for Infectious Diseases. Christoph work in combating the spread of highly resistant tuberculo- Lange will also be head of the Clinical Tuberculosis Center, sis pathogens. financed in part by the German Center for Infection Re- ––––––––––––––––––– search located at the FZB. ––––––––––––––––––– Seite 23 / Highlights ––––––––––––––––––– gezeichnet: Zum einen erhielt er den von der Deutschen Die Universität zu Lübeck hat gemeinsam mit dem For- Lungenstiftung ausgelobten Preis für die beste schungszentrum Borstel zum 1. Mai 2014 die neue W3 experimentelle Dissertation, zum anderen wurde sein Professur für Molekulare und Experimentelle Mykobakterio- wissenschaftlicher Vortrag als einer der vier besten logie im Rahmen des Deutschen Zentrums für Infektions- Kongressbeiträge ausgezeichnet. forschung (DZIF) eingerichtet. Besetzt wurde diese Stelle durch den Molekular-Epidemiologen und Populationsgene- Annual Meeting of the German Society for Pneumology tiker Stefan Niemann. and Ventilation Medicine: Lars Lunding, RG Asthma Mouse Models, receives two awards at once: one for the best Stefan Niemann takes over the chair ‚Molecular and Ex- experimental thesis and one for giving one of the four best perimental Mycobacteriology‘ on May 1st, 2014. The chair presentations. was founded by the University of Lübeck and the FZB. ––––––––––––––––––– ––––––––––––––––––– Promotionspreis Kreis Segeberg geht an Matthias Merker, Das Projekt „Explain TB“ (Christian Herzmann) des FZB hat FG Molekulare Mykobakteriologie, für seine mit summa den mit 15.000 Euro dotierten 1. Platz des Aspirin Sozial- cum laude ausgezeichnete Doktorarbeit. preises 2014 erreicht. The prize for the best thesis 2014 awarded by the district Aspirin Sozialpreis – First place for the project ‘Explain TB’ Segeberg goes to Matthias Merker, RG Molecular Myco- (Christian Herzmann) endowed with 15,000 Euro. bacteriology. His thesis was assessed ‘summa cum laude’. ––––––––––––––––––– Auf Tuchfühlung mit Nobelpreisträgern: Borsteler Nach- ––––––––––––––––––– Publikation Blockbuster: Nature (Mol. Mykobakteriologie), wuchswissenschaftler/innen Silke Feuerriegel (Mol. Nature Genetics (Mol. Mykobakteriologie), Plos Medicine Mykobakteriologie) und Michael Wegmann (Mausmodelle (Mol. Mykobakteriologie), Lancet (Mol. Mykobakteriologie), des Asthma) zum 64. Lindau Nobel Laureate Meeting American Journal of Respiratory and Critical Care Medicine eingeladen. (Klin. Infektiologie), Nature Communications (Strukturbiochemie) Young scientists in close contact with nobel laureates at the 64th Lindau Nobel Laureate Meeting: Silke Feuerriegel Publication Blockbusters: Nature (Mol. Mycobakteriology), (Mol. Mycobacteriology) and Michael Wegmann (Asthma Nature Genetics (Mol. Mycobacteriology), Plos Medicine Mouse Models). (Mol. Mycobacteriology), Lancet (Mol. Mycobacteriology), ––––––––––––––––––– Lars Lunding, FG Mausmodelle des Asthma, wurde auf dem Jahreskongress der Deutschen Gesellschaft für Pneumologie und Beatmungsmedizin gleich zweimal aus- American Journal of Respiratory and Critical Care Medicine (Clin. Infectious Diseases), Nature Communications (Structural Biochemistry) ––––––––––––––––––– HIGHLIGHTS ––––––––––––––––––– ––––––––––––––––––– ––––––––––––––––––––– Autoren: Bettina Brand, Kurt Fellenberg und Britta Weller Bilder: FZB ––––––––––––––––––––– Bringing bioinformatic expertise to the campus, March 2013. ––––––––––––––– KURT FELLENBERG Populationsgenetik von Tuberkulosebakterien. ––––––––––––––– STEFAN NIEMANN MENSCHEN•PEOPLE ––––––––––––––– 2013/2014 Disposition für Asthma und COPD. ––––––––––––––– SUSANNE KRAUSSETSCHMANN Passion for lipidomics. ––––––––––––––– Die großen Zwei: Malaria und Tuberkolose. DOMINIK SCHWUDKE ––––––––––––––– BIANCA SCHNEIDER Translationale Tuberkuloseforschung. ––––––––––––––– CHRISTOPH LANGE Seite 25 / Menschen • People B ioinformatics is a relatively new field of research. In ry networks are usually conserved. While this may appear 1998, after taking my diploma in biology, I had the op- theoretical, everyday work often deals with practical issues portunity to start my PhD in Klaus Rajewsky‘s lab, a group such as the question how to make a program user-friendly renowned for producing transgenic mice and Nature pa- or how to protect data uploaded by one user of a web in- pers. I declined in order to do bioinformatics, instead, and I terface from being accessed by any other user of the same have never regretted my choice. Specializing in quantitative web interface. Also, computers do not tend to be less un- high-throughput data, I worked on data stemming from al- relenting to bioinformaticians than to others. There are just most all microarray platforms including antibody chips, but more programs to be installed and operated. Thus, much also from 2D-DIGE gels, next generation sequencing data, like in the laboratory, most of the time is spent on „making quantitative LC-MS/MS mass spectrometry data (TMT6- things work“. plex, SILAC, and label-free) and other platforms. However, our work still differs from that of other resear- ––––––––––––––––––– chers in Borstel. A bioinformatician‘s „findings“ consist of Specialized in quantitative high-throughput data. end, software. If the software is any good, others may use ––––––––––––––––––– Comparing the impact of biomedical journals to the top The bioinformatician‘s work starts with wondering how to journals of my „trade“, bioinformatics appears irrelevant. store these data in a manner optimal for analysis („data But sometimes it provides its followers with a good laugh. warehousing“). Data are then visualized and systematically Starting in a new place (no, not Borstel), I once was provi- explored („data mining“). Additional information, e. g. cli- ded with new „hot“ data whose interpretation and publica- nical data further characterizing the patients or also gene tion was most pressing and, so I was informed with urgent ontology terms annotating the genes can be added in order warning, of utmost priority. What people involved usually do to interpret large datasets. Datasets are sometimes put into not know is that those files‘ headers always contain a time an even larger context of many other datasets, e.g. by meta- stamp recording exactly when the data have been acquired. analysis of expression data. Finally, high-throughput data These had been waiting for somebody to tackle their inter- provide the option to infer gene regulatory networks from pretation for more than two years. And, what is more, like them, opening the gate to systems biology. The so-called explorers discovering a new land, biological findings may be- „reverse engineering“ even works across species borders long to the people acquiring the data, but bioinformaticians because, much like gene sequences, important regulato- are often the first to see them. KURT FELLENBERG K e.g. database structures, methods, algorithms, or, in the it to process their data, thus producing important findings. oinfektionen treten natürlicherweise in Gebieten mit inwiefern sich diese negativ auf den Verlauf und die Kontrol- hoher Inzidenz verschiedener Infektionskrankheiten le einer Tuberkulose auswirkt und welche immunologischen auf. Während einer Koinfektion können die Immunantworten gegen die jeweiligen Erreger nachhaltig verändert werden, was nicht nur Krankheitsverlauf sondern auch Vak- Mechanismen dem zu Grunde liegen. Langfristig ist eine Ausweitung auf weitere relevante Koinfektionen geplant. zinierung, Diagnose und Therapie beeinträchtigen kann. Bianca Schneider hat an der Universität Bonn Biologie Bianca Schneider nutzt ihre Expertise in experimentellen studiert und dort 2006 mit summa cum laude promoviert. Mausmodellen, um die wechselseitigen Auswirkungen von Nach einer ersten PostDoc Zeit am Max-Planck Institut für Koinfektionen auf diese klinisch bedeutsamen Outcome- Infektionsbiologie in Berlin hat sie für vier Jahre zunächst Parameter zu untersuchen. als Visiting später als Postdoctoral Research Fellow an der ––––––––––––––––––– renommierten London School of Hygiene & Tropical Medici- BIANCA SCHNEIDER ne in London gearbeitet. 2011 kam sie an das FZB und war Bianca Schneider leitet seit dem 1. April 2014 die neu eingerichtete Nachwuchsgruppe „Koinfektion“ im Programmbereich Infektionen. bisher als wissenschaftliche Mitarbeiterin in der FG Zelluläre Mikrobiologie von Prof. Schaible tätig. Bianca Schnei- ––––––––––––––––––– der ist am Lehrprogramm des MSc Studiengangs ‚Infection Biology‘ und am Ausbildungsprogramm der Leibniz-Gradu- Gemeinsam mit ihrem Team wird Bianca Schneider in den iertenschule ‚Modellsysteme für Infektionskrankheiten‘ und nächsten 4 Jahren zunächst verschiedene Aspekte der Ko- der ‚Borstel Biomedical Research School (BBRS)‘ beteiligt. infektion mit Malaria charakterisieren, um herauszufinden, Zudem ist sie die Ombudsfrau der BBRS. T he group focuses on the identification and quantitation Dominik Schwudke studied chemistry at the Humboldt- of membrane components and immunological active University Berlin and graduated in the group of Prof. Lin- biomolecules and is specialized in the analysis of lipids, gly- scheid working in close cooperation with the group of Prof. co-conjugates and cell wall components. Dominik Schwudke Appel at the Robert-Koch-Institute (Berlin). In 2004 he did and his team aim to reveal signaling cascades and biosyn- his postdoctoral work in Andrej Shevchenko’s laboratory, thetic pathways on the molecular post-transcriptional level, MPI of Molecular Cell Biology and Genetics in Dresden, fol- which are altered due to inflammation and infection. For lowed by a long-term commitment (2009-2013) at the Nati- that they develop and apply OMICs workflows and methods onal Centre for Biological Sciences (TIFR) Bangalore (India) for structural characterization. They are experts in mass as principle investigator funded by the Wellcome Trust. In spectrometry (MS) and nuclear magnetic resonance (NMR) 2009 he received the NCBS-Merck & Co International Inves- approaches enabling in-depth studies of metabolic proces- tigator Award. ses in biological model system as well as clinical samples. Since 2013 Dominik Schwudke has been a member of the ––––––––––––––––––– Cluster of Excellence 306 ‘Inflammation at Interfaces’ and Dominik Schwudke joined the FZB in February 2013 as head of the Research Group Bioanalytical Chemistry. (German Center for Infection Research) as well as the Airway DOMINIK SCHWUDKE involved in the Thematic Translational Unit of Tuberculosis Research Center North (German Center for Lung Research). ––––––––––––––––––– D ie Berufung dieser Professur erfolgte gemeinsam mit dieser frühkindlichen Prägung könnte in Zukunft dazu füh- der Christian-Albrechts-Universität zu Kiel (CAU) mit ren, diese Erkrankungen bereits vor ihrer Entstehung positiv dem Ziel, den Schwerpunkt „Disease Area Asthma and Al- zu beeinflussen. Dafür könnte man Substanzen aus nicht- lergy“ des Airway Research Center North (ARCN) und den infektiösen Bakterien verwenden, die vorbeugend eingenom- Schwerpunkt „Kiel Life Science“ der CAU zu stärken. Zu- men werden. dem ist die Professur mit der Leitung der Forschungsgrup- Susanne Krauss-Etschmann hat in München und Paris Hu- pe „Experimentelle Asthmaforschung“ im Programmbereich manmedizin studiert und ist Fachimmunologin sowie Fach- Asthma & Allergie am FZB verbunden, die sich mit den vor- ärztin für Kinderheilkunde und Jugendmedizin. Im Jahr 2006 geburtlichen und frühkindlichen Ursprüngen chronischer habilitierte sie im Fach Pädiatrie zum Thema „Modulation Lungenerkrankungen beschäftigt. TH2-assoziierter Immunantworten im Kindesalter“. ––––––––––––––––––– SUSANNE KRAUSS-ETSCHMANN Susanne Krauss-Etschmann hat am 01.10.2014 die W3Professur „Experimentelle Asthmaforschung“ angetreten. Prof. Krauss-Etschmann war von 2010 bis 2014 im Comprehensive Pneumology Center (CPC), einem Kooperationsprojekt des Klinikums der Ludwig-Maximilians Universität, den Asklepios Fachkliniken und des Helmholtz Zentrums München ––––––––––––––––––– – Deutsches Forschungszentrum für Gesundheit und Umwelt Chronische Lungenerkrankungen stellen derzeit weltweit tätig, wo sie die Arbeitsgruppe „Developmental Origins of eine der häufigsten Todesursachen dar. Allein in Europa wer- Respiratory Disease“ leitete und sich mit Faktoren und den den dadurch derzeit jährlich ca. 200 Mrd Euro an Gesund- Mechanismen beschäftigte, die für eine frühe Prägung des heitskosten verursacht, wovon der Großteil auf Asthma und Asthmarisikos von Bedeutung sind. chronisch obstruktive Lungenerkrankungen entfällt. Studien Seit 2013 war sie als Principal Investigator (PI) am Deut- haben gezeigt, dass ein Zusammenhang zwischen Umwelt- schen Zentrum für Lungenforschung (DZL) am Standort Mün- faktoren, die in vorgeburtlichen und frühen Entwicklungs- chen (CPC-M) tätig. Die Rolle als PI wird sie auch am hiesigen fenstern wirksam sind und dem Risiko im späteren Leben DZL-Standort ARCN weiterführen, was einerseits innerhalb an einer chronischen Lungenerkrankung zu leiden. Einer des DZL zur Bündelung der tierexperimentellen Kompetenz der Hauptrisikofaktoren eines Kindes später Asthma oder in der Asthmaforschung am Standort ARCN führt und ande- COPD zu bekommen, ist mütterliches Rauchen während der rerseits zur weiteren Vernetzung der Standorte beiträgt. Sie Schwangerschaft. Die Gruppe um Susane Krauss-Etschmann leitet zudem seit 2012 das EU-geförderte BMBS COST Action möchte verstehen, wie die Krankheitsrisiken für Asthma und BM1201 Programm „Developmental Origins of Chronic Lung COPD während der Schwangerschaft und im frühen Kindesal- Disease“ - ein internationales Netzwerk, an dem Forscher ter geprägt werden. Entsprechend werden die Mechanismen aus 26 Ländern beteiligt sind und sich verschiedenen Aspek- der vorgeburtlichen tabakinduzierten Krankheitsprägung in ten der Entwicklungssprünge chronischer Lungenerkrankun- einem Mausmodell untersucht. Ein besseres Verständnis gen widmen. Seite 27 / People/Menschen Z um 01.07.2014 hat Christoph Lange die W3-Professur Immunrekonstitution bei der HIV-Infektion unter antiretroviraler „International Health/Infectious Diseases“ angetreten, Therapie und wurde 2009 zum außerordentlichen Professor an die im Rahmen der Kooperation zwischen dem Forschungs- der Universität zu Lübeck ernannt. Neben der Facharztanerken- zentrum Borstel, der Universität zu Lübeck und dem Deut- nung in Innerer Medizin und dem Gebiet Pneumologie verfügt schen Zentrum für Infektionsforschung (DZIF) eingerichtet er über die Zusatzqualifikationen in den Bereichen Infektiologie, wurde. Christoph Lange leitet das Klinische Tuberkulose- Intensivmedizin, Allergologie und Schlafmedizin. zentrum (CLinTB) des DZIFs am FZB. Den zeitgleichen Ruf Christoph Lange war Gründungspräsident des Tuberculosis auf eine Professur für Klinische Infektiologie am Karolinska Network European Trialsgroup und ist der Leiter der Respirato- Institut in Stockholm, Schweden, lehnte der klinische Tu- ry Infections Assembly der European Respiratory Society (ERS). berkulosespezialist mit den Forschungsschwerpunkten auf Neben der Mitarbeit in nationalen und internationalen Fach- den Gebieten der M/XDR-Tuberkulose und der Biomarker- gremien ist Christoph Lange stellvertretender Vorsitzender der entwicklung ab. Ethikkommission der Ärztekammer in Schleswig Holstein. ––––––––––––––––––– CHRISTOPH LANGE Für seine wissenschaftliche Leistungen und Beiträge zur translationalen Forschung auf dem Gebiet der Tuberkulose Wissenschaftspreis des Deutschen Stifterverbandes, 2014. erhielt Christoph Lange die Ehrendoktorwürde der Staatli- ––––––––––––––––––– temitanu“ (USMF) in Chisinau/Moldawien. Seit 2012 ist er chen Universität für Medizin und Pharmazie „Nicolae Tes- Nach Abschluss des Studiums der Biologie an den Univer- Gastprofessor der USMF und seit 2013 Gastprofessor der sitäten Kiel und Freiburg hat Professor Lange seine wissen- University of Namibia School of Medicine in Windhoek. Für schaftliche Ausbildung durch ein Medizinstudium an der die Universität Lübeck ist Christoph Lange federführend an Universität Witten/Herdeck ergänzt und 1994 erfolgreich den Partnerschaften mit den Universitäten in Moldawien abgeschlossen. Während seiner Assistenzarztzeit war er für und Namibia über DAAD-Pagel-Partnerprogramme beteiligt. 6 Monate an der University Cape Town (Südafrika) und für 2 2014 erhielt er für seine herausragenden Anstrengungen im Jahre an der Case Western Reserve University in Cleveland Kampf gegen die Ausbreitung hochgradig antibiotikaresisten- (Ohio, USA) klinisch und wissenschaftlich tätig. ter Tuberkulose-Erreger den Wissenschaftspreis des Deut- 2004 habilitierte er sich im Fach der Inneren Medizin über die D schen Stifterverbandes. ie Universität zu Lübeck hat gemeinsam mit dem For- erhielt eine Gastprofessur an der Staatlichen Universität für schungszentrum Borstel zum 01.05.2014 eine neue Medizin und Pharmazie „Nicolae Testemitanu“ in Moldawien. W3 Professur für Molekulare und Experimentelle Mykobakteriologie im Rahmen des Deutschen Zentrums für Infektionsforschung (DZIF) eingerichtet. Besetzt wurde diese Stelle durch den Molekular-Epidemiologen und Populationsgene- ––––––––––––––––––– STEFAN NIEMANN International anerkannter Genomdetektiv. tiker Stefan Niemann. Zu den zentralen Aufgabengebieten ––––––––––––––––––– gehören die Leitung der FG Molekulare Mykobakteriologie Stefan Niemann gehört weltweit zu den führenden Wis- am FZB, die Beteiligung am Nationalen Referenzzentrum für senschaftlern auf dem Gebiet der Populationsgenetik von Mykobakterien und die Vertretung des Bereiches Epidemio- Tuberkulosebakterien und hat in den letzten Jahren heraus- logie der Thematischen Translationseinheit „Tuberkulose“ ragende Forschungsbeiträge zur Entstehung und Verbrei- des DZIFs. Zusätzlich beteiligt sich Stefan Niemann künftig tung multiresistenter (MDR) und extrem resistenter (XDR) an der Graduiertenausbildung und der Lehre in den Studien- Tuberkulose-Erreger geleistet. 2011 erhielt er dafür den Eva gängen Molecular Life Sciences und Infection Biology an der und Klaus Grohe-Preis der Berlin-Brandenburgischen Aka- Universität zu Lübeck. demie der Wissenschaften. Stefan Niemann hat im Fach Biologie an der Universität Stefan Niemann ist Koordinator des EU-Projektes „Pa- Bielefeld promoviert und 2004 in den Bereichen Molekulare thoNgenTrace” und der Thematischen Translationseinheit Biologie und Mikrobiologie an der Universität zu Lübeck ha- „Tuberkulose“ des DZIFs. Seit 2010 ist er zudem Präsident bilitiert. Seit 2006 leitet er die FG Molekulare Mykobateriolo- der Europäischen Gesellschaft für Mykobakteriologie und gie und beschäftigt sich schwerpunktmäßig mit dem Thema Vorstand der Fachgruppe Mikrobielle Systematik, Popula- Tuberkulose. Im Jahre 2012 wurde er ständiger Vertreter des tionsgenetik und Infektiologie der Deutschen Gesellschaft Programmdirektors des Bereichs „Infektionen“ am FZB und für Hygiene und Mikrobiologie e.V. BIOANALYTICAL CHEMISTRY • DR. DOMINIK SCHWUDKE ROLE OF LIPIDS IN HOST-PATHOGEN INTERACTION STRUCTURE-FUNCTION RELATIONSHIP OF BACTERIAL PAMPS LIPIDOMICS NUCLEAR MAGNETIC RESONANCE SPECTROSCOPY MASS SPECTROMETRY MISSION THE DIVISION OF BIOANALYTICAL CHEMISTRY FOCUSES ON THE IDENTIFICATION AND QUANTITATION OF MEMBRANE COMPONENTS AND IMMUNOLOGICALLY ACTIVE BIOMOLECULES. WE ARE SPECIALIZED IN THE ANALYSIS OF LIPIDS, GLYCOCONJUGATES, AND CELL WALL COMPONENTS. WE AIM TO REVEAL SIGNALING CASCADES AND BIOSYNTHETIC PATHWAYS, WHICH ARE ALTERED DUE TO INFLAMMATION AND INFECTION. FOR THAT, WE DEVELOP AND APPLY OMICS WORKFLOWS AND METHODS FOR STRUCTURAL CHARACTERIZATION. MOST IMPORTANT FINDINGS gies against M. tuberculosis, we require a better understan- Lipid metabolic perturbations in epithelial tissue due to aller- ding how the pathogen interacts with the cell membranes and gic reaction can be identified with LC-MS receptors of the host. For that we advance analytical approa- Airway epithelial cells play an important role in the pathogene- ches using mass spectrometry (MS) and nuclear magnetic re- sis of inflammatory lung diseases such as asthma, cystic fibrosis sonance (NMR). We have developed a fast and reliable sample and COPD. In order to determine the lipidome in airway epithelia, preparation strategy for analysis of clinical isolates. With our we used an LC-MS based approach that was combined with a analytical strategy approximately 500 lipids can be identified. specific isolation procedure to obtain epithelial cells. In response We can show that the lipid profiles are sufficient to differen- to allergen challenge, perturbations in the sphingolipids were tiate strains using mycolic acids and sulfolipid profiles. Such detected, which led to increased levels of ceramides (Figure 1). approach opens now the possibility to link pathogenicity and resistance to the M. tuberculosis lipidome. Structural characterization of teichoic acids of Streptococcus pneumoniae Teichoic acids (TA) and peptidoglycan (PGN) are important structures to maintain cell integrity and cell morphology of Gram-positive bacteria. In cooperation with the group of Prof. Dr. S. Hammerschmidt (University of Greifswald) we investigate the structure and function of teichoic acids, as well as properties of cell wall modifying enzymes from the human lung pathogen S. pneumoniae. We refined the structural model of the pneumococcal LTA (Figure 2) and the elucidation of the so far unresolved chemical linkage of the WTA (which comprises Figure 1: Lipid class distribution in pmoles/sample determined in structurally identical repeating units) to the peptidoglycan is murine tracheal epithelial cells of animals challenged using house almost completed. The obtained results provide the basis for dust mite extract (red) and healthy controls (blue). The inset shows the a better understanding of the biosynthesis of TAs in S. pneu- single species distribution of ceramide species normalized to control moniae and led us already to the investigation of candidate samples (Zehethofer et al. 2014). proteins belonging to the LytR-Cps2A-Psr (LCP) protein family, We expanded the scope of this approach analyzing human bron- which are involved in connecting the WTA as well as the capsu- chus samples without pathological findings of adenocarcinoma le to the peptidoglycan. patients. For the human lung epithelium an unusual lipid class The pneumococcal PGN processing carboxypeptidases DacA distribution was found in which ceramide was the predominant and DacB are essential to maintain bacterial shape sphingolipid. To further our understanding of interaction of lung The inactivation of DacA (D,D-CPase), DacB (L,D-CPase; lipo- epithelial cells with exogenous stimuli we will combine laser cap- protein) or both enzymatic functions by gene knockouts led to ture microdissection with sensitive LC-MSn approaches. abnormal bacterial shape. The analysis of the PGN-peptides Structure-function analysis of mycobacterial cell wall components by high resolution mass spectrometry revealed new muropep- To develop diagnostic tools as well as new therapeutic strate- tides occurring in the dac-mutants, indicating that the compo- Seite 29 / PRA Infections BIRTE BUSKE • LARS FLORIAN EGGERS • DR. NICOLAS GISCH • HEIKO KÄSSNER • DR. MATTHIAS KRAJEWSKI • BRIGITTE KUNZ • CHAKRAVARTHY MARELLA • VERENA SCHOLZ • URSULA SCHOMBEL • SIMONE THOMSEN • FRANZISKA WALDOW • MICHAEL WEINKAUF • DR. NICOLE ZEHETHOFER pathophysiological effects of disabled cell wall hydrolases DacA and DacB. MOLECULAR MICROBIOLOGY 2014 Sep;93(6): 1183–1206. Figure 2: Structural model for pneumococcal LTA. Repeating units (RU) contain a pseudo-pentasaccharide (AATGalp – Glcp – Rib-ol-5-P – 6-O-P-Cho-GalpNAc – 6-O-P-Cho-GalpNAc), the terminal RU can occur with or without 6-O-P-Cho-substitution. Only the first repeating unit is β-1-linked to the lipid anchor (glucose-diacylglycerol), all other RUs are α-1-linked to the previous one. Hydroxyl groups of Rib-ol-5-P are substituted by D-alanine (D-Ala) potentially. The chain length of pneumococcal LTA is 4 to 8 RU (Gisch, Kohler et al., 2013). Zehethofer N, Bermbach S, Hagner S, Garn H, Müller J, Goldmann T, Lindner B, Schwudke D*, König P*. Lipid analysis of airway epithelial cells for studying respiratory diseases. CHROMATOGRAPHIA Nov 2014, doi:10.1007/s10337-014-27875) (* corresponding authors) sition of the elastic PGN macromolecule defines not only the architecture of the pneumococcal cell wall, but also the shape of the bacteria. Strikingly, the low surface abundance of nonlipidated DacB precursor protein in Lgt-deficient pneumococci was sufficient to maintain the wild-type cell shape and peptido- Hogendorf WFJ, Gisch N, Schwudke D, Heine H, Bols M, Pedersen CM. Total synthesis of five lipoteichoic acids of Clostridium difficile. CHEMISTRY – A EUROPEAN JOURNAL 2014 Oct;20(42):13511–13516. glycan architecture in this mutant, suggesting that the improper processing and anchoring of DacB has no major impact on INTERNAL AND EXTERNAL COLLABORATIONS DacB activity. This in turn indicates that lipidation of DacB is Inhouse Heine, Holst, Mamat, Reiling, Niemann, Gutsmann, Schromm, Grassl, Schaible, Jappe, Goldmann, GonzalezRoldan not necessary for its enzymatic activity (Abdullah et al., 2014). SELECTED PUBLICATIONS Gisch N*, Kohler T*, Ulmer AJ, Müthing J, Pribyl T, Fischer K., Lindner B, Hammerschmidt S, Zähringer U. Structural reevaluation of Streptococcus pneumoniae lipoteichoic acid and new insights into its immunostimulatory potency. JOURNAL OF BIOLOGICAL CHEMISTRY 2013 May;288(22):15654–15667. (* first authors) Hebbar S, Schulz WD, Sauer U, Schwudke D. Laser capture microdissection coupled with on-column extraction LC-MS(n) enables lipidomics of fluorescently labeled Drosophila neurons. ANALYTICAL CHEMISTRY 2014 Jun;86(11):5245–5252. Abdullah MR, Gutiérrez-Fernández J, Pribyl T, Gisch N, Saleh M, Rohde M, Petruschka L, Burchhardt G, Schwudke D, Hermoso JA, Hammerschmidt S. Structure of the pneumococcal L,D-carboxypeptidase DacB and National Hammerschmidt, Kohler (Greifswald), Hakenbeck, Denapaite (Kaiserslautern), Hölzl (Bonn), Ranf (München), Röder, Laudes (Kiel), König, Sadik, Ibrahim, Schwaninger (Lübeck), Haas (Bonn), Herker (Hamburg), Hebbar (Dresden) International Hermoso (Madrid), Pedersen (Kopenhagen), Vollmer (Newcastle), Baird (Bangor), Turska-Szewczuk, Komaniecka (Lublin) GRANT SUPPORT Exzellenzcluster Inflammation at Interfaces (CL X), SFB TR 22 (Z06; 2012-2013), SPP 1580 (2011-2014), DZIF MycoLip, DFG (GI 979/1-1; 2014-2017) BIOINFORMATICS • DR. KURT FELLENBERG DATA WAREHOUSING STATISTICS EXPLORATORY DATA ANALYSIS DATA VISUALIZATION MACHINE LEARNING DATA MINING HIGH-THROUGHPUT DATA MISSION IN MODERN LABORATORIES, SO-CALLED HIGH-THROUGHPUT TECHNIQUES SUCH AS MICROARRAY PLATFORMS OR NEXTGENERATION SEQUENCING RECORD THE STATUS OF THOUSANDS OF GENES AT ONCE. DATASETS THUS ACQUIRED ARE TOO LARGE FOR VISUAL INSPECTION. THEIR INTERPRETATION OFTEN BECOMES A BOTTLENECK. THE BIOINFORMATICS JUNIOR GROUP FOCUSES ON STORAGE, PROCESSING, AND INTERPRETATION OF HIGH-THROUGHPUT DATA. TO THIS END, WE APPLY METHODOLOGY FROM COMPUTER SCIENCE AND STATISTICS, AS WELL AS CONSIDERABLE COMPUTING POWER. MOST IMPORTANT FINDINGS visualizing variables together in one and the same plot, revealing Many of our projects could be summarized as extracting associations both among and between e.g. patients, treatments, patterns and coherences from quantitative data. In Borstel, and cytokines. However, not all statistically significant findings are several groups acquire microarray data (different platforms), also interesting. Interpretation requires close collaboration with contingency tables comprising cell frequencies, or other experts in the particular context under study, usually the people quantitative scores (e.g. by Luminex). Soon there will be RNA who acquired the data. Prior to any visualization and discussion, Sequencing of messenger as well as small regulatory RNA. however, the data need to be prepared (“preprocessed”) in order Coherences among and between at minimum two variables (e. to become comparable to each other at all. This is necessary g. genes × experiments) are extracted. Often, three variables because all of these data are subject to both noise and, what (e.g. patients × cell treatments × cytokines) have been cross- is worse, systematic errors. The sets of typical systematic errors tabulated. In other cases, interpretation is possible only by differ from platform to platform. One well-known example of a accounting for global players “behind” the objects under study. systematic error is the label-incorporation rate for all microarray Those can be clinical parameters characteristic for a group of platforms. Diverging labeling efficiencies cause multiplicative patients, or Gene Ontology terms common and discriminative offsets between the resulting signals. In other words, numbers for a cluster of genes exhibiting a particular expression profile recorded by individual microarrays never have the same (Figure 1). As a means of exploratory data analysis, we aim at scale. Thus, each individual dataset needs to be rigorously preprocessed before anything can be learned from it by machine learning or any statistical test. At times, (pre-)processing data can be a sufficient goal in and of itself, ending up in a user-friendly software. Next-generation sequencing with its unprecedented level of detail promises to become an important tool for tuberculosis lineage and antibiotic resistance diagnosis. However, while having access to the technique often does not present an insurmountable obstacle any more, data analysis is still daunting to many of its users. One sample yields millions of short sequences (called “reads”) that are prone to sequencing errors. Here, typical systematic errors can occur while aligning the reads to to a reference genome, in close vicinity to insertions or deletions. In order not to wrongly postulate a variant (type I error) and thus in some places resistance to an antibiotic, the alignment needs to be corrected at these positions. Another Figure 1: Microarray experiments (colored spheres, budding yeast transfected with CDC14 under the control of a galactose inducible potential source of false positive results is that base qualities are generally over-estimated by sequencers and thus have to promoter), genes (white dots) and gene ontology terms (light blue) be re-calibrated to become a realistic basis for the subsequent projected in one and the same 3D plot. variant detection. However, it is equally inconvenient (if not Seite 31 / PRA Infections VIOLA SCHLEUSENER worse) to overlook a present antibiotic resistance (type II error). A prerequisite for successful next generation sequencing Our software warns the user whenever this is possible, such as workflows is to apply sufficient underlying computational when genomic positions of resistance mediating genes are not resources. We started to build up an IT infrastructure based sufficiently covered by sequencing reads. on two IBM Servers with in total 56 physical (i.e. before hyperthreading) CPU cores and 600 Gb memory. Sometimes, a particular calculation requires considerable CPU time or a large chunk of memory. In order to grant the flexibility to apply these resources where needed by us and other groups, dedicated virtual servers such as microarray, ftp, web, and database server have been installed (9 in total). Available services include web interfaces e.g. for local blast, gene wide association studies, or abovementioned antibiotic lineage and resistance determination. There are also client/server analysis systems (ideal for collaboration analyses of distributed groups, inside and outside FZB), for microbial outbreak investigation (Ridom Seqsphere) and for microarray data analysis (http:// mchips.org). Apart from flexibility and optimal use of resources Figure 2: A web interface (http://phyresse.org) computes Mycobacterium tuberculosis genotype and antibiotic resistance from uploaded next generation sequencing data. Evaluation has confirmed that obtained results are superior by load-balancing, server virtualization also provides a fallback solution for each service in case one of the two physical servers may fail. Storage space is supplied by to date two pairs of mirrored file servers, by and large 100 Tb. to both classical Sanger sequencing and conventional typing methods, e.g. by detection of heterogenous variants and SELECTED PUBLICATIONS improved phylogenetic lineage classification. Providing state-of- Schröder C, Srinivasan H, Sill M, Linseisen J, Fellenberg K, Becker N, Nieters A, Hoheisel JD. Plasma protein analysis of patients with different B-cell lymphomas using high-content antibody microarrays. Proteomics Clin Appl. 2013 Dec;7(11-12):802-12. the-art methodology to life scientists and physicians untrained in bioinformatics, the analysis pipeline is run automatically. Making it available as a web service, we furthermore relieve its users from any local installation and systems administration. Being accessible via the Internet, a running network connection remains the only requirement. After selecting the files to upload, one mouse click invokes the entire workflow with no further action being required from the user. Afterwards, results are available as plain language (HTML) reports in which we invested a considerable share of developing time to layout in a clear, Dunkel N, Biswas K, Hiller E, Fellenberg K, Satheesh SV, Rupp S, Morschhäuser J. Control of morphogenesis, protease secretion and gene expression in Candida albicans by the preferred nitrogen source ammonium. Microbiology. 2014 Aug;160(Pt 8):1599-608. self-explanatory and simple way. The Phylo-Resistance Search Engine is accessible free of charge and without registration at INTERNAL AND EXTERNAL COLLABORATIONS http://phyresse.org. Uploaded data and computed results are S Niemann, N Reiling, K Brandenburg, T Gutsmann, A Schromm, C Lange, C Herzmann, J Behrends, B Schneider, H Heine, T Goldmann, M Reinhold carefully isolated by secure sessions in order to prevent access by other users (Figure 2, red-framed area). Motivated by the fact that resistant bacteria may represent only a small share of the sample prior to antibiotic treatment, we currently investigate how to discriminate the resulting low-frequency variants from mere sequencing errors. S Rupp, Fraunhofer IGB, Stuttgart; C Köser, University of Cambridge, UK; C Schröder, Sciomics GmbH, Heidelberg; J Hoheisel, DKFZ, Heidelberg BIOPHYSICS • PROF. DR. THOMAS GUTSMANN LIPID MEMBRANES HOST DEFENSE PEPTIDES SIGNAL TRANSDUCTION GLYCOLIPIDS LIPID-PEPTIDE INTERACTIONS MISSION THE FUNCTION OF LIPIDS AND LIPID MEMBRANES AND THEIR INTERACTION WITH PEPTIDES AND PROTEINS IN THE CONTEXT OF LUNG INFECTIONS IS IN OUR FOCUS. TO ANSWER BIOMEDICAL QUESTIONS AND TO DEVELOP NEW THERAPEUTICS WE FIRST AIM AT AN UNDERSTANDING OF THE UNDERLYING MOLECULAR FUNCTION OF MEMBRANES AND PEPTIDES. THE ORIGIN AND THE BIOMEDICAL CONTEXT OF THE PEPTIDES CAN BE VERY DIFFERENT; HOWEVER, WE ARE INTERESTED IN THE COMMON STRUCTURE-FUNCTION RELATIONSHIP. WE USE A NUMBER OF BIOLOGICAL AND BIOPHYSICAL TECHNIQUES TO CHARACTERIZE BACTERIAL AND HUMAN IMMUNE CELL MEMBRANES AND IN PARTICULAR RECONSTITUTED MEMBRANES MIMICKING THEIR NATURAL EXAMPLES. THE BIOMEDICAL FOCUS IS DIRECTED TOWARDS: (A) THE ORGANIZATION AND PROPERTIES OF MEMBRANES COMPOSED OF PHOSPHO- AND GLYCOLIPIDS; (B) THE FUNCTION OF PORE-FORMING PEPTIDES AND PROTEINS, E.G. HOST DEFENSE PEPTIDES (HDPS) OF THE INNATE IMMUNE SYSTEM IN THE LUNG; (C) FURTHER ACTIVITIES OF MEMBRANE ACTIVE PEPTIDES, E.G. NEUTRALIZATION OF LIPIDIC MOLECULES SUCH AS ENDOTOXINS AND LIPOPROTEINS AND INVOLVEMENT IN MEMBRANE-ASSOCIATED SIGNAL TRANSDUCTION PATHWAYS.. systems the importance of the defensin-typical three disulfide MOST IMPORTANT FINDINGS bonds for the activity of the AMPs was investigated. The study Pore-forming peptides and proteins play an important role in demonstrates that the refining HDP structures can generate the battle between the human immune system and microbes. more active compounds against certain specific strains. The HDPs can kill extra- and intracellular microbes by forming po- binding strength between the peptides and LPS membranes res in the microbial cell envelope. Furthermore, bacterial po- is decisive for the permeabilization of the outer membrane, rins are important for the uptake of classical antibiotics. On but for the antimicrobial activity further peptide parameters, the other hand, also microbes can kill other microbes by pore- e.g. peptide flexibility, are important. formers. In the last years, we investigated pore-forming prote- Pore Formation by the Virulence-assocoiated protein A (in ins and peptides of microbial origin, which are supposed to collaboration with A. Haas) be important for the intracellular survival of lung pathogens. Pore-forming activities are on the one hand important mechanisms to kill microbes by host defense peptides but on the other hand, microbes seem to use pore-forming proteins and peptides to avoid the acidification of the phagosome. The major aim of this project is to characterize the pore-forming mechanisms and how microbial and phagosome membranes can resist the activity of their own pore-formers. Together with our collaboration partner Albert Haas (University of Bonn) we characterized the membrane activity of VapA from Rodococcus equi. Using experiments on reconstituted model membranes for the phagosome membrane we analyzed the pore forming activity and also the general mode of action of the virulence-associated protein A (VapA) which is expressed in Figure 1: Summary of various pore-forming processes in the interaction between bacteria and host cells. the membrane of R. equi and released into the phagosome. Atomic force microscopic analysis of these membranes Structure-Function Relationship between Human Beta Defen- revealed a strong aggregation effect of VapA to certain lipid sin-3 and Membranes domains. This induced a reorganization of the domain struc- One of the most efficient human HDP is the human beta de- ture of cholesterol-containing bilayers. Different permeability fensin-3 (hBD-3) which is produced by, e.g. keratinocytes and increasing effects induced by VapA in reconstituted phagoso- lung epithelial cells. In biological assays as well as in biophy- me membrane were observed with electrophysiological mea- sical experiments utilizing different membrane reconstitution surements on freestanding membranes. Based on these fin- Seite 33 / PRA Infections PROF. DR. KLAUS BRANDENBURG • SABINE DABELSTEIN • NADINE GEBAUER • SABRINA GROTH • CHRISTINE HAMANN • DR. LENA HEINBOCKEL • LAURA PAULOWSKI • DR. ANNIKA SAATHOFF • CHRISTIAN NEHLS • NIKLAS STEIN • KERSTIN STEPHAN dings we suggest a five-step model for the interaction between spectrum neutralizing activity of peptide Pep19-2.5 on bac- VapA and the phagosome membrane (binding to membrane terial pathogenicity factors. Antimicrob Agents Chemother. – weak intercalation – aggregation – membrane permeabi- 2013, 57(3):1480-1487. lization – membrane destruction). The results indicate that the interaction between VapA and the phagosome membrane Heinbockel L, Palacios-Chaves L, Alexander C, Rietschel E, works contrary to several essential increments of phagosome Behrends J, Correa W, Fukuoka S, Gutsmann T, Ulmer AJ, maturation. Brandenburg K. Mechanism of Hbγ-35-induced an increase in the activation of the human immune system by endotoxins. Innate Immun. 2014 Jul 16. INTERNAL AND EXTERNAL COLLABORATIONS Internal cooperation: Norbert Reiling, Ulrich Schaible, Niels Röckendorf and Andreas Frey, Otto Holst, Torsten Goldmann and Ekkehard Vollmer Andra Schromm External cooperation: R. Schmitz-Streit, University of Kiel; T. Lindhorst, University of Kiel, M. Leippe; University of Kiel; J. Grötzinger, University of Kiel; C. Hübner, University of Lübeck; M. Hornef, Medical School Hannover; T. Schürholz, Medical School Aachen; A. Haas, University of Bonn; B. Hube, Hans Knöll Institut Jena; M. Winterhalter, Jacobs University, Bremen; G. Martinez de Tejada, Universidad de Navarra, Pamplona Figure 2: Membrane activity of VapA. The two atomic force microscopic images show a cholesterol-rich solid supported bilayer (DOPC:SM:Chol 9:9:2) on the left side and the same membrane after addition of VapA on the right side, demonstrating an aggregation of the small proteins GRANT SUPPORT Deutsche Forschungsgemeinschaft (GU 568/4-1, ‚Biophysical investigations into the interactions between antimicrobial pep- and a modification of the cholesterol-rich domains. The electrophysiolo- tides of the epithelial defense and microbial cell envelopes’; gical data demonstrate the pore-forming activity of VapA. Schwerpunktprogramm 1580 „Intracellular compartments as places of pathogen-host-interactions“) SELECTED PUBLICATIONS Leibniz-Graduiertenschule (Adhesion of Plasmodium-infected Michalek M, Sönnichsen FD, Wechselberger R, Dingley AJ, erythrocytes to the human endothelial receptors CD36, and Hung CW, Kopp A, Wienk H, Simanski M, Herbst R, Loren- P-selectin - Characterization of binding properties) zen I, Marciano-Cabral F, Gelhaus C, Gutsmann T, Tholey A, Else Kröner-Fresenius-Stiftung (Biophysikalische Charakteri- Grötzinger J, Leippe M. Structure and function of a unique sierung der Interaktion von Endotoxinen mit nicht-steroiden pore-forming protein from a pathogenic acanthamoeba. Nat antientzündlichen Wirkstoffen und deren Modifikation durch Chem Biol. 2013, 9(1):37-42. antimikrobielle Peptide) Cluster of Excellence „Inflammation at interfaces“ (Anti-in- Heinbockel L, Sánchez-Gómez S, Martinez de Tejada G, flammatory regulation of immune cells by membrane active Dömming S, Brandenburg J, Kaconis Y, Hornef M, Dupont host defense peptides) A, Marwitz S, Goldmann T, Ernst M, Gutsmann T, Schürholz T, Brandenburg K. Preclinical investigations reveal the broad- CELLULAR MICROBIOLOGY • PROF. DR. ULRICH E. SCHAIBLE MYCOBACTERIUM TUBERCULOSIS INTRACELLULAR NICHE PHAGOSOME MACROPHAGE NEUTROPHIL MICROBIOTA LIPIDS MISSION THE GROUP CELLULAR MICROBIOLOGY STUDIES HOST-PATHOGEN INTERACTIONS IN TUBERCULOSIS ON THE MOLECULAR, CELLULAR AND ANIMAL MODEL LEVEL BY FOCUSING ON THE QUESTION HOW THE INTRACELLULAR NICHE OF THE PATHOGEN INFLUENCES MYCOBACTERIAL SURVIVAL, INNATE AND ACQUIRED IMMUNITY, PATHOGENESIS AND DRUG EFFICACY. MOST IMPORTANT FINDINGS et al., 2013). Phagosome maturation is a hallmark for anti- The causative agent of tuberculosis (TB), Mycobacterium (M.) mycobacterial macrophage function via sequestration of iron, tuberculosis, is a facultative intracellular pathogen of resting a growth factor for mycobacteria, and exposure to lysosomal macrophages. Blocking phagosome maturation allows intra- enzymes. As free iron is a driver of bacterial growth, we deve- cellular growth of mycobacteria and affects host responses. loped, together with colleagues at Kings College London, novel In contrast, IFN-γ activated macrophages are equipped with iron sensors to measure free iron in patient samples (Ma et al., effectors to counteract bacterial virulence and promote pha- 2014). Regarding lysosomal enzymes in host responses to my- gosome maturation (Weiss & Schaible, 2015). While macro- cobacteria, we identified the lysosomal phospholipase A2 as phage activation by Th1 responses are well established for important for proper Th1 responses to M. tuberculosis (Schnei- protective immunity in TB, and the prime immune target of anti-TB vaccination, the role of antibodies and neutrophils in M. tuberculosis control is less clear. Based on our finding that the mycobacterial cell wall glycolipid, trehalose dimycolate (TDM), blocks phagosome maturation, we observed that TDM also interferes with phagosome biogenesis upon antibody-Fc receptor-mediated phagocytosis. This virulence function of TDM requires its receptor, Mincle, the signaling molecule SHIP1 as well as the inhibitory FcγRIIB (Patin et al., submitted). Interference of TDM with antibody-FcR mediated elimination of M. tuberculosis indicates that specific antibodies add little to protective immunity in TB. Using proteomics of isolated phagosomes we identified the M. tuberculosis-host cell interactome including host cell cytoskeleton elements such as actin and the WASH complex to be associated with mycobacterial phagosomes. Blocking actin polymerization promoted mycobacterial phago-lysosome fusion and killing of mycobacteria (Kolonko Figure 2: a. FISH analysis of a lung section from a C57BL/6 mouse raised under environmental conditions using a global eubacterial 16S rRNA probe depicting lung microbiota (red) on alveolar epithelia (blue Figure 1: Imaging efferocytosis: 3D reconstruction of an apoptotic human neutrophil (green) infected with M. bovis BCG (red), which has been efferocytosed by a human macrophage (blue: NADPase oxidase). = DAPI stain of nuclei). b. Distribution of bacterial phyla within the murine lung microbiota based on 16S rRNA 454 sequencing. Seite 35 / PRA Infections DR. CHRISTIAN ALEXANDER • JANNIKE BLANK (04/14) • MAIKE BURMEISTER • DR. TOBIAS DALLENGA • JACQUELINE EICH • ANNA CHRISTINA GEFFKEN • NINA GROHMANN • KRISTINE HAGENS • NADINE HARMEL (04/14) • DAGMAR MEYER • CARLOTTA OBER-BLÖBAUM • BHESH PAUDYAL • NATALJA REDINGER • STEFFI RENK ((09/14) • DR. BIANCA SCHNEIDER (04/14) • DR. YEOJUN YUN • DR. NICOLE ZEHETHOFER der et al., 2014). Neutrophils, the main infected cell population Schneider BE, Behrends J, Hagens K, Harmel N, Shayman in active TB patients, quickly succumb to necrotic cell death JA, Schaible UE. 2014 Lysosomal phospholipase A2: A novel upon infection with virulent but not attenuated M. tuberculosis player in host immunity to Mycobacterium tuberculosis. Eur J strains (Corleis et al. 2012). Thus, neutrophils contribute to Immunol. 44(8): 2394 exacerbated pathology rather than protective immunity. Removal of neutrophils by efferocytosis promotes mycobacterial Weiss G, Schaible UE. 2015 Macrophage defense mechanisms survival in macrophages (Dallenga et al., submitted; Figure 1). against intracellular bacteria Immunological Reviews in press The rise of multi-drug resistant M. tuberculosis strains requires intensified search for novel vaccination and treatment strate- Yun Y, Srinivas G, Kuenzel S, Linnenbrink M, Alnahas S, Bru- gies. Within national and international networks (CNV, DZIF- ce KD, Steinhoff U, Baines JF, Schaible UE. 2014 Environ- TTU-TB, NAREB), we use M. tuberculosis reporter strains and mentally determined differences in the murine lung microbio- in vivo imaging (Andreu et al., 2013) to study novel nanopartic- ta and their relation to alveolar architecture. PlosOne in press le-based vaccine and drug formulations. We hypothesize that nanoparticles improve treatment by targeting the intracellular INTERNAL AND EXTERNAL COLLABORATIONS niche of M. tuberculosis. The respiratory tract microecology is Internal collaborations a neglected determinant for TB pathogenesis. Using mice from Guntram Graßl, Thomas Gutsmann, Christoph Hölscher, Uwe different environments, we pivotally proved the presence of mi- Mamat, Stefan Niemann, Otto Holst, Norbert Reiling, Bianca crobiota in the mammalian lung by culture, 16S rRNA sequen- Schneider, Dominik Schwudke, cing and FISH (Yun et al., 2014; Figure 2). The identification of External collaborations a lung microbiota opens new perspectives for studies on its •Iron: Robert Hider, London role in lung function and infectious and non-infectious inflam- •Microbiota: John Baines, Plön; Kenneth Bruce, London; mation (including TB and asthma) and immunity. Ulrich Steinhoff, Marburg, Frank Schmidt, Greifswald; Wolfgang Streit, Hamburg; Jörg Steinmann, Essen SELECTED PUBLICATIONS •Lysosomes: James Shayman, AnnArbor; Dieter Adam, Kiel Andreu N, Zelmer A, Sampson SL, Ikeh M, Bancroft GJ, •Pathogen-Host Interactome: Dörthe Becher, Greifswald; Mo- Schaible UE, Wiles S, Robertson BD. 2013 Rapid in vivo assessment of drug efficacy against Mycobacterium tubercu- nika Hagedorn, Hamburg; Albert Haas, Bonn •Nanoparticles: Lea Ann Dailey, London losis using an improved firefly luciferase. J Antimicrob Chemother. 68(9): 2118 GRANT SUPPORT BMBF: Medical Infection Pathogenomics (M. tuberculosis- Corleis B, Korbel D, Wilson R, Bylund J, Chee R, Schaible Host Interactomes); German Center of Infection (DZIF), TTU-TB UE. 2012 Escape of Mycobacterium tuberculosis from oxidati- (MycoLip) ve killing by neutrophils. Cell Microbiol 14(7): 1109 DFG: Specific Priority Program 1580 Intracellular compartments as places of pathogen-host-interactions; Kolonko M, Geffken AC, Blumer T, Hagens K, Schaible UE, Project grant (Role of lysosomal phospholipases in infection); Hagedorn M. 2013 WASH-driven actin polymerization is requi- Excellence Cluster Inflammation at Interfaces (Lysosomal red for efficient mycobacterial phagosome maturation arrest. disorders and bacteria-induced inflammation); IRTG 1911 Cell Microbiol. 16(2): 232 Immunoregulation of Inflammation in Allergy and Infection; Danish Research Council: Center of Nano-vaccines (Imaging Ma Y, Podinovskaia M, Evans PJ, Emma G, Schaible UE, vaccine efficacy) Porter J, Hider RC. 2014 A novel method for non-transferrin- EU FP7: NAREB (Nanotherapeutics to treat bacterial bound iron quantification by chelatable fluorescent beads infectious diseases) based on flow cytometry. Biochem J. 463(3): 351 CELLULAR PNEUMOLOGY • PROF. DR. CORDULA STAMME INFECTION SURFACTANT PROTEINS ALVEOLAR MACROPHAGES INNATE IMMUNE MEMBRANE TRAFFICKING MISSION DEFENSE STRATEGIES THAT BOOST ENDOGENOUS ELEMENTS OF INNATE LUNG RESPONSES TO INFECTION ARE A MAJOR FOCUS (BOTH BASIC AND TRANSLATIONAL) TO DEVELOP NEW PROPHYLACTIC OR THERAPEUTIC MEASURES. THE HUMAN C-TYPE LECTINS SURFACTANT PROTEIN (SP)-A AND SP-D ARE SOLUBLE PATTERN RECOGNITION RECEPTORS REGULATING LUNG IMMUNE RESPONSES IN VIVO, PARTIALLY BY SPECIFICALLY CONTROLLING THE HIGH PLASTICITY OF ALVEOLAR MACROPHAGES TOWARDS LUNG IMMUNE HOMEOSTASIS. WE INVESTIGATE LUNG SPECIFIC INNATE IMPRINTING BY SP-A AND SP-D TO PULMONARY INFECTION WITH THE FOCUS ON LUNG CELL-SPECIFIC/LECTIN INTERACTIONS AND DOWNSTREAM EFFECTS ON CELL ACTIVATION/DEACTIVATION TO TARGET FOR DEVELOPMENT OF THERAPEUTIC INTERVENTIONS. MOST IMPORTANT FINDINGS Collectin-directed receptor trafficking Lung infection by Gram-negative bacteria is a major cause of morbidity and mortality in humans. Lipopolysaccharide (LPS), located in the outer membrane of the Gram-negative bacterial cell wall, is a highly potent stimulus of immune and structural cells via the TLR4/MD2 complex whose function is sequentially regulated by defined subsets of adaptor proteins. In general, the role of lung-specific microenvironmental factors in regulating expression and intracellular TLR4 localization in response to LPS is only beginning to be experimentally addressed. The so far best-studied microenvironmental factor modulating TLR4 signaling in the lung is pulmonary surfactant. Pulmonary SP-A modulates the alveolar macrophage Figure 1: LPS-induced TNFα release in BALF from β-arrestin 2-/- mice is accelerated and more pronounced than in WT mice. threshold of LPS activity both in vitro and in vivo through va- gnaling of LPS-induced TLR4 in vitro and in vivo engaging rious mechanisms. Data from our group demonstrated that β-arrestin 2. SP-A modulates the spatiotemporal compartmentalization of LPS-induced TLR4 in primary alveolar macrophages in vitro Alternative functions of endocytic proteins and in a mouse model of intratracheal LPS challenge SP-A Lung immune host defense functions of SP-A are partly re- reduces the LPS-induced co-localization of TLR4 with early gulated by clathrin-mediated endocytosis (CME)-dependent endosomes and promotes TLR4 localization with the Golgi uptake of the protein by alveolar macrophages (Fig.2). The thereby inhibiting TLR4 signaling. Furthermore, the lack of central CME proteins clathrin and the adaptor protein com- direct SP-A/TLR4 co-localization, the SP-A-mediated upregu- plex AP2, however, have secondary roles in cellular homeo- lation of β-arrestin 2, a pivotal negative regulator of TLR4- stasis that are independent of their endocytic functions. We dependent responses, and the SP-A-enhanced β-arrestin2/ dissected endocytic from non-endocytic functions of CME TLR4 interaction suggest that SP-A-modulated cellular distri- proteins in constitutive and SP-A-modulated LPS signaling. bution of TLR4 in primary alveolar macrophages is mediated Demonstrated by pharmacological inhibition and RNAi in indirectly by integrating β-arrestin 2 scaffolding interactions. primary AM and RAW264.7 cells, respectively, clathrin hea- Pulmonary LPS-induced TNFα release in β-arrestin 2 mice is vy chain (CHC) and the α-adaptin subunit of the AP2 com- accelerated and enhanced (Fig. 1) and exogenous SP-A fails plex, but not dynamin, prevent basal IκB-α degradation and to inhibit both lung LPS-induced TNF-α release and TLR4/ TNFα release independent of their canonical role in mem- endosome positioning. Our data demonstrate that SP-A mo- brane trafficking. SP-A post-trancriptionally and transiently dulates the spatiotemporal compartmentalization and si- enhances the basal protein expression of CHC in primary AM -/- Seite 37 / PRA Infections DOMINIKA BIEDZIAK • KATJA FREUNDT • KARINA GREVE • DR. PATRICK KELLNER Sender V, Stamme C. Lung cell-specific modulation of LPSinduced TLR4 receptor and adaptor localization. Commun Integr Biol. 2014 May 16;7:e29053. doi: 10.4161/cib.29053. INTERNAL AND EXTERNAL COLLABORATIONS Internal: PD Dr. N. Reiling, Microbial Interface Biology, Dr. C. Steinhäuser, Microbial Interface Biology, Prof. Dr. O. Holst, Structural Biochemistry. National: Prof. Dr. C. Hübner, Dept. of Physics, Univ. Lübeck, Prof. Dr. C. Nau, Dept of Anaesthesiology, Univ. Lübeck, Prof. Dr. U. Costabel/Dr. F. Bonella, Ruhrlandklinik, Univ. Essen, Dr. Srikanth Karnati, Univ. Giessen. Figure 2: Dynasore inhibits SP-A uptake by primary alveolar macrophages. International: Prof. C. Casals and C. Minutti, Universidad Complutense de Madrid, Spain, Dr. V. Sender, Karolinska Institutet, Stockholm, Sweden, Prof. Dr. B. Beck-Schimmer, Univ from rats, SP-A+/+, SP-A , and β-arrestin 2 mice. Further-/- -/- Zürich, Switzerland. more, intratracheal application of exogenous SP-A enhances CHC expression in vivo. In contrast, LPS counteracts the GRANT SUPPORT effect of SP-A independent of animal species and mouse Work in this group was supported by the Deutsche For- genotype. The pharmacological inhibition of SP-A-specific si- schungsgemeinschaft (Grant 609/1-4 and 609/2-1 to C S; gnaling pathways abolishes SP-A-mediated increase in CHC. Grant 1999/2-1 to C M). Constitutive anti-inflammatory immunomodulation by SP-A depends on endocytic proteins whereas neither dynamin nor α-adaptin are required for SP-A-mediated inhibition of LPS-induced NF-κB activity and TNFα release. In summary, our data demonstrate that CHC and α-adaptin constitutively downregulate IκB-α/NF-κB activity in macrophages independent of their endocytic functions. In addition they provide first evidence that SP-A enhances the constitutive non-endocytic capacity of these proteins. Further characterization of the relevance of clathrin-mediated pathways in alveolar (patho)physiology not only will lead to better understanding of lung disease mechanisms but also might reveal specific clathrin and/or adaptor binding interactions to target for therapeutic strategies. SELECTED PUBLICATIONS Sender V, Lang L, Stamme C. Surfactant protein-A modulates LPS-induced TLR4 localization and signaling via β-arrestin 2. PLoS ONE 2013, 8(3):e59896. CLINICAL INFECTIOUS DISEASES • PROF. DR. MED. DR. H.C. CHRISTOPH LANGE DZIF MDR-TB INTERNATIONAL HEALTH & INFECTIOUS DISEASES IGRA NTM TUBERCULOSIS XDR-TB TBNET MISSION WE AIM TO IMPROVE THE PREVENTION, DIAGNOSIS AND TREATMENT OF TUBERCULOSIS (ESPECIALLY M/XDR-TB) AND RESPIRATORY DISEASES CAUSED BY NON-TUBERCULOUS MYCOBACTERIA AND TO INTEGRATE SCIENTIFIC ADVANCES INTO CLINICAL PRACTICE. MOST IMPORTANT FINDINGS clinical evidence. We summarised the current knowledge on Consensus statement to optimize the management for pati- the prevention, diagnosis and treatment of adults and child- ents with M/XDR-TB in Europe ren with M/XDR-TB and their contacts, and reached expert The emergence of multidrug-resistant tuberculosis (MDR-TB) consensus recommendations on questions where scientific and extensively drug-resistant (XDR)-TB substantially challen- evidence is still lacking. ges TB control, especially in the European Region of the World High percentages of primary bacillary transmission and ad- Health Organization, where the highest prevalence of M/XDR- vanced resistance to 2nd line anti-TB drugs characterize ca- TB cases is reported. The current management of patients with ses of MDR-TB in Europe M/XDR-TB is extremely complex for medical, social and public We evaluated risk factors and levels of second-line drug-re- health systems. The treatment with currently available anti-TB sistance in M. tuberculosis in a European cohort of patients therapies to achieve relapse-free cure is long and undermined with MDR-TB. 380 patients with MDR-TB and 376 patients by a high frequency of adverse drug events, suboptimal treat- with non-MDR-TB were enrolled at 23 centres in 16 European ment adherence, high costs and low treatment success rates. countries between 2010 and 2011. 52.4 % of MDR-TB pati- Availability of optimal management for patients with M/XDR- ents had never been treated for TB in the past, suggesting TB is limited even in the European Region. In the absence of a primary transmission of MDR M. tuberculosis. At initiation of preventive vaccine, more effective diagnostic tools and novel MDR-TB treatment 59.7 % of tested M. tuberculosis strains therapeutic interventions the control of M/XDR-TB will be very were resistant to pyrazinamide, while 51.1 % were resistant difficult. Despite recent scientific advances in M/XDR-TB care, to at least one 2nd line drug. 26.6 % showed resistance to 2nd decisions for the management of patients with M/XDR-TB and line injectables and 17.6 % to fluoroquinolones, 6.8 % were their contacts often rely on expert opinions, rather than on XDR-TB. Previous TB treatment (p<0.001) was the strongest risk factor for MDR-TB. In conclusion, high percentages of primary transmission and advanced resistance to 2nd line anti-TB drugs characterize cases of MDR-TB in Europe. The tuberculin skin test and interferon-γ release assays poorly predict the development of tuberculosis in immunocompromized hosts In the absence of active tuberculosis, a positive result in the tuberculin skin test (TST) or interferon-γ release assays (IGRAs) defines latent infection with Mycobacterium tuberculosis, although test results may vary depending on immunodeficiency. We compared the performance of TST and IGRAs in five different groups of immunocompromised patients, and evaluated their ability to identify those at risk for development of tuberculosis. Immunocompromised patients with HIV infection, chro- Figure 1: Patient care (lower right) and clinical training (lower left) at the Medical Clinic in Borstel and at international partner sites (UNAM in Windhoek, Namibia – upper left – and USMF in Chisinau, Moldova nic renal failure, rheumatoid arthritis, solid-organ or stem-cell transplantation, and healthy control subjects were evaluated -upper right) are core activities of the members of the Clinical head-to-head by the TST, QuantiFERON-TB-Gold in-tube test Infectious Diseases research group. (ELISA), and T-SPOT.TB test (enzyme-linked immunospot) at 17 Seite 39 / PRA Infections LUKAS BUSENBENDER • FRANZISKA DADUNA • STEFANIE DECKERT • CORDULA EHLERS • EMILIA HÄUSSINGER • DR. MED. CHRISTIAN HERZMANN • DR. MED. JAN HEYCKENDORF • JESSICA HOFMEISTER • DR. MED. ULF GREINERT • DR. MED. GUNAR GÜNTHER (MPH) • CHRISTIAN GUTSFELD • DR. MED. CLAUDIA JAFARI • DR. MED. BARBARA KALSDORF • DR MED. (BUCHAREST) IOANA OLARU • JULIANE RADLOFF • PROF. DR. MED. HANS RIEDER • DR. MED. HELMUT SALZER (MPH) • NELLEKE SMITSMAN • DR. MED. ELENA TERHALLE • DR. MED. NASSTASJA WASSILEW individuals and was poorly predicted by TST or IGRAs. SELECTED PUBLICATIONS Lange C, Abubakar I, Alffenaar JWC, Bothamley G, Caminero JA, Carvalho ACA, Chang KC, Luigi Codecasa L, Correia A, Crudu V, Davies P, Dedicoat M, Drobniewski F, Duarte R, Ehlers C, Erkens C, Goletti D, Günther G, Elmira Ibraim E, Kampmann B, Kuksa L, de Lange W, van Leth F, van Lunzen J, Matteelli A, Menzies D, Monedero I, Richter, Rüsch-Gerdes S, Andreas Sandgren A, Scardigli A, Skrahina A, Tortoli E, Volchenkov G, Wagner D, van der Werf MJ, Williams B, Yew WW, Zellweger JP and Cirillo DM for the TBNET. Management of patients Figure 2: Hypothetical model of biosignatures to individualize the duration of antituberculosis with M/XDR-TB in Europe. A TBNET therapy (Heyckendorf J. et al. Getting personal perspectives on individualized treatment duration in consensus statement. Eur Respir J multidrug-resistant and extensively drug-resistant tuberculosis. Am J Respir Crit Care Med 2014; 2014; Jul;44(1):23-63. 190(4):374-83). centers in 11 European countries. Development of tuberculo- Günther G, van Leth F, Alexandru S, Altet A, Avsar K, Bang sis was assessed during follow-up. Frequencies of positive test D, Barbuta R, Bothamley G, Ciobanu A, Crudu V, Davilovits M, results varied from 8.7 to 15.9 % in HIV infection (n = 768), Dedicoat M, Duarte R, Gualano G, Kunst H, de Lange W, Lei- 25.3 to 30.6% in chronic renal failure (n = 270), 25.0 % to mane V, Magis-Escurra C, McLaughlin AM, Muylle I, Polcová 37.2 % in rheumatoid arthritis (n = 199), 9.0 to 20.0 % in solid- V, Pontali E, Popa C, Rumetshofer R, Skrahina A, Solodov- organ transplant recipients (n = 197), 0 % to 5.8% in stem-cell nikova V, Spinu V, Tiberi S, Viiklepp P and Lange C, for the transplant recipients (n = 103), and 11.2 to 15.2 % in immuno- TBNET. Multidrug-resistant tuberculosis in Europe: A TBNET competent control subjects (n = 211). Eleven patients (10 with study. Emerg Infec Dis 2015 Mar;21(3):409-16 HIV infection and one solid-organ transplant recipient) developed tuberculosis during a median follow-up of 1.8 (interquarti- Sester M, van Leth F, Bruchfeld J, Bumbacea D, Cirillo DM, le range, 0.2–3.0) years. Six of the 11 patients had a negative Dilektasli AG, Domínguez J, Duarte R, Ernst M, Eyuboglu FO, or indeterminate test result in all three tests at the time of Gerogianni I, Girardi E, Goletti D, Janssens JP, Julander I, Lan- screening. Tuberculosis incidence was generally low, but hig- ge B, Latorre I, Losi M, Markova R, Matteelli A, Milburn H, her in HIV-infected individuals with a positive TST (3.25 cases Ravn P, Scholman T, Soccal PM, Straub M, Wagner D, Wolf T, per 100 person-years) than with a positive ELISA (1.31 cases Yalcin A and Lange C for the TBNET. Risk assessment of tu- per 100 person-years) or enzyme-linked immunospot result berculosis in immunocompromised patients. A TBNET study. (1.78 cases per 100 person-years). No cases of tuberculosis Am J Respir Crit Care Med 2014; Nov 15; 190(10):1168-76 occurred in patients who received preventive chemotherapy. Among immunocompromised patients evaluated in this study, GRANT SUPPORT progression toward tuberculosis was highest in HIV infected BMBF, DAAD, DZIF, EDCTP, EU-FP7, S.-H. TB Society, TBNET COINFECTION • DR. BIANCA SCHNEIDER TUBERCULOSIS MALARIA COINFECTIONS ANIMAL MODELS IMMUNE MODULATION HOST-PATHOGEN INTERACTION MISSION IN SOME AREAS OF THE WORLD, COINFECTIONS WITH TWO OR MULTIPLE PATHOGENS ARE THE RULE RATHER THAN THE EXCEPTION. ATYPICAL IMMUNE RESPONSES IN THE CONTEXT OF COINFECTION MAY NOT ONLY AFFECT THE OUTCOME OF DISEASE BUT ALSO DIAGNOSIS AND THERAPY. OUR AIM IS TO UNDERSTAND THE IMPACT OF CONCURRENT PARASITIC INFECTIONS SUCH AS MALARIA ON TUBERCULOSIS. TO DO SO, WE COMBINE THE MOUSE MODEL OF AEROGENIC TUBERCULOSIS WITH DIFFERENT INFECTION MODELS FOR MALARIA IN ORDER TO INVESTIGATE THE MUTUAL INFLUENCE OF BOTH DISEASES IN THE COINFECTED HOST. MOST IMPORTANT FINDINGS Tuberculosis (Tb) and malaria are the most prevalent bacterial and parasitic infections in humans, respectively and continue to be major causes of morbidity and mortality in impoverished regions in the tropics. Not only are both conditions important threats to public health on their own, but in combination may promote severe disease and mortality more than either disease alone. Malaria is known to suppress the immune system and to impair immune responses to unrelated pathogens and vaccines. Accordingly, malaria is associated with secondary infections and malaria prevention leads to a decrease in death numbers that exceeds malaria mortality reduction. A recent study carried out in Guinea-Bissau indicates that malaria Figure 1: A) Immunological events upon Mtb-PbNK65 coinfection (working model). B) BMMΦ produce high amounts of IL-10 and support Mtb survival in the presence of pRBCs prevention can reduce mortality in severely ill Tb patients. Ro- tions and to antagonize pro-inflammatory responses essential dent models of malaria-Tb coinfection revealed impaired con- for protective immunity to Mtb. In vitro we found increased trol of mycobacterial infections in the presence of parasites. IL-10 production and impaired IFN-γ mediated growth control In our experimental models, we use different natural rodent of BCG by macrophages concurrently exposed to parasitized malaria parasites which recapitulate the variety in clinical ma- red blood cells (pRBCs) in contrast to those exposed to BCG nifestations of human malaria to answer the question if and alone (Fig. 1 B). how both severe and self-limiting malaria infections interfere Self-resolving malaria and Tb with the control of Mtb in the coinfected host. In order to study how a mild malaria infection interferes with Severe malaria and Tb Tb we use the non-lethal strain of P. yoelii (Py) which allows us We recently showed that coinfection with Plasmodium berghei to follow the consequences of coinfection until the very late NK65 (PbNK65) rendered mice more susceptible to Mtb infec- chronic phase of Mtb infection. While the course of Py infec- tion as chronic Tb was exacerbated. PbNK65 coinfection was tion is unchanged during coinfection, Mtb numbers increase. associated with aggravated lung inflammation, hypercytokine- Interestingly, the numbers of CD11c+ cells are elevated in the mia, dysregulated T cell responses, and increased mycobacte- presence of Py in lung and spleen. Isolated CD11c+ cells from rial replication. Highly elevated cytokine levels in tissues and Py infected spleens exposed to Mtb ex vivo internalize more sera together with severely increased and uncontrolled rec- bacteria and promote their growth better compared to CD11c+ ruitment of leukocytes to the lungs of coinfected mice indicate cells from naïve mice. The underlying mechanisms and the in an overly aggressive pro-inflammatory immune response and vivo relevance of this observation will be further investigated. impaired immune regulation (Fig. 1 A). Despite high levels of Concurrent Mtb infection interferes with live attenuated ma- IFN-γ and TNF-α, coinfected mice were unable to control Mtb laria vaccine efficacy replication presumably due to high levels of IL-10 which is Coinfection with Mtb and PbNK65 exacerbates chronic Tb but known to deactivate macrophage antimicrobial effector func- it renders mice more resistant to malaria. Reduced suscepti- Seite 41 / PRA Infections MAZEN ATIA • JANNIKE BLANK • LARS EGGERS • SIMON GOTTWALT • NADINE HARMEL bility is reflected by decreased parasitemia and ameliorated impaired immune control of infection. Together, these studies liver pathology compared to mice infected with PbNK65 alo- indicate that Mtb coinfection causes modulation of the im- ne. Moreover, we found that the initial occurrence of parasi- munological environment and impedes plasmodial liver-stage tes in the blood (prepatency) following PbNK65 sporozoite mediated protection. Given the high prevalence of Tb in ma- transmission is delayed in animals pre-infected with Mtb, in- laria endemic areas, this would have major implications on dicating impaired liver-stage development when Mtb is con- whole-sporozoite vaccination approaches. current. While this can be of advantage for disease outcome and severity, it might have opposite effects on the efficacy of SELECTED PUBLICATIONS whole-sporozoite vaccination approaches. Immunization with Schneider BE, Behrends J, Hagens K, Harmel N., Shayman JA attenuated sporozoites which are arrested during liver-stage and Schaible UE. (2014) Lysosomal phospholipase A2: A novel development confers sterile protection against subsequent player in host immunity to Mycobacterium tuberculosis. Eur. J. Plasmodium challenge in mouse and man. It has been propo- Immunol., 44: 2394–2404. doi: 10.1002/eji.201344383 sed that the persistence of a memory T cell response depends at least in part on the persistence of an antigen-depot in the Mueller AK, Behrends J, Blank J, Schaible UE and Schneider liver. Our data suggest that the presence of Mtb interferes BE. (2014) An Experimental Model to Study Tuberculosis-Ma- with parasite liver load and with vaccine efficacy. In fact, we laria Coinfection upon Natural Transmission of Mycobacteri- found that mice are less well protected against wild type (WT) um tuberculosis and Plasmodium berghei. J. Vis. Exp. (84), sporozoite challenge after experimental immunization with e50829, doi:10.3791/50829. genetically attenuated parasites (GAPs; Δuis3) when Mtb is concurrent. We immunized mice according to a prime-2-boost Mueller AK, Behrends J, Hagens K, Mahlo J, Schaible UE and regime which confers 100 % protection against subsequent Schneider BE. (2012) Natural Transmission of Plasmodium Plasmodium WT challenge (Fig. 2). However, immunity was berghei Exacerbates Chronic Tuberculosis in an Experimental hampered in mice inoculated with Mtb two days after the last Co-infection Model. PloS ONE 7(10): e48110. immunization, indicated by one mouse that lost protection and developed blood stage parasitemia (Fig. 2, Table). This INTERNAL AND EXTERNAL COLLABORATIONS finding was confirmed in a second experiment and further Internal collaborations: corroborated by a significant increase in parasite liver load in Ulrich Schaible, Division of Cellular Microbiology a group of mice sacrificed 42 h after WT challenge (Fig. 2, Jochen Behrends, Core Facility Fluorescence Cytometry Liver load). The GAP vaccine elicits immunity against parasite Gabriele Schramm, Experimental Pneumology liver stages, thus an increased liver load corresponds to an External collaborations: Ann-Kristin Müller, University Hospital Heidelberg Thomas Jacobs, Bernhard Nocht Institute for Tropical Medicine, Hamburg GRANT SUPPORT Leibniz Graduate School Model Systems for Infectious Diseases: “Malaria – Tuberculosis Coinfection: Dissecting the immunological interactions during concurrent Plasmodium and Figure 2: Protection against WT Plasmodium sporozoites after GAP immunization is impaired upon exposure to Mtb. Liver load 42 h after WT challenge is significantly increased when mice are exposed to Mtb after vaccination and protection from blood-stage infection is partially abrogated. Mycobacterium infection” IMMUNOBIOPHYSICS • PD DR. ANDRA SCHROMM LUNG SURFACTANT MACROPHAGE ACTIVATION AND REGULATION MEMBRANE BIOPHYSICS IMMUNE MODULATORS MISSION THE DIVISION OF IMMUNOBIOPHYISCS IS FOCUSED ON THE INITIATION AND REGULATION OF THE INNATE IMMUNE RESPONSE OF MACROPHAGES TO VIRULENCE FACTORS DERIVED FROM BACTERIAL MEMBRANES. IN AN INTERDISCIPLINARY APPROACH, WE AIM AT UNDERSTANDING PROCESSES INVOLVED IN THE IMMUNE DEFENSE AGAINST INFECTIOUS DISEASES ON THE MOLECULAR LEVEL. OUR RESEARCH IS BASED ON A TOOLBOX OF STATE OF THE ART METHODS INCLUDING RECONSTITUTION SYSTEMS FOR LIPID MEMBRANES, BIOPHYSICAL TECHNIQUES TO INVESTIGATE MEMBRANE PROPERTIES, CONFOCAL MICROSCOPY, HIGH SENSITIVITY SPECTROSCOPY, AND MODERN IMMUNOLOGICAL AND CELL BIOLOGICAL TECHNIQUES. MOST IMPORTANT FINDINGS Legionella pneumophila derived outer membrane vesicles engage a unique mechanism to deliver pathogen factors to host cells The formation and release of outer membrane vesicles (OMVs) is a phenomenon observed in many bacteria, including Legionella pneumophila, the causative agent of Legionaires disease. During infection this human pathogen primarily invades alveolar macrophages. The shedding of OMV by L. pneumophila is observed in intro and in vivo. In a collaboration with the Division of Clinical and Experimental Pathology, it could be demonstrated that OMV cause severe histological Figure 1: Entry of L. pneumophila OMV in macrophages. Left panel, damage in human lung tissue explants. To characterize and binding (black line) and uptake (blue line) of OMV and E. coli understand the mode of action of these pathogen derived bioparticles. Right panel, OMV in human macrophages visualized by membrane vesicles, we performed biophysical analysis of confocal microscopy (scalebar 25 µm). the membrane properties of OMVs. Using the highly brilliant phila OMVs on human macrophages. Our data support OMV X-Ray source at the German Electron Synchrotron DESY we membrane fusion as a new mechanism for the remote delive- could elucidate the membrane architecture of L. pneumo- ry of virulence factors to host cells. phila OMVs. Biophysical investigations into the interaction of OMVs with model membranes demonstrated the incorporati- Role of surfactant lipids in LPS-induced pulmonary inflammation on of OMV membrane material into liposomes composed of Neonatal acute respiratory distress syndrome (nARDS) is a eukaryotic lipids, revealing an endogenous property of OMVs severe inflammatory syndrome in newborns with an activated to fuse with eukaryotic membranes. Furthermore, we inves- immune response of alveolar macrophages, plasma infiltrati- tigated the interaction of OMVs with human macrophages. on, leukocyte extravasation as well as ongoing degradation of Co-incubation experiments showed dose- and time-dependent lung surfactant leading to defective gas exchange and irrever- binding of fluorophore-labeled OMVs to macrophages. Trypan- sible fibrosis of lung tissue. In preterm-born babies requiring blue quenching experiments disclosed a rapid temperature- mechanical ventilation, nARDS caused by bacterial infection independent internalization of OMVs into macrophages at is a frequent complication leading to irreversible lung injury 37 °C and 4 °C, whereas the uptake of E.coli bioparticles by and a high risk of mortality. Type II alveolar cells secrete sur- phagocytosis was restricted to physiologic temperature (Fig. factant as a lipoprotein complex that is essential for surface 1, left panel). OMV could be visualized by confocal microscopy tension as well as lung immunity. Secreted phospholipase A2 at the cytoplasmic membrane and in the cytoplasm of human (sPLA2) is upregulated in alveolar macrophages during nARDS macrophages (Fig. 1, right panel). Purified OMV induced TNFα and catalyzes the hydrolysis of surfactant glycerophospholi- production in macrophages. In summary, we demonstrated pids. In a collaborative project with the Pediatric Clinic, UKSH binding, internalization and biological activity of f L. pneumo- Kiel, the Institute of Immunology, University of Kiel, and the Seite 43 / PRA Infections IRINA VON CUBE • DR. FRANZISKA KOPP • SARAH KUPSCH • MAJED MANSOURI • YVONNE SIEBKEN • MARIESSA STADEMANN • PHILIP TRAUSCHIES Division of Experimental Pneumology, we are investigating the Institute of Physics, University of Lübeck. Analysis of the possibilities to i) modulate the inflammatory immune respon- mechanisms underlying HDP-based regulation of macrophage biology is essential to a targeted development of this class of immune response modifiers. SELECTED PUBLICATIONS Pupo E, Lindner B, Brade H, Schromm AB. Intact rough- and smooth-form lipopolysaccharides from Escherichia coli separated by preparative gel electrophoresis exhibit differential biological activity in human macrophages. FEBS JOURNAL 2013 280(4):1095-1111. Keese SP, Brandenburg K, Roessle M, Schromm AB. Pulmonary surfactant protein A induced changes in the molecular conformation of bacterial deep-rough lipopolysaccharide lead to reduced activity on human macrophages. INNATE IMMUNITY 2014 20(8):787-98. Figure 2: Attenuation of the immune response of human macropha- Jäger J, Keese S, Roessle M, Steinert M, Schromm AB. Fusion ges to LPS by pulmonary surfactant lipids (SLM, reconstituted of Legionella pneumophila outer membrane vesicles with eu- surfactant lipid mixture). karyotic membrane systems is a mechanism to deliver patho- se of the lung (Fig. 2) and ii) to reduce surfactant degradation gen factors to host cell membranes. Cell Microbiol. 2014 Nov by the application of anionic phospholipid species in clinical 3. doi: 10.1111/cmi.12392. [Epub ahead of print] surfactant therapy. Together with the Division of Bioanalytical Chemistry, we are establishing protocols for the identification INTERNAL AND EXTERNAL COLLABORATIONS of lipid species in BAL and surfactant from clinical samples. Internal collaborations: Heinz Fehrenbach, Division of Expe- The analysis of clinical samples is accompanied by in vitro in- rimental Allergology; Torsten Goldmann, Division of Clinical vestigations. These studies are supposed to give insights into and Experimental Pathology; Thomas Gutsmann, Division of the mechanisms of pulmonary immune modulation and may Biophysics; Christian Herzmann, Clinical Trial Center; Uwe Ma- provide a rationale for therapeutic strategies towards nARDS. mat, Division of Structural Biochemistry; Dominik Schwudke, Division of Bioanalytical Chemistry Reprogramming of Macrophage Biology by Host defense External collaborations: Christian Hübner, Lübeck; Martin Krau- Peptides (HDP) se, Clinic of Pediatrics, Kiel; Manfred Roessle, HASYLAB c/o Antimicrobial peptides, also referred to as host-defense pep- DESY, EMBL, Hamburg; Stefan Schütze, Institute of Immuno- tides are effector molecules of the innate immune system logy, Kiel; Michael Steinert, Technical University, Braunschweig instrumental in the immune response to bacterial infection. This class of defense molecules is produced in all epithelial GRANT SUPPORT surfaces and in the lung. Besides their potent antimicrobial DFG/EXC 306 O TP4 Exzellenzcluster „Inflammation at Interfaces “ activity HDP also have the potential to control immune cell Project “Surfactant therapy in neonatal acute respiratory dis- functions, a property of importance in the regulation of over tress syndrome” shooting immune responses such as sepsis or the resolution of chronic inflammatory diseases such as COPD. Basis of this DFG/EXC 306 RA3 Exzellenzcluster „Inflammation at Inter- activity is among others a direct interaction of HDPs with host faces “ cells (macrophages). The effects of HDPs on host cells are in- Project “Anti-inflammatory regulation of immune cells by vestigated in collaboration with the Division of Biophysics and membrane active host defense peptides” INFECTION IMMUNOLOGY • DR. CHRISTOPH HÖLSCHER TUBERCULOSIS MICE INTERLEUKIN-13 GRANULOMA NECROSIS INTERLEUKIN-17 MULTIFUNCTIONAL T CELLS MISSION INFECTION WITH MANY INTRACELLULAR PATHOGENS SUCH AS MYCOBACTERIUM TUBERCULOSIS (MTB) RESULTS IN PERSISTENCE OF THE PATHOGEN AND CHRONIC DISEASE. THE IMMUNE SYSTEM OF THE HUMAN HOST IS CAPABLE OF MOUNTING AN ADEQUATE CELL-MEDIATED INFLAMMATORY IMMUNE RESPONSE AGAINST MTB. HOWEVER, DURING PERSISTING CHRONIC INFECTIOUS DISEASES CELL-MEDIATED INFLAMMATORY REACTIONS MAY ALSO BE DETRIMENTAL TO THE HOST LEADING TO REACTIVATION AND PATHOLOGY. THEREFORE, THE INFECTION IMMUNOLOGY RESEARCH GROUP WANTS TO UNDERSTAND THE CYTOKINE-MEDIATED REGULATION OF (I) PROTECTION AND (II) PATHOLOGY IN ORDER TO DISSECT PROTECTIVE AND PATHOLOGY-PROMOTING MECHANISMS ASSOCIATED WITH THE INFLAMMATORY RESPONSE DURING TUBERCULOSIS (TB). MOST IMPORTANT FINDINGS exact mechanisms how multifunctional T cells are induced by (i) Interferon-gamma (IFN-γ) and interleukin (IL)-17A are two pro- IL-17A and which mechanism expressed by these cells in fact totype T helper (Th) 1 and Th17 cytokines, respectively that play promote protection (Figure 1). a critical role in protection against infections with intracellular pathogens. As we have previously shown, mice lacking IL-23 or (ii) Until today, factors influencing the course of Mtb infection IL-17A production are highly susceptible to infection identifying are only incompletely defined. One important reason is the fact an IL-23-dependent Th17 immune reaction as an important ef- that genetically or immunologically tractable mouse models fector arm of protective immune responses. On the other hand, do not exist that displays the characteristic features of human Th17 cells also play an essential role in the development of chro- granulomas in TB patients: centrally necrotizing lesions, a strict nic inflammatory diseases, especially those mediated by T effec- stratification of a fibrous capsule that separates the necrotizing tor cells. This proverbial function of IL-17A as a “double-edged granuloma from the adjoining tissue, foamy macrophages found sword” is highlighted by our studies of TB in IL-27 receptor-alpha adjacent to the fibrous capsule within the necrotic lesion, and (Rα)-deficient mice. In these mice, an enhanced inflammatory most importantly hypoxia. Because necrotizing granulomas and immune response to Mtb infection results in a better control the associated hypoxia might be a key factor in the pathogenesis of mycobacterial growth but also lead to immunopathology of post-primary TB and eventually the distribution and success and premature death. We could now show that these opposed of Mtb, the Infection Immunology research group developed a effects are exclusively mediated by an elevated expression of mouse model in which Mtb infection results in granuloma ne- IL-17A that leads to an increased frequency of IFN-γ-TNF-IL-2- crosis strongly resembling the pathology of human TB. In these expressing multifunctional T cells. Currently, we examine the IL-13-overexpressing (tg) mice, arginase-1-expressing alternatively activated macrophages, which have previously shown by us to promote susceptibility to intracellular pathogens, drive the typical pathology with centrally necrotizing granulomas with a fibrous rim and foamy macrophages (Figure 2). A key attribute of lesions in post-primary TB that we have also found is hypoxia, which may have a great impact on gene expression and the metabolic activity of Mtb strains within lesions of infected individuals. In granulomas from TB patients and Mtb-infected IL-13tg mice, a zone of lipid-rich, acid-fast bacilli-containing foamy macrophages are found adjacent to the fibrous capsule within the necrotic lesion. In these foamy macrophages, the lipid metabolism of the host and of Mtb are believed to play a central role in persistence and Figure 1: Model of the IL-17A-mediated recruitment of multifunctional T cells. granuloma necrosis. Together, the pathology observed in Mtbinfected IL-13tg mice display many features of human post-prima- Seite 45 / PRA Infections MAHIN ABAD DAR • MARTINA ACKERMANN • DR. JOCHEN BEHRENDS • JULIA BÖHME • DR. HANNA ERDMANN • ALEXANDRA HÖLSCHER • MIRIAM KRUSCH • KRISTINA RITTER • GABRIELE RÖVER • ANNA STUBBE • KERSTIN TRAXEL • FILIPA VARELA • JOHANNA VOLZ • DR. KERSTIN WALTER • MELANIE WANNICK ry TB. This mouse model is therefore an ideal tool to study the INTERNAL AND EXTERNAL COLLABORATIONS progression of tuberculosis and to determine factors important Ehlers S, Research Center Borstel; Schaible U, Division of for the clinical outcome. It can help to define host determinants Cellular Microbiology, Research Center Borstel; Niemann S, involved in creating a niche for the replication and survival of the Molecular Mycobacteriology, Research Center Borstel; Peter- bacteria in the host under physiologcial stress to eventually iden- sen F, Biochemical Immunology, Research Center Borstel; tify factors that are involved in tissue pathology during post pri- Breloer M, Lotter H, Jacobs T, Bernhard-Nocht-Institute for Tropical Medicine, Department of Immunology, Hamburg, Germany; Berod L, Sparwasser T, Institute of Infection Immunology, TWINCORE, Centre for Experimental and Clinical Infection Research, a Joint Venture between the Medical School Hanover and the Helmholtz Centre for Infection Figure 2: Features of human post-primary TB in Mtb-infected IL-13tg mice. Research Braunschweig, Germamary TB and trigger granuloma formation and necrosis which do ny; Lang R, Molecular Medicine, University Hospital Erlangen, not only contribute to the pathology of the disease but are also Germany; Köhl J, Institute for Systemic Inflammtion Research, involved in transmission. Moreover, this model will serve as a tool University of Lübeck; Brombacher F, International Centre for to identify mycobacterial factors such as glycolipids that promote Genetic Engineering and Biotechnology, University of Cape granuloma necrosis as an exit strategy Town, South Africa. SELECTED PUBLICATIONS GRANT SUPPORT Behrends J, Renauld JC, Ehlers S, Hölscher C. IL-22 is mainly BMBF; KMU-innovativ-5:Verbundprojekt: „Struktur-basiertes produced by IFNγ-secreting cells but is dispensable for host Desing trypanozider Wirkstoffe, Teilprojekt B“ protection against Mycobacterium tuberculosis infection. BMBF; Nation-wide Collaborative Grant: „Pulmonary Tuber- PLOS ONE 2013; 8: e57379. culosis – host and pathogen determinants of resistance and disease progression.“ Workpackage E (Animal models). Heitmann L, Schoenen H, Ehlers S, Lang R, Hölscher C. Minc- BMBF; DZIF „Thematic Translational Unit – Tuberculosis“: TTU le is not essential for controlling Mycobacterium tuberculosis 02.702 „MycoMouse“ and TTU 02.802 „Preclinical Test Station“ infection. IMMUNOBIOL 2013; 218: 506-516. DFG; IRTG1911: “Immunoregulation of Inflammation in Allergy and Infection”: TPB6 „Regulatory T cell modulated vaccina- Heitmann L, Abad Dar M, Schreiber T, Erdmann H, Behrends tion against tuberculosis“ J, Mckenzie ANJ, Brombacher F, Ehlers S, Hölscher C. The IL- DFG; GRK 1727: „Modulation von Autoimmunität“ : Projekt A4 13/IL-4R-alpha axis is involved in tuberculosis-associated pa- “Modulation of experimental autoimmune encephalitis by the thology. J PATHOL 2014; 234: 338 regulatory T cell-derived cytokine interleukin-35” Universität zu Lübeck; FSP „Modulation von Infektion und All- Berod L, Stüve P, Varela F, Behrends J, Swallow M, Kruse F, ergie“: Projekt B2 „Die Rolle von Komplement bei der Infekti- Krull F, Ghorbani F, Mayer CT, Hölscher C*, Sparwasser T*. on mit Mycobacterium tuberculosis“ Rapid rebound of the Treg compartment in DEREG mice limits Universität zu Lübeck; FSP „Modulation von Infektion und the impact of Treg depletion on mycobacterial burden, but Allergie“: Projekt B3 „Untersuchung der optimalen Zytokin- prevents the development of autoimmunity. PLOS ONE 2014; vermittelten T und B Zell-Antworten nach der Impfung gegen 9:e102804 (*equal contribution). Mycobacterium tuberculosis“ MICROBIAL INTERFACE BIOLOGY • PD DR. NORBERT REILING MYCOBACTERIUM TUBERCULOSIS MACROPHAGE PATHOGEN VARIABILITY PHAGOSOME ISOLATION WNT SIGNALING LIPID METABOLISM IN VITRO DRUG TESTING MISSION THE DIVISION OF MICROBIAL INTERFACE BIOLOGY FOCUSES ON THE DETAILED MOLECULAR CHARACTERIZATION OF THE INTERACTION BETWEEN PATHOGENIC MYCOBACTERIA AND THEIR TARGET CELLS, THE MACROPHAGES. THIS INCLUDES DETAILED STUDIES ON (I) PATHOGEN VARIABILITY TO UNDERSTAND THE MACROPHAGE RESPONSE TO MYCOBACTERIUM (M.) TUBERCULOSIS CLINICAL ISOLATES, (II) THE STRUCTURAL AND FUNCTIONAL CHARACTERISATION OF PATHOGEN-CONTAINING PHAGOSOMES, (III) THE FUNCTIONAL ROLE OF WNT6 IN M. TUBERCULOSIS INFECTION AND (IV) THE DEVELOPMENT AND USE OF NOVEL IN VITRO SCREENING SYSTEMS TO IDENTIFY ANTI-MYCOBACTERIAL COMPOUNDS MOST IMPORTANT FINDINGS Lipid-based, immunomagnetic isolaton and characterization Clade-specific virulence patterns of clinical isolates of the of M. tuberculosis phagosomes Mycobacterium tuberculosis Complex To identify structural differences between MTBC phagoso- Clinical strains of the Mycobacterium tuberculosis complex mes we established a novel lipid-based, immunomagnetic (MTBC) are genetically more diverse than previously anticipated. method to isolate and functionally characterize M. tubercu- We have identified Clade-specific virulence patterns of clinical losis–containing phagosomes from primary host cells. Elec- isolates of the MTBC in human primary macrophages and ae- tron microscopic and biochemical analyses of the magnetic rogenically infected mice. M. tuberculosis lineages, also termed phagosome-containing fractions provided evidence of an en- clade I, comprising “modern” lineages, such as Beijing and Euro- hanced presence of bacterial antigens and a differential distri- American Haarlem strains, showed a significantly enhanced ca- bution of proteins involved in the endocytic pathway over time pability to grow in human macrophages (Fig. 1) compared to clade as well as cytokine-dependent changes in the phagosomal II strains, which include “ancient” lineages, such as, e.g., East Afri- protein composition. Due to its relative speed and versatility, can Indian or M. africanum strains. Our data reveal three different the magnetic isolation procedure facilitates the comparative pathogenic profiles: ((i) Beijing strains are characterized by low biochemical and mass spectrometric analysis of M. tubercu- uptake, low cytokine induction, and a high replicative potential, losis-containing phagosomes (Clade I vs Clade II). This should (ii) Haarlem strains by high uptake, high cytokine induction, and promote the identification of essential cellular factors and mechanisms, which are needed to successfully eradicate M. tuberculosis by its host cell. Wnt6, a novel macrophage-derived immunoregulatory player in M. tuberculosis infection We have recently identified a regulatory role of Wnt proteins in inflammatory and infectious disease settings including tuberculosis. We now show that Wnt6 is expressed in granulomatous lesions of M. tuberculosis infected mice and is involved in macrophage differentiation and proliferation. We identified foamy macrophage-like cells as the primary source of Wnt6 in the infected lung and uncovered a TLR-MyD88-NF-kappaB dependent mode of induction in macrophages. Functional Figure 1: Differential growth of MTBC strains in human macropha- studies in M. tuberculosis-infected macrophages uncovered ges. Fold growth (day7 vs day1) of MTBC clade I and clade II strains a Wnt6-dependent induction of Arginase-1 and the downregu- in human macrophages (Reiling et al. mBio 2013). lation of TNF-alpha, which points to an unexpected novel role high growth rates, and (iii) EAI strains by low uptake, low cytokine for Wnt6 in macrophage differentiation shifting polarization induction, and a low replicative potential. Future studies addres- towards an M2 phenotype. These findings and our observati- sing the underlying mechanisms and clinical implications need to on that the majority of Wnt6 expressing cells (Fig. 2) contain take into account the diversity of both the pathogen and the host. lipid vesicles, suggest that M. tuberculosis induces Wnt6 to Seite 47 / PRA Infections JULIUS BRANDENBURG • SVENJA GOLDENBAUM • VICTORIA HORNS • KATHARINA KOLBE • LISA NIWINSKI • KATRIN SEEGER • DR. CHRISTINE STEINHÄUSER promote the formation of foamy macrophages as a cellular S, Schneider-Brachert W, Schütze S, Reiling N. Lipid-labe- habitat to persist and replicate within the host. ling facilitates a novel magnetic isolation procedure to characterize pathogen-containing phagosomes. Traffic. 2013 Mar;14(3):321-36. Steinhäuser C, Dallenga T, Tchikov V, Schaible UE, Schütze S, Reiling N. Immunomagnetic isolation of pathogen-containing phagosomes and apoptotic blebs from primary phagocytes. Curr Protoc Immunol. 2014 Apr 2;105:14.36.1-14.36.26. Schaale K, Brandenburg J, Kispert A, Leitges M, Ehlers S, Reiling N. Wnt6 is expressed in granulomatous lesions of M. tuberculosis-infected mice and is involved in macrophage differentiation and proliferation. J Immunol. 2013 Nov 15;191(10):5182-95. Figure 2: Expression of Wnt6 in the lung of M. tuberculosis–infected mice. Formalin-fixed lung sections of M. tuberculosis H37Rv–infected mice [1000 CFU; day 42 p.i.] (Schaale et al. J. Immunol 2013). Michelucci A, Cordes T, Ghelfi J, Pailot A, Reiling N, Goldmann O, Binz T, Wegner A, Tallam A, Rausell A, Buttini M, FastScreen: Screening Systems of the Identification of novel Linster CL, Medina E, Balling R, Hiller K. Immune-responsive anti-TB compounds gene 1 protein links metabolism to immunity by catalyzing ita- Our macrophage expertise has prompted us to screen the conic acid production. Proc Natl Acad Sci U S A. 2013 May drug efficacy of novel anti-TB compounds in M. tuberculo- 7;110(19):7820-5. sis-infected primary macrophages. Compounds of interest are identified in a newly developed 96 well based medium INTERNAL AND EXTERNAL COLLABORATIONS throughput system using GFP-expressing M. tuberculosis, •K Brandenburg, N Gisch, T Goldmann, G Graßl, T Gutsmann, which allows the screening of small to medium compound lib- C Herzmann, C Hölscher, C Lange, B Lindner, S Niemann, E raries for anti-TB activity. In parallel putative cytotoxic effects Richter, U Schaible, D Schwudke on primary macrophages are measured online in the same •D Adam, T Lindhorst, S Schütze, S Sebens, A Tholey, Univer- format. All three systems have been successfully used and sity of Kiel; J Rupp, University of Lübeck; K-H Wiesmüller, are embedded within the thematic translational transfer unit EMC microcollections, Tübingen; E Krause, FMP, Berlin; G Tuberculosis (TTU-Tb) within the “Deutsches Zentrum for In- van Zandbergen, Paul Ehrlich Institute, Langen; T Pukrop, fektionsforschung (DZIF)”. University of Göttingen; I Rosenkrands, Statens Serum Institut, Copenhagen (DK); M Sweet, IMB, Brisbane (Aus), P SELECTED PUBLICATIONS Walther, University of Ulm; M. Lerm, Linköping University (S); Reiling N, Homolka S, Walter K, Brandenburg J, Niwinski L, K Hiller, Luxembourg Centre for Systems Biomedicine (L) Ernst M, Herzmann C, Lange C, Diel R, Ehlers S, Niemann S. Clade-specific virulence patterns of M. tuberculosis complex GRANT SUPPORT strains in human primary macrophages and aerogenically in- •DFG Priority Program 1580 “Intracellular Compartments as fected mice. MBio. 2013 Jul 30;4(4). pii: e00250-13. Places of Pathogen-Host-Interaction”(Re1228/5-1 and Re 1228/5-2) Steinhäuser C, Heigl U, Tchikov V, Schwarz J, Gutsmann T, Seeger K, Brandenburg J, Fritsch J, Schroeder J, Wiesmüller KH, Rosenkrands I, Walther P, Pott J, Krause E, Ehlers •DFG Cluster of Excellence 306 ”Inflammation at Interfaces”, Project “Lipid Disorders” •BMBF Grant 01KI0784,” TB or not TB”, WP D. MODELS OF INFLAMMATION • PROF. DR. GUNTRAM GRAßL SALMONELLA FIBROSIS INFLAMMATION GLYCOSYLATION INFECTION IBD MISSION OUR GASTROINTESTINAL TRACT HAS A HUGE SURFACE AREA WHICH IS EXPOSED TO A MYRIAD OF MICROBES AND MICROBIAL ANTIGENS. WHILE IT HAS TO TOLERATE HARMLESS COMMENSAL BACTERIA, IT NEEDS TO MOUNT AN APPROPRIATE IMMUNE REACTION TO PATHOGENIC BACTERIA. AN ABERRANT IMMUNE RESPONSE CAN LEAD TO CHRONIC INFLAMMATION AND DEVELOPMENT OF INFLAMMATORY BOWEL DISEASE (IBD). USING IN VIVO AND IN VITRO INFECTION MODELS, WE TRY TO UNDERSTAND: I. HOW ENTERIC PATHOGENS – ESPECIALLY SALMONELLA ENTERICA SEROVARS TYPHIMURIUM AND INFANTIS – INTERACT WITH THE HOST EPITHELIUM AND FIBROBLASTS TO INITIATE INTESTINAL INFLAMMATION AND FIBROSIS II. HOW THE IMMUNE SYSTEM RESPONDS TO ACUTE AND CHRONIC INFECTION WITH BACTERIA MOST IMPORTANT FINDINGS gut and increased histopathological changes for the emergent Emergence of Salmonella Infantis pESI containing strains. Therefore, pESI plays a key role in the Diseases caused by Salmonella enterica serovars are a major successful spread of S. Infantis in a short time of only 2-3 ye- health burden world wide. To date, there are more than 2500 ars (see Figure 1; Aviv et al., Environ Microbiol, 2014). serovars known. Most of them cause mild to severe gastroen- Role of NOD2 in Salmonella-triggered inflammation: teritis (e.g. S. Typhimurium) or typhoid fever (e.g. S. Typhi). Until S. Typhimurium ΔmsbB contains a modified lipid A, contai- recently Salmonella serovars Enteritidis and Typhimurium ac- ning penta- and hexa-acylated lipidA, which is a poor agonist counted for the majority of gatrointestinal infection. However, for TLR4. The ΔmsbB mutant has reduced virulence in mice. in several countries including Israel and Hungary, serovar In- NOD-like receptors (NLRs) are pattern recognition receptors fantis has become one of the most commonly isolated Salmo- for bacterial ligands. The NLR NOD2 is a cytoplasmic receptor nella serovars in patients with severe gastrointestinal symp- for muramyl-dipeptide (a part of the bacterial cell wall). Loss- toms. By comparative analyses between pre-emergent and the of-function mutations NOD2 are risk factors for developing clonal emergent S. Infantis populations we identified a mosaic Crohn’s disease which is characterised by chronic intestinal megaplasmid, designated pESI, with an approximate size of inflammation. How NOD2 mutations result in un-controlled 280 kb. Strains containing the pESI have increased resistance inflammation in the gut is not well understood. We showed to heavy metals and are also multi-drug resistant to antibiotics that infection of NOD2-deficient and NOD2+/+ control mice such as tetracycline, sulfamethoxazole and trimethoprim. Fur- with wildtype Salmonella Typhimurium leads to comparable colonization and intestinal inflammation. In contrast, infection with the attenuated mutant Salmonella Typhimurium ΔmsbB leads to increased inflammation in NOD2-/- mice compared to controls. In vitro transfection of PRRs into HEK cells showed that S. Typhimurium ΔmsbB triggers decreased proinflammatory cytokine production through TLR4 but strongly increased proinflammatory cytokine production through TLR2. Thus exa- Figure 1: Emergence of Salmonella Infantis: A. Cases of clinical cerbated inflammation in NOD2-deficient mice infected with Salmonella isolates is Israel from 1996 to 2012. B. Pathology score S. Typhimurium ΔmsbB is likely due to altered NOD2-TLR cros- of streptomycin-pretreated C57BL/6 mice that were orally infected stalk (Claes et al., PLOS One, 2014). with S. Infantis and the virulence plasmid containing strain S. Infantis Salmonella and intestinal glycosyltransferases pESI. Note the increased inflammtion triggered by the pESI containing strain (Aviv et al., Environ Microbiol, 2014). Glycans on mucosal surfaces play an important role in host-microbe interactions. Human blood groups are among the oldest thermore pESI containing strains have enhanced adhesion known genetic polymorphisms, although their existence and and invasion into mammalian and avian cells and in vivo ex- possible adaptive value remain largely enigmatic. It is hypo- periments demonstrated enhanced colonization of the mouse thesized that blood groups exist as a byproduct of pathogen- Seite 49 / PRA Infections JANIN BRAUN • ANNE-KATHRIN CLAES • ANTJE CORNELIUS • LEONIE DREWS • DOROTHEE SCHULTZ • JANICE SEIDEL • DR. NATALIE STECK • STEPHANIE STEIN • ABDULHADI SUWANDI • BASTIAN REBIEN driven selection, particularly in the gastrointestinal tract where Stress’. 2014. Springer, ISBN 978-3-642-30017-2 blood group-related genes are also highly expressed, although only limited evidence for this hypothesis exists. We tested the Mojibian M, Lam AW, Fujita Y, Asadi A, Grassl GA, Dickie P, hypothesis that differences in susceptibility to enteric patho- Tan R, Cheung AT, Kieffer TJ. Insulin-Producing Intestinal K gens contribute to the maintenance of expression variation at Cells Protect NOD Mice from Autoimmune Diabetes. Gastroen- the blood group-related gene B4galnt2 in house mice. In sup- terology. 2014 Mar 21. doi: 10.1053/j.gastro. 2014.03.020 port of this hypothesis we find that the removal of B4galnt2 glycans from the intestinal mucosa results in significantly Aviv G, Tsyba K, Steck N, Salmon-Divon M, Cornelius A, Ra- decreased pathology upon experimental infection with Sal- hav G, Grassl GA, Gal-Mor O. A unique megaplasmid contri- monella Typhimurium. This is supported by detailed histopa- butes to stress tolerance and pathogenicity of an emergent thological analysis of intestinal tissue and the measurement Salmonella enterica serovar Infantis strain. Environ Microbiol. of pro-inflammatory markers post-infection. Further, extensive 2014;16(4):977-94 metagenomic analysis of fecal bacterial communities before and after infection which were done in collaboration with the INTERNAL AND EXTERNAL COLLABORATIONS group of John Baines (CAU Kiel, MPI Plön), demonstrate a role INTERNAL: Norbert Reiling: Wnt signalling in gut and lung in- of B4galnt2 genotype-dependent changes in the intestinal mi- flammation, Ulrich Zähringer, Holger Heine: TLR4 and NOD2 crobiota in susceptibility to S. Typhimurium (see Figure 2). crosstalk, Helmut Haas, Gabi Schramm: Schistosoma-Salmonella co-infections. Ulrich Schaible: Role of LPLA2 in Salmonella infections. Ulrich Schaible, Norbert Reiling, Dieter Adam: Lysosomal disorders and bacteria-induced inflammation EXTERNAL: John Baines, MPI Plön & CAU Kiel: Gut microbiota in fibrosis; Philip Rosenstiel, Simone Lipinski, CAU Kiel: Inflammation in NLR deficient mice; Jürgen Brinckmann, University Figure 2: Glycosylation of Mucin-2 in the mouse intestine. Co-staining of Lübeck: Extracellular matrix composition in fibrosis; Ohad against with DBA (Dolichus Biflorus Aggluttinin) lectin staining and Gal-Mor: Molecular and cellular characterization of Salmonel- antibodies against MUC2 reveals MUC2 as a potential target for la enterica serovar Infantis pathogenicity; Dan Littman, Gret- B4galnt2 glycosylation. Green: DBA-lectin (N-Glycans), red: MUC2, chen Diehl, Skirball Institute, New York, USA and Brett Finlay, blue: DAPI (nuclei). UBC Vancouver, Canada: Th17 response in gut fibrosis SELECTED PUBLICATIONS GRANT SUPPORT Claes AK, Steck N, Schultz D, Zähringer U, Lipinski S, Rosen- DFG Priority Program SPP1656: ‘The role of blood group-rela- stiel P, Geddes K, Philpott DJ, Heine H, Grassl GA. Salmonella ted glycosyltransferases in shaping diversity of the intestinal ΔmsbB triggers exacerbated inflammation in Nod2 deficient microbiota and susceptibility to inflammation’” mice. PLoS One. 2014 Nov 25;9(11):e113645 DFG: Excellence Cluster 306 “Inflammation at Interfaces”: ‘Lysosomal disorders and bacteria-induced inflammation’ Gal-Mor O, Boyle EC, Grassl GA. Same species, different di- The German Israeli Foundation for Scientific Research and De- seases: how and why typhoidal and non-typhoidal Salmonella velopment (G.I.F): “Molecular and cellular characterization of enterica serovars differ. Front Microbiol. 2014. 4;5:391 Salmonella enterica serovar Infantis pathogenicity” Steck N, Grassl GA. Free radicals and pathogens – role for reactive intermediates in innate immunity. Book chapter in ‘Systems Biology of Oxidative Stress Systems Biology of Oxidative MOLECULAR MYCOBACTERIOLOGY • PROF. DR. STEFAN NIEMANN MYCOBACTERIUM TUBERCULOSIS COMPLEX MOLECULAR EPIDEMIOLOGY POPULATION STRUCTURE PATHOBIOLOGY EVOLUTION VIRULENCE MISSION OUR RESEARCH IS FOCUSED ON A BETTER UNDERSTANDING OF THE EPIDEMIOLOGY OF TUBERCULOSIS (TB) IN LOW AND HIGH INCIDENCE SETTINGS, ON THE ANALYSIS OF THE GLOBAL POPULATION STRUCTURE, GENOMIC DIVERSITY, PATHOBIOLOGY AND VIRULENCE OF MYCOBACTERIUM TUBERCULOSIS COMPLEX (MTBC) STRAINS, AND ON THE INVESTIGATION OF MOLECULAR DETERMINANTS AND MICROEVOLUTION OF RESISTANT STRAINS. MOST IMPORTANT FINDINGS were analyzed tor performance tests and determination of pa- Epidemiology, population structure and microevolution of resis- rameters for genome based molecular epidemiology/phylogeny. tant strains. High resolution typing of bacteria and easy identifi- Whole genome sequencing (WGS) subdivided the outbreak into cation of emerging clones exhibiting increased resistance and/or several genome clusters and unique SNP profiles (Fig. 1). A ma- transmissions rates is essential for epidemiological surveillance ximum of three SNPs was identified in eight confirmed human- and disease control. For MTBC isolates, typing based on MIRU- to-human transmission chains. We estimated the MTBC genome VNTR (mycobacterial interspersed repetitive units variable num- evolutionary rate at 0.4 mutations per genome per year. As a ber tandem repeat) analysis represents the current standard. standardized, portable, and expandable approach for genome However, with the development of “next generation sequencing based MTBC surveillance, we developed and evaluated core technologies“ (NGS), the analysis of near-complete genomes of genome multi locus sequence typing (cgMLST). The cgMLST ap- clinical isolates for e.g. resistance detection or tracing outbreaks proach (3,041 genes) discriminated 26 outbreak strains with a has become possible. resolution comparable to that of classical SNP-based WGS typing For efficient analysis of clinical isolates, we established a flexib- and is currently evaluated with larger strain sets. WGS sequenci- le pipeline enabling NGS analysis from low DNA amounts (one ng was also applied to discern the evolution of multidrug resistant ng) for smaller strain sets e.g. for diagnostic questions (Illumina (MDR) strains under long-term therapy in the patient, as well as MiSeq BenchTop Sequencer) or larger strain collections e.g. for the global population structure and evolutionary history of the population based studies (HiSeq 2500/NextSeq 500). 86 strains MTBC. Here, we used global strains collections as well as archeo- with identical classical genotyping patterns from a longitudinal logical samples from AD 1028 to AD 1280 to characterize the outbreak detected in the city of Hamburg from 1997 to 2010 global diversity and to reconstruct the evolutionary history of the Figure 1: Minimum pathogen. Coalescent analyses based on 279 genomes indicate spanning tree allowing that the MTBC emerged about 70,000 years ago, accompanied hypothetical nodes of the migrations of anatomically modern humans out of Africa, and ex- M.tuberculosis outbreak in Hamburg and Schleswig- panded as a consequence of increases in human population den- Holstein. The year of sity during the Neolithic period. This long co-evolutionary history isolation is coded by color. is consistent with MTBC lineages displaying characteristics indi- HH, Hanseatic City of cative of adaptation to both low and high host densities. Based Hamburg; SH, Schleswig- on comparative analysis of three 1,000-year-old mycobacterial Holstein. Asterisk indicates the root of the tree determined through compa- genomes from Peruvian human skeletons, we demonstrated that a member of the MTBC caused human disease in the Americans rison with an outgroup before contact with the New World. Using two independent dating (H37Rv) (Roetzer et al. approaches, the data obtained suggest a significantly different PLoS Med 2013 evolutionary scenario with the most recent common ancestor of doi:10.1371/journal. the MTBC existing less than 6,000 years ago, which supports a pmed.1001387.g001). Holocene dispersal of the disease. Resistance mechanisms. Resistance in MTBC strains is based on the emergence of variants in the genomes (mainly single nucleotide polymorphisms, SNPs) representing ideal targets for ef- Seite 51 / PRA Infections PATRICK BECKERT • DOREEN BEYER • DR. ANNA ENGSTROEM • DR. SUSANNE HOMOLKA • DR. SILKE FEUERRIEGEL • DR. CHRISTIANE GERLACH • LEILA JELJELI • DR. THOMAS KOHL • ANJA LÜDEMANN • GLENNAH KERUBI • SILVIA MAASS • DR. SVEN MALM • DR. MATTHIAS MERKER • VANESSA MOOR • ECATERINA NOROC • DR. JUDITH PETERSEN • TANJA STRUWE-SONNENSCHEIN • DR. BARBARA TIZZANO • TANJA UBBEN • JULIA ZALLET ficient detection of resistances in clinical isolates. Applying WGS, SELECTED PUBLICATIONS the near-complete detection of resistance mechanisms in a given Bos KI, Harkins KM, Herbig A, Coscolla M, Weber N, Comas isolate has become reality, possibly guiding individualized therapy I, et al. Pre-Columbian mycobacterial genomes reveal seals of TB patients in the near future. However, this requires a tho- as a source of New World human tuberculosis. Nature 2014 rough understanding of the functional impact of genomic variants 23;514(7523):494-7. in approx. 35 target regions described to be involved in resistance development. Accordingly, we continued our work on resistance Comas I, Coscolla M, Luo T, Borrell S, Holt KE, et al. Out-of- mechanisms and performed a large-scale multicenter study on Africa migration and Neolithic coexpansion of Mycobacte- pyrazinamide resistance determinants interrogating 1,950 clini- rium tuberculosis with modern humans. Nat Genet 2013 cal isolates. Overall, 280 genetic variants were identified, which 45(10):1176-82. could be divided into four classes: (i) very high confidence resistance mutations (85 %), (ii) high-confidence resistance (3 %), Miotto P, Cabibbe AM, Feuerriegel S, et al. Mycobacterium (iii) mutations with an unclear role (2 %), and (iv) mutations not tuberculosis pyrazinamide resistance determinants: a multi- associated with phenotypic resistance (10 %) (Fig. 2). We further center study. MBio 2014 21;5(5):e01819-14. verified the impact of natural genome variation in potential resisReiling N, Homolka S, Walter K, et al. Clade-specific virulence patterns of Mycobacterium tuberculosis complex strains in human primary macrophages and aerogenically infected mice. MBio 2013 30;4(4). pii: e00250-13. Roetzer A, Diel R, Kohl TA, et al. Whole genome sequencing versus traditional genotyping for investigation of a Mycobacterium tuberculosis outbreak: a longitudinal molecular epidemiological study. PLoS Med 2013 10(2):e1001387. INTERNAL AND EXTERNAL COLLABORATIONS Internal collaborations: Bioanalytical Chemistry, Infection Immunology, Microbial Interface Biology, Cellular Microbiology, Figure 2: Distribution of genetic variants across the four categories identified: (i) very high confidence resistance mutations, (ii) high-confidence resistance mutations, (iii) mutations with an unclear role, and (iv) mutations not involved in phenotypic resistance. The number of isolates Fluorescence Cytometry, Diagnostic Mycobacteriology, Clinical Infectious Diseases, Bioinformatics, IT External collaborations: Harmsen D, Münster; Kalinowski J, belonging to each category is also reported. The inner ring shows the Bielefeld; Nübel N, Braunschweig; Gagneux S, Basel; Rhode percentages of mutations affecting the structure of the enzyme for each K, Florida; Supply P, Lille; Wirth T, Paris. category of genetic variants. PZA-R, PZA resistance. *, including Networks: German Center for Infection Research, TBPANNET wild-type isolates for the pncA gene (Miotto et al. Mbio 2014). Consortium, PathoNgenTrace Consortium, Global Microbial tance genes by analyzing sequence diversity in 20 resistance- Identifier Consortium. associated genes in a collection of 71 strains encompassing the major MTBC lineages. Overall, 58 SNPs were found, further con- GRANT SUPPORT firming that it is of outstanding importance to distinguish phyloge- Our work is supported by several grants of the German Re- netically informative neutral polymorphisms from true resistance- search Foundation, the Federal Ministry of Education and Sci- conferring mutations for accurate molecular based diagnostics. ence, the EU-FP7. Currently, larger strain collections are being analyzed by WGS with the final aim to develop a validated resistance SNP encyclopedia. BIOCHEMICAL IMMUNOLOGY • PROF. DR. FRANK PETERSEN MAST CELL-NEUTROPHIL INTERACTION ASTHMA AUTOIMMUNITY PROTEASES TISSUE DAMAGE MISSION THE WORK OF THE DIVISION IS FOCUSED ON THE ANALYSIS OF PATHOPHYSIOLOGICAL PROCESSES IN THE EFFECTOR PHASE OF CHRONIC INFLAMMATORY PROCESSES IN THE LUNG AND SKIN. WE ARE INVESTIGATING THE REGULATORY PRINCIPLES AND MECHANISMS UNDERLYING THE INTERACTION OF MAST CELLS AND NEUTROPHILS IN THE PATHOGENESIS OF ASTHMA AND AUTOANTIBODY-MEDIATED INFLAMMATION. MOST IMPORTANT FINDINGS blistering diseases. Until now, protease activities are determined Asthma manifests as a chronic inflammation of the airways in supernatants or tissue extracts by measuring the cleavage of which is characterized by reversible airway constriction, in- colorimetric or fluorescing substrates. In a cooperation initiated filtration of inflammatory cell in to the lung, increased mucus and funded by the German Center for Lung Research (DZL) with production and airway remodeling. Two major processes of di- Carsten Schultz from the EMBL in Heidelberg we established sease transition, from ‚healthy to diseased‘ and from ‚controlled a new technique using cell bound protease sensors. Protease asthma to aggravated disease‘, have been recently identified as activity can be monitored on single cell level by a change in the major fields of research in the Priority Areas Asthma & Allergy. change of the fluorescence resonance energy transfer (FRET) Within this context, our division focused on the analysis of the in- between two fluorophores in the substrate. Due to the complexi- terplay between mast cells and neutrophils with the epithelium ty of the models in experimental in asthma and COPD, we deci- and T cells in the effector phase of allergic asthma. We postula- ded to using the more simple models of autoantibody-mediated te a mast cell – neutrophil axis which represents a fundamental skin blistering disease to establish this methodology. In this pathophysiological element essentially involved in the execution study we could demonstrate a relationship between neutrophil and regulation of the asthma pathology. adhesion and site-specific protease activity. In contrast to acti- Increased numbers of neutrophils in the lung are characteristic vated non-adherent cells, adherent neutrophils displayed high for asthma especially during late-phase reactions and in cases protease activity in the presence of an excess of macromole- of severe and steroid-resistant forms of the disease. However, cular protease inhibitors (Fig. 1). The tight adhesion creates a since in earlier reports it has been shown that in the classical Th2-driven mouse models of asthma the main features of the allergic lung inflammation developed independently from the presence of neutrophils, the pathophysiological role of these cells has long been underestimated. We could show that depletion of neutrophils results in a significant reduction of the airway hyperreactivity and airway inflammation as well as of the goblet cell hyperplasia and mucus production indicating that neutrophils are essential for the pathogenesis of asthma. Unexpectedly, we found high amounts of IL-17 in diseased mice in the absence of neutrophils. IL-17 represents a key player in the orchestration of the neutrophil defense against pathogens. On the other hand, this cytokine plays a critical role autoimmunity, and, as shown most recently, also in Asthma and COPD. Our data provide first evidence that neutrophils are able to inhibit the production of Figure 1: „Hidden“ elastase enzyme activity detected by the FRET probe NEmo-2 on immune complex (IC)-activated neutrophils in presence of the elastase inhibitor alpha1-antitrypsin. Adherent neutrophils display elastase activity at the attachment side which is resistant to protease inhibition.. IL-17 in the inflamed tissue. protected space, a secluded microenvironment between cell Neutrophil proteases such as elastase have been identified as and substratum which excludes proteinase inhibitors and allow relevant mediators of tissue damage in the lung more than 30 a high local concentration of effector molecules (Fig. 2). Brea- years ago and uncontrolled protease activity represents a seve- king the ‚closed space‘ of neutrophils adherent to tissue could re problem in COPD and asthma but also in autoimmune skin facilitate the access of intrinsic as well as therapeutic inhibitors Seite 53 / PRA Asthma and Allergy MARJAN AHMADI • REZA AKBARZADEH • DR. NESTOR GONZALEZ ROLDAN • CHRISTINE ENGELLENNER • HANNO EWERS • CINDY HASS • DIANA HEINRICH • GABRIELE HUSS • DR. BRIGITTE KASPER • JUNIE TCHUDJIN MAGATSIN • DR. SANDRA MINGE • CAROLA SCHNEIDER • DR. XINHUA YU SELECTED PUBLICATIONS Zheng J, Petersen F & Yu X. 2014, ‚The role of PTPN22 in autoimmunity:Learning from mice.‘ AUTOIMMUNITY REVIEWS, Bd 13, Nr. 3, S. 266-271., 10.1016/j.autrev.2013.10.011. Bermbach S, Weinhold K, Roeder T, Petersen F, Kugler C, Goldmann T, Rupp J & König P. 2014, ‚Mechanisms of CiliaDriven Transport in the Airways in the Absence of Mucus‘ AMERICAN JOURNAL OF RESPIRATORY CELL AND MOLECU Figure 2: The protected space hypothesis of neutrophil-mediated LAR BIOLOGY. In press tissue damage. Activated neutrophils release ROS and elastase, which will be immediately deactivated by inhibitors and scavengers in Yu X, Zheng J, Collin M, Schmidt E, Zillikens D & Petersen F. non-adherent cells. Adhesion prevents the access of inhibitory 2014, ‚EndoS Reduces the Pathogenicity of Anti-mCOL7 IgG molecules to proteinases and ROS enabling uncontrolled activity of through Reduced Binding of Immune Complexes to Neutro- these mediators leading to tissue damage. at the site of tissue damage and, therefore, could open a new phils‘ PLOS ONE, Bd 9, Nr. 2, S. e85317., 10.1371/journal. pone.0085317. therapeutical window for targeting the disease. An acute allergic reaction initiated by the activation of mast cells INTERNAL AND EXTERNAL COLLABORATIONS is followed by the subsequent infiltration of many inflammatory Mast cell-neutrophil interaction in asthma - O. Holst, B. Lind- cells, predominantly neutrophils and eosinophils, into the lung ner, Research Center Borstel; J. Köhl, ISEF, P. König, Institute tissue. Based on this observation, we hypothesize a cross talk of Anatomy, M. Kopp, M. Weckmann, Department of Pediat- between mast cells and neutrophils in allergic lung inflamma- ric Allergy and Pulmonology, University of Lübeck; S. Hogan, tion. Beside the characterization of indirect immune regulatory Cincinnati Children‘s Hospital. S100A8/A9 proteins in asth- processes mediated by proteases, we presently identify lung ma and autoimmunity – T. Vogl, University of Münster. FRET mast cell-derived mediators directly involved in the recruitment protease reporters – T. Scholzen, Research Center Borstel; and activation of neutrophils and eosinophils. Comparable to C. Schultz, EMBL Heidelberg. Autoimmunity - D. Zillikens, R. the immediate and delayed reaction in the allergic response Ludwig, Institute of Dermatology, University of Lübeck; G. Rie- we could show that primary human lung mast cells activated mekasten, Charité Berlin. by FcεRI-crosslinking release chemotactic mediators in two phases. A first peak is observed after 30 min of activation and neu- GRANT SUPPORT trophil and eosinophil chemotaxis we determined the lipid medi- DFG SFB/TR 22 “Allergische Immunantworten der Lunge“, ator LTB4 a the as the only relevant chemotaxin here. After 24h Teilprojekt A11. of stimulation, a second activity peak becomes visible. Impor- DFG Cluster of Excellence „Inflammation at Interfaces”; tantly, chemotactic activity is associated with the presence of a Research Area H: IRN Autoimmunity to Type VII Collagen. 30-50 kDa candidate protein, which does not belong to a group DFG GRK 1727 „Modulation von Autoimmunität“ Teilprojekt 12 of previously described mast cell-derived neutrophil/eosinophil DFG IRTG 1911 „Immunoregulation of Inflammation in Allergy activators. Currently, we purify this protein and we will subject and Infection“ the material to proteome analysis. After identification, this medi- BMBF Deutsches Zentrum für Lungenforschung, ARCN ator will be evaluated in biomaterials (sputum and lavage) from asthma and COPD patients available from the biobank of the DZL. According to the two major research fields of the Priority Area, we hope to define a novel biomarker suitable for asthma phenotyping or for the prediction disease progression CLINICAL AND MOLECULAR ALLERGOLOGY • PROF. DR. UTA JAPPE FROM CLINIC RESEARCH AND BACK TO THE CLINIC TO MISSION •CLARIFICATION OF PATHOMECHANISMS OF ALLERGY AND PUTATIVE SENSITIZATION ROUTES •IDENTIFICATION OF NEW ALLERGY-RELEVANT EPITOPES (FROM NATURAL SOURCES OR BIOLOGICAL DRUGS) •SUBSEQUENT DETECTION OF BIOMARKERS FOR RISK MANAGEMENT, DISEASE AND TREATMENT MONITORING TRANSLATIONAL ASPECTS •ASCERTAINING THE ACQUISITION OF WELL-DEFINED PATIENT SUBGROUPS •ESTABLISHING REGISTERS FOR SOURCES OF ALLERGENS AND NEW ALLERGENS •PROVIDING NEW ALLERGENS FOR COMPONENT-RESOLVED DIAGNOSIS •PERFORMING CONFIRMATORY PROVOCATION TESTS WHICH IS POSSIBLE DUE TO THE CLOSE ASSOCIATION TO TWO ALLERGY OUTPATIENT CLINICS, HEADED BY PROF. DR. JAPPE MOST IMPORTANT FINDINGS Are lipophilic house dust mite allergens associated with asthma? Novel lipophilic peanut allergens House dust mites (HDM) are one of the common causes for Peanut is one of the most hazardous food allergen sources. respiratory allergies. Only three single allergens of Dermato- Since allergy diagnostic testing is based on aqueous extracts, phagoides pteronyssinus (Der p) and two of Dermatophagoi- potential allergenic proteins of lipophilic nature are usually un- des farinae (Der f) are available for routine allergy diagnostics. derrepresented or even absent. Because of the high content Especially with regard to lipophilic allergens a lot of informati- of lipids in peanut, the focus is on the purification of hydropho- on is still lacking which is due to their low concentration in the bic/lipophilic allergens to determine their clinical relevance. respective diagnostic extracts or because they are hidden in a A novel strategy to isolate all known peanut oleosins simulta- complex matrix. However, there is evidence from other allergen neously was developed. sources (i. e. peanut) that lipophilic allergens are associated with severe clinical reactions. Some hydrophobic HDM-allergens have already been identified (e. g. Der p 5, 7, 13, and 14). For studying these allergens in more detail as probable cause for HDM-asthma and for improving diagnostic tests, we started Figure 1: New peanut allergens Further separation of the different oleosins was realized by a single run on a preparative electrophoresis cell (Figure 1). All known peanut oleosins were identified by mass spectrometry and N-terminal sequencing, including the allergens Ara h 10, Ara h 11, the presumed allergen oleosin 3, and additional oleosins variants. These oleosins are immunologically distinct. When used as target proteins in Western blotting experiments with sera of patients with mostly severe peanut allergy, binding of IgE-antibodies documented their allergenicity. Figure 2 Seite 55 / PRA Asthma and Allergy MARISA BÖTTGER • MAREN HOHN • DR. ARNE HOMANN • DR. SKADI KULL • DR. SANDRA MINGE • CAROLIN MURAWSKI • PROF. DR. ARND PETERSEN • DANIEL ROSERO • SIMONE ROSS • ALEXANDRA SCHARF • DR. FRAUKE SCHOCKER • MARION SCHULDT • CHRISTIAN SCHWAGER • DANIELA WARNEKE • AUSZUBILDENDE: JANNA MÜLLER; ANN-CHRISTIN BERGMANN; CAROLIN GOLIN; DOMINIC TÖDT; KHIRA SCHILD • GÄSTE: MILICIA GROZDANOVIC, BELGRAD, SERBIEN (2013 DAAD) JASNA NIKOLIC (DAAD-STIPENDIATIN AUS BELGRAD, SERBIEN, 2014) with the expression of some lipophilic HDM-allergens in E. coli INTERNAL AND EXTERNAL COLLABORATIONS as well as in Pichia pastoris. In parallel, we used well characte- Inhouse: PD Dr. A. Frey: Epitope mapping; microarray development; rized patients’ sera for the assessment of individual sensitiza- Prof. Dr. T. Goldmann: Interactions between allergens and membra- tion patterns on the basis of the recombinant HDM-allergens nes; PD Dr. Lindner & Dr. Schwudke: Characterization of allergens Der p 1, 2, 4, 5, 7, 10, 11, 14, 15, 18, 21, and 23 (some pro- by mass spectrometry; Prof. Dr. Gutsmann: Functional microbio- vided by S. Vrtala, Vienna), applying immunoblot analysis in logical studies on legume defensins; Prof. Dr. Holst; Dr. Duda, Dr. order to identify marker allergens for HDM-asthma (Figure 2). Gonzales: (Glyco-) Lipids in the allergic reaction (VIP-project) IgE to certain epitopes as biomarker for drug allergy risk as- National: Prof. Dr. D. Kabelitz (Institute for Immunology, UK SH, sessment Campus Kiel) and PD Dr. A. Kromminga (IPM Biotech GmbH, Therapeutic monoclonal antibodies show promising effects on Hamburg): IgE-binding epitopes on biologicals; Prof. Dr. M. Eh- widespread, but, up to now, not effectively treatable diseases. lers (ISEF; University of Lübeck) Glycosylation pattern of IgG Reports are increasing in number that biologicals can induce (SPP A5); Prof. Dr. B. Przybilla, (LMU, Munich): Allergens re- IgE-mediated allergic reactions and/or antibody-mediated neu- levant for delayed anaphylaxis; Prof. Dr. Simon & Prof. Dr. R. tralisation of therapeutic effects. Foreign peptide sequences as Treudler (Dept. of Dermatology, University of Leipzig) Molecular well as glycans have been described to be responsible for these treatment of food allergy effects which in general lead to a discontinuation of the treat- International: Prof. Dr. J. Baumert (University of Nebraska, ment and/or the switch to another type of therapeutic antibody. Lincoln, USA): Detection of peanut allergens/traces by ELISA However, the treatment alternative sometimes does not prove to technique; Prof. Dr. M. Gavrovic-Jankulovic (University of Bel- be safe. In our study, sera from those patients were investigated. grade, Serbia): structural investigation‘s on food allergens; The oligopeptide binding characteristics along with structure- Dr. J. Lidholm (Uppsala, Sweden): Peanut and lupine allergy; function analysis point to a possible IgG-mediated neutralisation Dr. A. Mari, Rome, Italy Molecular Allergology; Prof. Dr. T.A.E of the biologicals. Serum analysis yields an individual anti-bio- Platts-Mills (Asthma and Allergic Diseases Center, University logical profile for each patient. Taken together, these analyses of Virginia, USA): cross-reactive carbohydrate determinants shall lead to a knowledge-based allergenicity assessment of (alpha-GAL); allergy to biologicals; Prof. Dr. S. Vrtala (University each type of biological and to a suggestion for a safer clinical the- of Vienna) House dust mite allergy rapy switch (risk management of target treatments) in the future. SELECTED PUBLICATIONS GRANT SUPPORT Special research field (SFB)/Transregio 22: „Allergic immune Kleine-Tebbe J, Jappe U (eds). Molekulare Allergologie: Einfüh- responses of the lung“, Project Z1 (B. Lindner, A. Petersen) rung mit kommentierten Kasuistiken. Wissensakademie Dus- DFG Scho 828/2-1 Untersuchung von Erdnussallergenen in tri Verlag 2014 ISBN 978-3-87185-491-0 der Muttermilch (F. Schocker) DFG JA1007/2-1 und DFG PE 491/10-1 Struktur-und Funkti- Petersen A, Rennert S, Kull S, Becker WM, Notbohm H, Gold- onsuntersuchungen von lipophilen Erdnussallergenen, insbe- mann T, Jappe U. Roasting and lipid binding provide allerge- sondere den Oleosinen (U. Jappe) nic and proteolytic stability to the peanut allergen Ara h 8.Biol Member of the German Center for Lung Research (DZL), Airway Chem. 2014 Feb;395(2):239-50 Research Center North (ARCN), Disease Area: Asthma and Allergy, AA-2.2 Cellular mechanisms (U. Jappe) Becker WM, Jappe U. Peanut allergens. Chem Immunol Allergy Bundesministerium für Wirtschaft und Technologie (BMWi) 2014; 100:256-67. doi: 10.1159/000359916. Epub 2014 May 22. KF2784701AJ0: AiF ZIM Entwicklung von Tests für IgE gegen therapeutische monoklonale Antikörper (Uta Jappe) Kleine-Tebbe J, Jappe U. Molecular allergy diagnostic tests: Bundesministerium für Bildung und Forschung (BMBF) Ver- development and relevance in clinical practice. Allergologie bundprojekt BASALIT (Birch Associated Soy Allergy and Immu- 2013; 36: 327-349 no-Therapy) (U. Jappe) EXPERIMENTAL PNEUMOLOGY • PROF. DR. HEINZ FEHRENBACH ALLERGIC BRONCHIAL ASTHMA AIRWAY REMODELING AIRWAY EPITHELIUM MAST CELLS BASOPHILS NANOTOXICOLOGY DESIGN-BASED STEREOLOGY MISSION TO CONTRIBUTE TO A BETTER UNDERSTANDING OF THE PATHOGENESIS OF INITIATION AND EXACERBATION OF CHRONIC INFLAMMATORY LUNG DISEASES SUCH AS ALLERGIC BRONCHIAL ASTHMA AS WELL AS THE DEVELOPMENT OF NOVEL PREVENTIVE AND THERAPEUTIC APPROACHES. MOST IMPORTANT FINDINGS ross the Disease Areas and DZL sites. By establishment of Air- Allergic bronchial asthma is one of the most common chronic Liquid-Interface cultures of murine and human alveolar as well diseases in the world. According to the WHO-funded Global as of airway epithelial cells (Fig. 1) we will investigate the role Initiative for Asthma Report on the Global Burden of Asthma of the airway epithelium in asthma both in mice and humans. as many as 300 Million people of all ages and all ethnic back- In close cooperation with the junior research group Mouse grounds suffer from asthma. The tremendous impact of asth- Models of Asthma, the novel interleukin IL-37 was identified ma on health care systems is demonstrated by the annual to be able to ablate a Th2 cell directed allergic inflammatory direct medical and indirect economic costs (ie loss of work response and the hallmarks of experimental asthma in mice, days, lost productivity, premature retirement), which accumu- which suggests that IL-37 may be critical for asthma pathoge- late to approximately € 19.5 Billion for Europe. In Germany nesis. Furthermore, the data suggest a mode of action of IL-37 approx. 368.000 work days per year are reclusively lost due that involves IL-18Rα as well as the orphan receptor SIGIRR/ to asthma ranging almost on the same level like diabetes and IL-1R8. Further, Sina Webering in her PhD studies elucidated chronic-ischemic heart diseases. the role of Th17 cells and of the transcription factor RORγt in The airway epithelium is the initial target of well-known risk the pathogenesis of neutrophilic asthma (Fig. 2). factors (e.g. viruses, cigarette smoke, particles) of airway di- Figure 2: Results of the seases and of trigger factors for disease exacerbations. There- FACS analysis revealed a fore, studying the effects of airborne triggers on the airway significant increase in epithelium is a major focus of the group. In turn, the epithe- Th17 cells in lung tissues lial responses to such triggers translate into immunological in a model of neutrophilic responses. In allergic diseases such as allergic asthma, mast cells and basophilic granulocytes are of particular significance asthma (OVA) versus control mice. and therefore, our studies will additionally address the functional relevance of these cell types. Together with the Clinical Helminth-derived molecules have been identified as a new and Experimental Pathology, we have established the DZL pri- therapeutic approach for various immune-mediated diseases. mary cell culture and tissue laboratory which is an important We aim at developing novel preventive strategies against all- infrastructure of the DZL enabling DZL-wide cooperations ac- ergic diseases such as allergic asthma by making use of the anti-inflammatory potential factors isolated from the eggs of the helminth Schistosoma mansoni. Recently, the administration of Schistosoma mansoni soluble egg antigens was demonstrated to reduce the severity of colitis in an adoptive transfer mouse model characterized by an increased Th2 response and a suppressed Th17 response. Furthermore, Kathrin Knuhr could demonstrate in her PhD studies that IL-4 released from basophilic granulocytes that were stimulated with the egg-derived factor IPSE/alpha-1 inhibits the release of pro-inflammatory cytokines Figure 1: H&E stained section of an air-liquid-interface culture of primary epithelial cells isolated from mouse trachea showing the well-differentiated airway epithelium with ciliated, non-ciliated and basal cells that developed in vitro. such as IL-1β and IL-6 from LPS-stimulated human monocytes. With regard to mast cell biology, special emphasis is given to the degranulation of mast cells in health and allergic disease. Seite 57 / PRA Asthma and Allergy JULIANE ARTELT • FRANZISKA BEYERSDORF • MYLÈNE DIVIVIER • KATHRIN KNUHR • FRAUKE KOOPS • CARINA KRÜTZMANN • KRISTINA LANGHANS • SANDRA NYENHUIS • PATRICIA PRILLA • GESINE RODE • DR. GABRIELE SCHRAMM • KERSTIN VIERTMANN • DR. CHRISTINA VOCK • SINA WEBERING • DR. ZANE ORINSKA In this context, the mast cell intrinsic regulatory elements of Bratu VA, Erpenbeck VJ, Fehrenbach A, Rausch T, Rittinghau- tetraspanin proteins localized in the cell membrane and their sen S, Krug N, Hohlfeld JM, Fehrenbach H. Cell counting in interaction with FceRI-complex and c-kit tyrosine kinase are human endobronchial biopsies--disagreement of 2D versus being investigated. For example, miR-155 was recently de- 3D morphometry. PLoS One. 2014 Mar 24;9(3):e92510. monstrated to play a critical role in FcεRI-mediated mast cell responses by modulating components of the PI3Kγ pathway. INTERNAL AND EXTERNAL COLLABORATIONS Further, tissue-specific characteristics and regulatory mecha- Close collaborations within the Research Center Borstel are nisms of mast cells (e.g. receptor expression, IgE loading, pat- established with the Divisions of Mouse models of Asthma, tern of protease expression, and reactivity) are addressed in Invertebrate Models, Innate Immunity, Mucosa Immunology order to better understand why allergic reactions are more or and Diagnostics, Immunobiophysics and the Division of Clini- less restricted to specific organs and compartments and how cal and Experimental Pathology. this restriction emerges during organ development. This newly Several research projects are pursued in cooperation with identified mechanism of miRNA-controlled MC activation may groups at the University of Lübeck: Peter König, Institute of affect the initiation and maintenance of allergic disorders The Anatomy, and Gereon Hüttmann, Institute for Biomedical Op- initial hypothesis behind this project is that organ specific re- tics; at the University of Kiel: Martin Krause, Department of gulatory mechanisms and/or their deficiency are crucial for Pediatrics University Medical Center Schleswig-Holstein; Ste- the establishment of an allergic immune response (Fig. 3). fan Schütze, Institute of Immunology. Figure 3: Fluorescence Close external collaboration has been established with the part- microscopic demonstration ners of the BMBF funded joint project “NanoCOLT”: Henning of mast cells (TexasRed- Bockhorn (Karlsruhe Institute of Technology), Bernd Müller (Phi- Avidin, red) along blood vessels (anti CD31-Alexa lipps-University of Marburg), Tanja Hansen (Fraunhofer Institute 488, green) and within the of Toxicology and Experimental Medicine, Hannover) and Peter extracellular matrix in the König (University of Lübeck) as well as with various collabora- skin of mice. tors at partner-sites of the German Center for Lung Research: Armin Braun (Fraunhofer Institute of Toxicology and Experimental Medicine, Hannover), Matthias Ochs (Institute for Applied and SELECTED PUBLICATIONS Functional Anatomy, Hanover Medical School), Marcus Mall (De- Lunding L, Webering S, Vock C, Schröder A, Raedler D, partment of Translational Pulmonology, Heidelberg). Schaub B, Fehrenbach H, Wegmann M. IL-37 requires IL18Rα and SIGIRR/IL-1R8 to diminish allergic airway inflam- GRANT SUPPORT mation in mice. ALLERGY 2015 Jan 3. doi: 10.1111/all.12566 BMBF: German Center of Lung Research, co-coordinator of [Epub ahead of print]. Disease Area Asthma/Allergy (H.F.); co-cordinator of Platform Imaging (H.F.) Ghosh D, Mueller GA, Schramm G, Edwards LL, Petersen A, BMBF: Consortium “NanoCOLT” (FKZ 03X0093A), deputy co- London RE, Haas H, Gupta Bhattacharya S. Primary identi- ordinator (H.F.) fication, biochemical characterization, and immunologic pro- DFG: Cluster of Excellence “Inflammation at Interfaces” perties of the allergenic pollen cyclophilin cat R 1. JOURNAL (EXC306-POTP3) (H.F.) OF BIOLOGICAL CHEMISTRY 2014 Aug 1;289(31):21374-85. DFG: „Immunomodulation by IPSE/alpha-1, an excretory/ secretory protein from Schistosoma mansoni eggs“ (SCHR Biethahn K, Orinska Z, Vigorito E, Goyeneche-Patino DA, Mirg- 608/4-1) (G.S.) homizadeh F, Föger N, Bulfone-Paus S. miRNA-155 controls DFG: GRK1727 „Modulation of Autoimmunity“, TP11 “IL-15/ mast cell activation by regulating the PI3Kγ pathway and ana- IL-15Rα as targets for modulation of autoimmune reactions phylaxis in a mouse model. ALLERGY 2014 Jun;69(6):752-62. in CNS” (Z.O.) INNATE IMMUNITY • PROF. DR. HOLGER HEINE INNATE IMMUNITY AND ALLERGY DROSOPHILA MISSION THE MISSION OF THE DIVISION OF INNATE IMMUNITY IS THE INVESTIGATION, ANALYSIS AND CHARACTERIZATION OF ACTIVATION MECHANISMS OF THE INNATE IMMUNE SYSTEM THROUGH MICROBES, MICROBIAL STRUCTURES AND ALLERGENS. IN PARTICULAR, WE EXAMINE THE INTERPLAY AND COMMUNICATION OF EPITHELIAL AND DENDRITIC CELLS WITH EACH OTHER AND THE ADAPTIVE IMMUNE SYSTEM AND THE CONSEQUENCES OF THIS ACTIVATION FOR LUNG DISEASES SUCH AS ALLERGY AND ASTHMA. MOST IMPORTANT FINDINGS thanobrevibacter smithii are known to be part of the indigenous Allergy and asthma: human gut microbiota. Although the immunomodulatory effects Innate intranet communications: modulation of dendritic cell of bacterial gut commensals have been studied extensively in responses by epithelial cells the last decade, the impact of methanoarchaea in human’s The interaction of various types of innate and adaptive immu- health and disease was rarely examined. Whereas exposure ne cells during an ongoing allergic response is pivotal for the to M. stadtmanae leads to substantial release of proinflamm- outcome and the resolve of the disease. Dendritic cells (DCs) atory cytokines in moDCs, only weak activation was detected are key regulators of this network, however, signals derived after incubation with M. smithii. Phagocytosis of M. stadtmanae from other participating components such as the epithelium by moDCs was demonstrated by confocal microscopy (Fig.1) as in turn affect their function. Thus, we studied the direct interaction of DCs with epithelial cells as well as the modulation of DC-functions by soluble modulators from activated human primary bronchial epithelial cells (NHBEs). A co-culture model involving Calu-3 cells, cultured under air-liquid interface conditions, and monocyte-derived DCs (moDCs) was used to address allergy-modulatory effects of epithelialand dendritic cell interactions at the allergen-entry site. While allergen-stimulation of Calu-3 cells per se did not result in an altered release of immune-stimulatory cytokines, Der p 2- or Bet v 1-treatment of moDCs induced a diverging secretion of the cytokines. Interestingly, cytokine release was strongly increased when direct epithelium/DC contact was enabled. To investigate the effect of soluble epithelial factors on DCfunction, we used supernatants of NHBEs that were treated with T cell-derived cytokines such as IL-4, IL-22 or IFN-y and analyzed their modulatory activity on moDCs. Despite the highly differential transcriptional fingerprints of such NHBEs, only marginal changes in secreted inflammatory cytokines of moDCs were detected. Analysis of T cell polarizing factors such as Ox40L showed more condition-dependent expression, but further studies are required to evaluate these effects (supported by BMBF, DZL AA). The intestinal archaea Methanosphaera stadtmanae and Methanobrevibacter smithii activate human dendritic cells (together with Prof. Ruth A. Schmitz, CAU Kiel) The methanoarchaea Methanosphaera stadtmanae and Me- Figure 1: Phagocytosis of M. stadtmanae in moDCs is blocked by Cyt D and crucial for immune cell activation. Lysosomes were stained with LysoTracker Red DND-99. Seite 59 / PRA Asthma and Allergy SUHAD AL-BADRI • KATRIN BÖHNSTEDT • INA GORONCY • DR. ANDRÉ JENCKEL • MANDY MAI • DR. KARINA STEIN (MATERNAL LEAVE) • ANNA STÖRMER • DR. KARIN ULICZKA • PROF. ARTUR J. ULMER well as transmission electronic microscopy (TEM) and shown unidentified pathway of FoxO mobilization independent of the to be crucial for cellular activation by using specific inhibitors. conventional FoxO elicitors (Fig. 2). Both strains, albeit to different extents, initiate a maturation program in moDCs as revealed by up-regulation of the cell-surface receptors CD86 and CD197 suggesting additional activation of adaptive immune responses. Furthermore, M. stadtmanae and M. smithii were capable to alter the gene expression of antimicrobial peptides in moDCs to different extents. Taken together, our findings strongly argue that the archaeal gut inhabitants M. stadtmanae and M. smithii are specifically recognized by the human innate immune system. Moreover, both strains are capable of inducing an inflammatory cytokine res- Figure 2: Hypoxia-induced translocation of FoxO, but not the ponse to different extents arguing that they might have diverse NF-κB-related transcription factors dorsal and relish (yellow arrow: immunomodulatory functions that have been underestimated tracheae; red arrow: nuclear translocation). until now (supported by DFG). Impact of hypoxic conditions on the innate immune system – SELECTED PUBLICATIONS from Drosophila melanogaster to chronic inflammatory disea- Bang C, Weidenbach K, Gutsmann T, Heine H*, Schmitz RA* ses of the lung (together with Prof. Roeder, CAU Kiel) (*equal contribution). The Intestinal Archaea Methanosphae- The innate immune system represents an ancient host defense ra stadtmanae and Methanobrevibacter smithii Activate Hu- mechanism that protects against invading pathogens in verte- man Dendritic Cells. Plos One. 2014;9(6):e99411. brates and invertebrates likewise. Important immune effector molecules to fight microbial infections are antimicrobial pepti- Hogendorf WF, Gisch N, Schwudke D, Heine H, Bols M, Peder- des (AMPs). In the fruit fly Drosophila melanogaster, induction of sen CM. Total Synthesis of Five Lipoteichoic acids of Clostridi- AMP gene expression was thought to depend exclusively on NF- um difficile. Chemistry. 2014;20(42):13511-6. κB transcription factors via activation of two highly conserved signaling cascades, the IMD and the Toll pathway. With respect to Harb H, van Tol EA, Heine H, Braaksma M, Gross G, Over- bacterial or fungal infections this concept is undisputed. Howe- kamp K, et al. Neonatal supplementation of processed su- ver, recent studies show that AMP expression in Drosophila can pernatant from Lactobacillus rhamnosus GG improves aller- also be achieved independently of the canonical NF-κB signa- gic airway inflammation in mice later in life. Clin Exp Allergy. ling pathways and in the absence of a pathogen challenge. One 2013;43(3):353-64. of these non-microbial activators of innate immunity is hypoxia. It occurs particularly during chronic inflammatory disorders e.g. INTERNAL AND EXTERNAL COLLABORATIONS of respiratory organs, where it has a direct effect on the barri- Internal: Otto Holst, Heinz Fehrenbach, Christina Wagner, Uta er epithelium. Depending on the duration and severity, hypoxia Jappe, Nicolas Gisch, Thomas Gutsmann can lead to cell injury and death, and consequently organ injury External: Thomas Roeder, CAU Kiel; Ruth Schmitz-Streit, CAU and failure. From our and other groups’ research, we know that Kiel; Carsten Schmidt-Weber, ZAUM & TU Munich; Peter König, hypoxic stress fulfills a crucial function in various inflammatory University Lübeck ; Alla Zamyatina, BOKU Wien reactions of the epithelia in general. Our intention was to shed light on the connecting mechanisms between hypoxia and inna- GRANT SUPPORT te immunity with focus on the respiratory organs of Drosophila BMBF DZL, disease area AA2.2 melanogaster as a model system. We succeeded in identifying BMBF NanoCOLT, TP4 the forkhead box transcription factor FoxO as a novel activator DFG HE 2758/4-2 of a hypoxia-induced AMP response, and detected a hitherto EKFS 2013-A130 INVERTEBRATE MODELS • DR. CHRISTINA WAGNER DROSOPHILA WOUNDING AIRWAY EPITHELIAL CELLS REMODELLING MACROPHAGES NEUROENDOCRINE BIOGENIC AMINE SYSTEM INNATE IMMUNITY MISSION THE OBJECTIVE OF THE JUNIOR RESEARCH GROUP IS TO ANSWER QUESTIONS WHICH PRIMARILY DEAL WITH THE INNATE IMMUNE SYSTEM AND ITS INVOLVEMENT IN THE PATHOLOGY OF CHRONIC AIRWAY DISEASES SUCH AS BRONCHIAL ASTHMA. USING THE MODEL SYSTEM DROSOPHILA MELANOGASTER WE MAINLY FOCUS ON IMMUNE FUNCTIONS OF AIRWAY EPITHELIAL CELLS AND MACROPHAGES, TWO CELL TYPES, WHICH IN HUMANS ARE MAINLY INVOLVED IN THE DEVELOPMENT, PROGRESSION, AND EXACERBATION OF CHRONIC LUNG DISEASES AND ARE THEREFORE IDEAL TARGETS FOR THERAPEUTIC INTERVENTIONS. MOST IMPORTANT FINDINGS of the current level of the corresponding controls where bas- In more detail the junior research group is interested in indivi- ket expression was not affected. Future mechanistic analysis dual genes, signaling pathways, and effector molecules that will address the question whether further signalling pathways mediate immune reactions and remodelling processes in air- in addition to JNK are involved in FoxO activation to promo- way epithelial cells. In this context genes are especially taken te airway remodelling. In addition, there has been evidence into account of which orthologous have been identified in mice and humans. Such a candidate gene for example represents the longevity and anti-apoptotic gene Fox0 (forkhead box class O), which acts as a nuclear transcription factor. In flies, FoxO is activated in airway epithelial cells during strong immune responses presumably to counteract infection-induced cell death promoting signals and to ensure the survival of the infected epithelial cells. In addition, FoxO also promotes airway remodelling during long-lasting immune reactions that are presumably mediated by the Immune deficiency (Imd) pathway. Due to numerous gain-of-function studies as well as knockout and RNAi studies we could demonstrate that FoxO-induced airway Figure 1: Mechanical sterile wounding induces a systemic immune remodelling affects the entire airway epithelium and is exclu- response in the fruit fly that is characterized by a marked sively characterized by epithelial thickening and proliferation upregulation of antimicrobial peptides (AMPs). of airway epithelial cells. Consequently, we further focused on that FoxO is also involved in aspects of asthma pathology. In the molecular mechanisms that regulate FoxO during airway collaboration with the research groups “Experimental Pneu- remodelling. Additional genetic approaches showed that FoxO mology” and “Mouse Models of Asthma” we could show that is activated by ectopic expression of the IMD-specific pattern FoxO is activated in airway epithelial cells with acute asthma, recognition receptors PGRP-LE and -LC, but not necessarily by whereas in the healthy controls FoxO failed to translocate into downstream-signalling events of the canonical IMD pathway the nucleus and remains inactive. Whether FoxO contributes IKK/Relish. This observation raised the question, which addi- to airway remodelling in mice or not, is yet unclear and will be tional cell signaling pathways linked to IMD might be involved investigated in the upcoming years together with the question in FoxO activation. Just recently obtained results confirmed if FoxO activation affects remodelling in other airway cell types that FoxO might be activated by the signalling pathway c-Jun than epithelial cells of mice such for example smooth muscle N-terminal kinases (JNK), a pathway branching of the Imd cells or fibroblasts. pathway at the level of the TGF-β activated kinase (TAK) and Another important focus of our work is the interaction bet- also regulating FoxO during oxidative stress. Thus, airway epi- ween the neuroendocrine, the respiratory tract, and innate im- thelial cells of transgenic flies, in which the expression level mune system in Drosophila. Here, the essential question is, to of the Jun-N-terminal kinase basket is markedly inhibited via what extent classical stress hormones such as biogenic ami- RNAi, showed a reduction in epithelial thickening at least half nes modulate the innate immune response of macrophages Seite 61 / PRA Asthma and Allergy CARINA KRUETZMANN • STEPHANIE PAPENMEIER and airway epithelial cells. Currently, we focus on the biogenic Quite recently, the junior research group also uses the fruit fly to amines dopamine, serotonin, noradrenaline, and adrenaline investigate individual viral components of human adenoviruses – four neurotransmitters that have their homologs in flies, being associated with acute respiratory diseases in humans. mice, and men. However, so far for both flies and humans it is Here, our emphasis is currently on viral components, which are only known that the adrenergic system modulates the innate involved in immediate early steps of adenovirus infection and immune system in response to stress and therefore, favors which are suspected to interact with components of the host’s at least in humans the exacerbation of chronic lung diseases epithelial immune system. Since in-vivo studies using the model such as bronchial asthma. Right now we investigate the im- system mouse are limited due to low permissivity, the fruit fly pact of all four neurotransmitters on the phagocytic activity with its various possibilities of transgenic manipulation repre- and antimicrobial peptide expression of macrophage-like cells sents an ideal model system to bypass natural infection routes and try to understand, (a) whether G-protein coupled receptor and to validate in-vitro generated data reliably in vivo. signalling pathways modulate phagocytosis in general, (b) to which extend and (c) via which biogenic amine receptors and SELECTED PUBLICATIONS their signalling cascades (eg. adenylate cyclase, phospholipa- Faisal M, Hoffmann J, El-Kholy S, Kallsen K, Wagner C, Bruch- se C) these effects are arranged. haus I, Fink C, Roeder T. (2014) Transcriptional regionalizati- Another stress factor, we focus on, is mechanical wounding on of the fruit fly‘s airway epithelium. PLoS One. 14:9 Roeder T, Isermann K, Kallsen K, Uliczka K, Wagner C. (2011) A Drosophila asthma model - what the fly tells us about inflammatory diseases of the lung. Adv Exp Med Biol. 710:37-47 Wagner C, Isermann K, Roeder T. (2009) Infection induces a survival program and local remodelling in the airway epithelium of the fly. FASEB J. 23:2045-54 Figure 2: Mechanically wounded flies regulate the expression of AMPs (eg. diptericin) via the pattern recognition receptor PGRP-LC (right) and the immune-relevant pathway Imd (left). Wagner C, Isermann K, Fehrenbach H, Roeder T. (2008) Molecular architecture of the fruit fly‘s airway epithelial immune system. BMC Genomics. 9: 446 that effects significantly the systemic immune response of the fruit fly. In this context, we are particularly interested in the INTERNAL AND EXTERNAL COLLABORATIONS pattern recognition receptors, signaling pathways, and effector at the Research Center Borstel we cooperate with die Divisi- molecules involved. So far we were able to figure out that the ons of Innate Immunity (PD Dr. Holger Heine), Experimental immune-relevant signaling pathway Imd but not Toll regulates Pneumology (Prof. Dr. Heinz Fehrenbach), Asthma Mouse Mo- antimicrobial peptide (AMP) expression in response to wound- dels (Dr. Michael Wegmann) ing. Interestingly, wounding evoked by a sterile needle only in- External cooperations has been established with Dr. Sabrina duces the expression of antimicrobial peptides (eg. diptericin) Schreiner Institute of Virology, TUM/Helmholtz Center Munich, which are normally directed against gram-negative bacterial in- Germany and Prof. Dr. Ulrich Theopold, Department of Mole- fections. In case of a damage they are mainly expressed by fat cular Biology and Functional Genomics, Stockholm University, body cells (a functional equivalent of mammalian liver cells), Sweden while hemocytes, homologous to macrophages, do not participate in their production. Which danger signals are released du- GRANT SUPPORT ring the wounding process itself and whether they are related German Research Foundation (DFG): WA 2972 2/2 to those synthesized and released in humans is so far unclear. MOUSE MODELS OF ASTHMA • DR. MICHAEL WEGMANN AIRWAY INFLAMMATION ASTHMA PHENOTYPES LUNG FUNCTION ANIMAL MODELS MISSION TO ESTABLISH MOUSE MODELS OF DIFFERENT ASTHMA PHENO-/ENDOTYPES AND TO USE THESE AS A TOOL TO INVESTIGATE THE PROCESSES AND MECHANISMS UNDERLYING THE PROGRESSION AND CHRONIFICATION OF ALLERGIC AIRWAY INFLAMMATION AND, THUS, THE FORMATION OF THE RESPECTIVE DISEASE. MOST IMPORTANT FINDINGS such a role were provided by the CLARA-study found a signifi- The hallmarks of allergic bronchial asthma comprise a vari- cantly lower expression and production of this new cytokine in able degree of broncho-obstruction, mucus hyperproduction, peripheral blood mononuclear cells from asthmatic children. the developement of airway hyperresponsiveness (AHR) and Local administration of IL-37 to mice with experimental aller- various structural changes of the airway wall summarized gic asthma after allergen provocation significantly down-regu- as airway remodeling. This complex phenotype arises from lated allergic airway inflammation, the expression of various a chronic inflammatory response in the airways. Depending TH2-type and proinflammatory cytokines, mucus hyperproduc- on the severity, type and composition of this immune reaction tion, as well as AHR. Since the receptor and/or co-receptor for several characteristics of asthma experience a special diversification ultimately leading to the formation of different asthma endotypes. Thus, unraveling the mechanisms regulating chronic airway inflammation in asthma is crucial to understand the pathogenesis of asthma. The DZL-funded Junior Research Group “Asthma Mouse Mo- Figure 1: Acute asthma exacerbation is associated with increased dels” establishes mouse models reflecting different endoty- mucus production. PAS-staining of airway cross sections of animals pes of asthma in order to investigate different levels regulating with experimental asthma (OVA), animals with acute asthma airway inflammation. Typically such an inflammation results exacerbation (OVA + pIC) and healthy controls (PBS) to identify tissue destruction, which subsequently triggers the produc- mucus-producing goblet cells. tion of mediators down-regulating the inflammatory reaction IL-37 remains still unknown further investigation will focus on on the hand and inducing repair-mechanisms on the other. elucidating the mode of action of this cytokine in-vivo. We found that patients suffering from allergic bronchial asth- While α-MSH and IL-37 emerged as examples for new regula- ma display significantly enhanced levels of the α-melanocyte tors of inflammation in the pathogenesis of allergic bronchial stimulating hormone (α-MSH), a peptide hormone originally asthma, exogenous triggers such as high allergen load or re- attributed to direct the tanning reaction of the skin. The same spiratory infection have the ability to overwhelm such regu- is true for mice with experimental allergic asthma. Interestin- latory mechanisms and thus lead to acute or chronic aggra- gly, neutralization of α-MSH results in augmented infiltration vation of the disease. Respiratory viral infections are by far of eosinophils into the airway and subsequently increased the most important factor leading to acute exacerbation of mucus production and AHR. Vice versa local application of allergic bronchial asthma, which is associated with dramatic α-MSH markedly reduced airway eosinophilia and improved worsening of asthma hallmarks and the requirement of acute hallmarks of experimental asthma indicating that this peptide medication and/or health care measures. All respiratory vi- hormone exerts anti-inflammatory effects in allergic bronchial ruses that have been associated with such an event (eg rhi- asthma. Such beneficial effects could not be achieved in ani- no viruses, respiratory syncytial virus, influenza viruses, etc.) mals lacking the melanocortin-receptor 5 (MC5-R) expressed use single-stranded (ss) RNA to encode their genome, which on eosinophils and airway epithelial cells suggesting that in turn displays double-stranded (ds) RNA-motifs in its secon- α-MSH requires this receptor to deploy its activity. dary structure or as an intermediate during viral replication. In collaboration with the CPC-M, the DZL-partner side in Mu- Since ds-RNA is recognized intra-cellularly by toll-like receptor nich, IL-37 has been identified to act as a cytokine regulating 3 (TLR-3), we used local activation of TLR-3 by polyIC (pIC) to allergic airway inflammation in asthma. First hints towards model acute exacerbation of experimental allergic asthma in Seite 63 / PRA Asthma and Allergy DR. LARS LUNDING • ALEXANDRA SCHRÖDER • LINDA LANG • STEFFI HAHN mice, which is characterized by dramatically increased mucus is exclusively expressed in TH17 cells and is critical for the hyperproduction and AHR as well as by an inflammatory res- production of IL-17A predisposing it to an promising target for ponse with increased numbers of infiltration eosinophils and therapeutic intervention. Mice that are not able to functionally neutrophils. Using interleukin 17A (IL-17A) deficient mice we express RORγt consequently do not display active TH17 cells could demonstrate that TLR-3-triggered exacerbation of expe- and impaired production of IL-17A. In contrast to wildtype (WT) rimental allergic asthma strictly depends on the availability of mice such mice are nearly refractory towards the induction of IL-17A, which is mainly produced by TH17 cells and NK cells experimental neutrophil asthma underlining the importance in this setting. of these cells and their characteristic transcription factor in this setting. Using small-interference (si) RNA we could demonstrate that downregulation of RORγt in in-vitro generated TH17 cells results in dramatically reduced secretion of IL-17A but also of other proinflammatory cytokines such as IL1β, IL-6 and tumor necrosis factor (TNF) α. Future experiments will focus on an in-vivo strategy to neutralize Th17 cell activities by targeting RORγt in a mouse model of experimental neutrophil asthma. Figure 2: Animals with acute asthma exacerbation display increased SELECTED PUBLICATIONS IL-17 production by TH17 cells and NK cells. FACS analysis of IL-17 Jordakieva G, Wallmann J, Schmutz R, Lemell P, Wegmann producing leukocyte subpopulations of animals with experimental M, Nittke T, Mittlböck M, Fehrenbach H, Godnic-Cvar J, Ziegl- asthma (OVA), animals with acute asthma exacerbation (OVA + pIC) mayer R, Jensen-Jarolim E. Peripheral erythrocytes decrease and healthy controls (PBS). TH17 cells were identified by positive upon specific respiratory challenge with grass pollen allergen staining for CD4 and IL-17, IL-17 producing NK cells by positive staining for NK1.1 and IL-17 and the absence of CD1d-αGalCer. in sensitized mice and in human subjects. PLoS One. 2014 Jan 22;9(1):e86701. Recurrent viral infections and, thus, recurrent acute exacerbations represent the major risk factor for the development INTERNAL AND EXTERNAL COLLABORATIONS of a severe asthma. Typically patients suffering from this At the Research Center Borstel we cooperate with the Divi- asthma endotype display serious impairment of lung func- sions of Biochemical Immunology, Cellular Microbiology, Ex- tion and are highly refractory towards corticoid (CS) therapy, perimental Pneumology, Invertebrate Models and the Fluore- which frequently necessitates intensive medical care, high scence Cytometry Core Facility. External cooperations have dose systemic medication, and mechanical ventilation. Alike been established with the DZL partner sites at the Depart- during episodes of acute exacerbation airway inflammation ment of Pediatric Pneumology & Allergology, University Me- of such patients typically reveals large numbers of infiltrating dical Center Schleswig-Holstein, Campus Centrum Lübeck, neutrophils, which could be the reason for the CS insensitivity. the Department of Pulmonary & Allergy, University Children’s How these neutrophils are initially recruited to the airways of Hospital Munich, LMU Munich, and the Department Transla- these patients remains enigmatic, however, since only in se- tional Pulmonology, University of Heidelberg as well as with vere asthmatics activated TH17 cells have been isolated from the Department of Dermatology, Hospital of the University of inflamed airway tissues, these cells have been hypothesized Münster. to play a major role in the pathogenesis of the disease. We have established a mouse model that mimics key features of GRANT SUPPORT this asthma endotype such as prominent airway neutrophilia, BMBF DZL AA 2.1 infiltration of TH17 cells and AHR to proof this hypothesis. The DFG EXC306-STI transcription factor retinoic-acid orphan receptor γt (RORγt) MUCOSAL IMMUNOLOGY AND DIAGNOSTICS • PD DR. ANDREAS FREY MOLECULAR IMAGING MUCOSAL BARRIERS HIGH THROUGHPUT PEPTIDE ASSAYS MOLECULAR EVOLUTION POINT-OF-CARE DIAGNOSTICS MISSION THE DIVISION OF MUCOSAL IMMUNOLOGY AND DIAGNOSTICS FOCUSSES ITS RESEARCH ON MOLECULAR AND CELLULAR STRUCTURES WHICH MAY SERVE AS MARKERS FOR THE CAUSE, INITIATION, COURSE AND SEVERITY OF MUCOSAASSOCIATED DISEASES, ESPECIALLY IN THE FIELDS OF ASTHMA/COPD AND ALLERGIC DISORDERS. OUR GOAL IS TO DISCOVER SUCH MARKERS, TO CHARACTERIZE AND TO SURVEY THEM FOR DIAGNOSTIC, PROGNOSTIC AND THERAPEUTIC PURPOSES. MOST IMPORTANT FINDINGS cus characteristics in order to diagnose and monitor these The activities of the Division of Mucosal Immunology and Dia- pathologic conditions. A diligent examination of all currently gnostics can be divided into two key sections: available antibodies directed against the major airway mucins The “Diagnostics Section” concentrates on the investigation MUC5AC revealed that none of them is able to detect this mu- of novel diagnostic tools for the analysis of pathological condi- cin in patient samples in a reliable way with little or no inter-pa- tions at mucosal surfaces. One specific topic in this field is the tient variability. Our next task is now the generation of antibo- creation of contrast agents for in-vivo molecular imaging and dies which can capture the major respiratory mucins MUC5AC probes for in-vitro point of care diagnostics. and MUC5B in a patient-independent manner in order to inves- Contrast agents can considerably improve the performance of tigate disease-related alterations of mucin composition of the certain imaging modalities, such as magnetic resonance ima- respiratory tract. ging (MRI) or endoscopy. If a highly specific contrast agent that labels definite target structures or molecules is applied, highlighting of a disease marker down to the (sub)cellular level may be possible, thereby greatly enhancing the diagnostic power of those visualization technologies. In general, a specific contrast agent combines a reporter unit, e.g. a fluorophore or an MRI label, with a specific probe binding unit - the „targeting ligand“ - that can guide the probe to the target of interest. Our research goal is to provide suitable targeting ligands for specific diagnostic purposes on the basis of peptidic molecules. In order to identify peptides that can act as targeting ligands in a given Figure 1: Improvement of the cell penetration efficiency of peptides by molecular evolution. In only 10 rounds of the evolutionary process, the capability of a peptidic carrier to transport fluorescent setting we devised a special strategy for the directed „evolution“ material into life cells was dramatically increased. A: Fluorophore of peptidic molecules. The methodology combines in silico evo- transport into HeLa cells mediated by initial “lead” peptide; B: lution with in vitro testing to quickly obtain promising candidates Fluorophore transport mediated by optimized peptide. Bar, 50 µm. with desired properties. This way a more than 100fold improve- The second, “Immunology” section explores the humoral and ment in target recognition can be achieved. We have successful- structural armament of mucosal barriers and aberrations ly used this approach to enhance the cell penetrating properties thereof which may be involved in pathologic developments. To of peptidic membrane ferries and now have optimized carrier do so we have established several model systems with which systems at hand which can be utilized to transport selected con- we can study the different aspects of relevant interactions on trast agents into distinct cellular compartments. the molecular, cellular and tissue-level. Point-of-care diagnostics for COPD and asthma based on the Elevated proteolytic activity is common for many inflammatory analysis of airway mucins are conceivable since alterations conditions due to an imbalance between proteases and pro- in amount, composition and glycosylation of the respiratory tease-inhibitors. In such environments protease-susceptible mucins appear to reflect different stages and conditions of drugs and diagnostics may be broken down even before they inflammatory lung disorders. We want to develop probes for exert the desired effect. In order to circumvent this obstacle a rapid and diagnostically conclusive analysis of airway mu- toughening of peptides against a proteolytic environment is Seite 65 / PRA Asthma and Allergy DR. BARBARA FREY • DR. MAIK HENKEL • ÖZGE KÖK • DR. THORSTEN KRAUSE • DR. KATRIN RAMAKER • DR. NIELS RÖCKENDORF • JÜRGEN SARAU • DR. HEIKE SINNECKER • ALHEIDIS VON QUAST • GERALDINE WIESE • IMKE WYSOKINSKI 10/2013 necessary. We studied the stabilizing effect of non-canonical Sinnecker H, Ramaker K, Frey A. Coating with luminal gut- amino acids on the proteolytic resistance of peptides towards constitutents alters adherence of nanoparticles to intestinal two prominent inflammation-associated proteases, neutrophil epithelial cells. Beilstein J Nanotechnol 2014, 5:2308-2315. elastase and protease 3, and were able to identify a set of modifications which considerably inreased the stability of model INTERNAL AND EXTERNAL COLLABORATIONS peptides to both enzymes. These findings may pave the way to Inhouse: new peptidic and peptoid agents for the treatment and diagno- Divisions of: sis of chronic inflammatory lung disorders. Barrier Integrity Airborne particles may trigger impairments of the mucosal Biophysics barrier and defense machinery, not only in the airways from Cellular Microbiology which a substantial fraction of inhaled ultrafine matter is Clinical and Experimental Pathology exhaled again but also in the gut where deposited airborne Clinical and Molecular Allergology matter ends up after cilial clearance. Using native gastrointestinal fluids, tissue explants and cultured epithelial lining cells External: we analyzed the interaction of nanoparticles with mucosal DFG-funded research cluster PARENTRY: University of Lübeck; tissues. We observed that mucosal secretions coat synthetic University of Hamburg; University Duisburg-Essen; MPI for Po- nanoparticles by forming a biomolecule corona on the particle lymer Research, Mainz surfaces. Depending on the type of secretion, type of particle, BMBF-funded research consortia OPTOPROBE and GLUTEVIS: its size and surface properties the particles behave differently ATTO-TEC, Siegen; University of Applied Sciences, Paderborn; in the mucosal environment. They either agglomerate in sec- GeSiM, Dresden; Karl Storz, Tuttlingen; LMTB, Berlin; R-Bio- retions, stick to cells and/or mucin or are excreted along with pharm, Darmstadt; DFA, Freising; Hermann Kröner Stärke, fecal matter. Yet, substantial translocation rates into the body Ibbenbüren could not be observed. In light of this it seems questionable SPP Biomedizintechnik: University of Lübeck whether nanoparticulate matter can significantly contribute to DZL German Center for Lung Research: LMU München allergic sensitization at mucosal surfaces by acting as adjuvant or allergen carrier. GRANT SUPPORT Adaptive immune responses as they occur e.g. in allergic sen- Bundesministerium für Bildung und Forschung (BMBF), grants sitizations are distinctive and complex events. They depend, 13N10505 & 13GW0042 amongst others, on the B- and T-cell repertoire at the time of Deutsche Forschungsgemeinschaft (DFG), grant Fr 958/5-2 allergen contact, the physical integrity of the allergen, the site University of Lübeck, grant F249607 of contact, and on the actual status of cytokines and other German Center for Lung Research (BMBF) AA2.2 & COPD-2 humoral mediators. Consequently, the recognition of allergen by the immunoglobulins induced may differ from individual to individual in terms of affinity, epitopes bound and antibody class. In order to shed light on this complex response, we have established a high-throughput epitope recognition assay based on synthetic peptide libraries and are using these libraries for epitope mapping of allergens in allergic disorders. SELECTED PUBLICATIONS Sinnecker H, Krause T, Koelling S, Lautenschläger I, Frey A. The gut wall provides an effective barrier against nanoparticle uptake. Beilstein J Nanotechnol 2014, 5:2092-2101. STRUCTURAL BIOCHEMISTRY • PROF. DR. OTTO HOLST LIPOPHILIC ALLERGENS ENDOTOXIN-FREE ESCHERICHIA COLI ANTIBODY/LECTIN BINDING TO MICROBIAL SURFACES MISSION STRUCTURAL ANALYSIS OF HIGHLY PURIFIED, BIOLOGICALLY RELEVANT, BACTERIAL AND ENVIRONMENTAL COMPOUNDS AIMING IN UNDERSTANDING THEIR ROLE IN THE COURSE OF ALLERGIC INFLAMMATION. PATHOMECHANISMS OF STENOTROPHOMONAS MALTOPHILIA. STRUCTURAL AND FUNCTIONAL ANALYSES OF ANTIBODIES AND LECTIN BINDING TO MICROBIAL SURFACE GLYCANS. IDENTIFICATION OF CARBOHYDRATE EPITOPES TO BE USED FOR DIAGNOSTIC AND THERAPEUTIC PURPOSES AND TO REVEAL BINDING MECHANISMS. MOST IMPORTANT FINDINGS Lipid compounds originating from HDM and mite feces were During allergic asthma, an IgE-mediated immune response is fractionated and characterized by HPTLC, GC and GC/MS. A triggered against proteins like pollen and house dust mite all- broad spectrum of lipid-containing molecules, with qualitative ergens. However, peptides and glycopeptides are not the sole and quantitative differences between both samples, was ob- agents present in pollen grains or mite feces which contain also served. a wide range of (glyco-)lipids that can be recognized by NKT cells Recombinant proteins manufactured in Escherichia coli are via CD1d receptor, contributing to the allergic response, thus inherently contaminated with constituents of the bacterial cell acting as adjuvants triggering innate signaling pathways. Inte- such as LPS, a potent agonist for hTLR4/MD-2-mediated pro- restingly, it is known that several major allergens of different inflammatory activity in immune cells. To prevent such toxicity, sources have hydrophobic domains able to interact with lipids. we have constructed a series of LPS-lacking E. coli strains which In order to test whether (glyco-)lipids can act as ajuvants contri- are viable despite exclusively elaborating the LPS precursor li- buting to the development of the allergenic response to aeroall- pid IVA, and cannot easily regain the ability to synthesize toxic ergens, timothy grass grains (Phleum pratense), and house dust LPS (collaboration with Research Corporation Technologies Inc., mite (HDM) (Dermatophagoides pteronyssinus) and their feces Tucson, AZ, and the Dept. of Medicinal Chemistry, University of were extracted and fractionated. Pure ceramides, phytoceramides Michigan, Ann Arbor, MI). The mutant cells can be used to pro- and diglycolyl diacylglycerol were isolated from pollen grains (Fig. 1), duce recombinant proteins that are free of endotoxic activity in and their structures were determined by GC/MS, ESI MS and NMR human LPS-responsive cells, thus offering a wide range of ap- spectroscopy. Two fractions containing ceramides, phytoceramides plications in biotechnology, molecular biology and cell biology. and diglycolyl diacylglycerol activated iNKT cells in vitro in a CD1d- They can also be used to manufacture functional endotoxin-free restricted manner, and intranasal administration of pollen-derived inclusion bodies as promising self-organizing catalytic materials glycolipids induced a type 2 response in a mouse model. in drug delivery and regenerative medicine. The strains are now commercially available from RCT and Lucigen Corp., Middleton, WI, under the registered trademark ClearColi® (Nature Methods: http://www.nature.com/nmeth/journal/v10/n9/abs/ nmeth.f.367.html). The ClearColi® expression technology has been awarded as one of the „Top 10 Innovations of 2013“ by The Scientist magazine (http://www.the-scientist.com/?articles. view/articleNo/ 38394/title/Top-10-Innovations-2013/). In collaboration with S. Evans (University of Victoria, Canada) and P. Kosma (University of Natural Resources and Life Sciences, Vienna, Austria) we resolved the structure of the monoclonal antibody (mAb) S25-26 in complex with its high-affinity ligand from LPS of Chlamydia. Importantly, this mAb was glyco Figure 1: Purification scheme of lipophilic compound of Phleum pratense, with indication of obtained pure compounds. sylated in the variable domain of the heavy chain. Analysis of the glycans revealed a heterogeneous mixture with an unusually Seite 67 / PRA Asthma and Allergy UTE AGGE • HELGA BARTELS • RAINER BARTELS • PETRA BEHRENS • KATARZYNA DUDA • SYLVIA DÜPOW • REGINA ENGEL • DÖRTE GRELLA • VOLKER GROTE • MANUEL HAIN • OTTO HOLST • KATHARINA JAKOB • GUDRUN LEHWARKYVETOT • UWE MAMAT • SVEN MÜLLER-LOENNIES • SANDRA PETERSEN • WIEBKE SCHNOOR • VERONIKA SUSOTT high number of terminal αGal-Gal moieties (αGal-epitope), like schel A, Xia G. Wall teichoic acid structure governs horizontal the therapeutic Ab Cetuximab. The elucidation of the three-di- gene transfer between major bacterial pathogens. NATURE mensional structure of an αGal-containing N-linked glycan has COMMUNICATIONS. 2013; 4:2345. potential clinical interest, as it has been implicated in allergic re- Silipo A, Vitiello G, Gully D, Sturiale L, Chaintreuil L, Fardoux sponse in patients receiving therapeutic antibodies. One of the J, Gargani D, Lee H-I, Kulkarni G, Busset G, Marchetti R, Pal- unliganded structures of S25-26 for the first time gave insight migiano A, Moll H, Engel R, Lanzetta R, Paduano L, Parrilli into the 3-dimensional structure of this important allergenic car- M, Chang W-S, Holst O, Newman DK, Garozzo D, D‘Errico G, bohydrate antigen. Giraud E, Molinaro A. Covalently linked hopanoid-lipid A im- Also, we obtained the single-chain variable domain (scFv) proves outer-membrane resistance of a Bradyrhizobium sym- M6P-1, a unique Ab fragment with specificity for mannose- biont of legumes. NATURE COMMUNICATIONS. 2014; 5:5106. 6-phosphate (Man6P) monosaccharide. The acquisition of Raedler D, Ballenberger N, KluckerE, Böck A, Otto R, Pra- Man6P on N-linked glycans of enzymes is a structural requi- zeres da Costa O, Holst O, Illig T, Buch T, von Mutius E, rement for their transport from the Golgi apparatus to lysoso- Schaub B. Identification of novel immune phenotypes for mes. We were able to show that this scFv binds mono- and allergic and non-allergic childhood asthma. JOURNAL OF AL- diphosphorylated Man6 and Man7 but not phosphodiesters, LERGY AND CLINICAL IMMUNOLOLOGY. 2014; doi: 10.1016/j. monophosphorylated Man8 or mono- or diphosphorylated jaci.2014.07.046. Man9 structures. Single crystal X-ray diffraction analysis of Fv Komaniecka I, Choma A, Mazur A, Duda KA, Lindner B, M6P-1 in complex with Man6P revealed that specificity and af- Schwudke D, Holst O. Occurrence of an unusual hopanoid- finity is achieved via multiple hydrogen bonds to the mannose containing lipid A among lipopolysaccharides from Bradyrhi- ring and two salt-bridges to the phosphate moiety. Loss of bin- zobium species. THE JOURNAL OF BIOLOGICAL CHEMISTRY. ding was observed for scFv M6P-1 at pH values below the se- 2014; pii: jbc.M114.614529. [Epub ahead of print]. cond pKa of Man6P (pKa 6.1). The structures of Fv M6P-1 and Rueda F, Cano-Garrido O, Mamat U, Wilke K, Seras-Franzo- the cellular Man6P-receptors suggest that the change of the so J, Garcia-Fruitos E, Villaverde A. Production of functional ionization state of Man6P is the main driving force for the loss inclusion bodies in endotoxin-free Escherichia coli. APPLIED of binding at acidic pH (e.g. lysosome pH 4.8), which provides MICROBIOLOGY AND BIOTECHNOLOGY. 2014; 98:9229-9238. justification for the evolution of a lysosomal enzyme transport Haji-Ghassemi O, Müller-Loennies S, Saldova R, Muniyappa pathway based on Man6P recognition. M, Brade L, Rudd PM, Harvey DJ, Kosma P, Brade H, Evans Figure 2: Crystal SV. Groove-type recognition of chlamydiaceae-specific lipopo- structure of lysaccharide antigen by a family of antibodies possessing an mannose-6-phospha- unusual variable heavy chain N-linked glycan. THE JOURNAL te bound to scFv M6P-1 and view of a monophosphorylated Man6N-glycan (right top). OF BIOLOGICAL CHEMISTRY. 2014; 289:16644-16661. Madhavarao CN, Agarabi CD, Wong L, Müller-Loennies S, Braulke T, Khan M, Anderson H, Johnson GR. Evaluation of butyrate-induced production of a mannose-6-phosphorylated therapeutic enzyme using parallel bioreactors. BIOTECHNO- PUBLICATIONS LOGY AND APPLIED BIOCHEMISTRY. 2014; 61:184-192. Semeniuk A, Sohlenkamp C, Duda K, Hölzl G. A bifunctional glycosyltransferase from Agrobacterium tumefaciens synthe- INTERNAL AND EXTERNAL COLLABORATIONS sizes monoglucosyl and glucuronosyl diacylglycerol under 13 INTERNAL AND 31 EXTERNAL COLLABORATIONS phosphate deprivation. THE JOURNAL OF BIOLOGICAL CHEMISTRY. 2014; 289:10104-10114. GRANT SUPPORT Winstel V, Liang C, Sanchez-Carballo P, Steglich M, Munar M, SFB TR 22 (DFG); DZL; EU COST “Cystic Fibrosis”; VW Stiftung Bröker BM, Penadés JR, Nübel U, Holst O, Dandekar T, Pe- CENTER FOR CLINICAL STUDIES • DR. CHRISTIAN HERZMANN CLINICAL STUDIES TRANSLATIONAL RESEARCH TUBERCULOSIS ASTHMA MISSION THE CENTER FOR CLINICAL STUDIES IS AT THE INTERFACE BETWEEN PATIENT CARE AND BASIC RESEARCH. WE PROVIDE SCIENTISTS WITH ACCESS TO PATIENTS AND CLINICAL SPECIMENS. WE EMPOWER CLINICIANS TO INVESTIGATE THE DISEASES THEY FACE EVERY DAY. WE MAINTAIN AND IMPROVE THE INFRASTRUCTURE THAT IS REQUIRED FOR INVESTIGATOR INITIATED AND SPONSOR INITIATED TRANSLATIONAL RESEARCH. DEVELOPMENT asthma exacerbation. An investigator initiated scientific add- In 2014, we optimized our infrastructure through harmoni- on to this project is currently in the pipeline in collaboration zation of our own SOPs with the requirements of the Clinical with the research group Asthma Mouse Models. Trials Unit of the German Centre for Infection Research (DZIF). For the TB or not TB consortium (www.tbornottb.de), patient Furthermore, the TB or not TB consortium ended and created recruitment (n=20) continued while parallel data analyses a vast amount of unique data on host resistance to tubercu- were performed. The analysis suffered from the retirement of losis. This affected the head of our unit directly, as he guided Dr. Martin Ernst (former research group of Immune Cell Biolo- the development of the project for the past five years. For the gy) who performed many of the experiments. In collaboration first time since the foundation of our unit we were able to es- with Dr. G. Sotgiu (Italy) we confirmed that early childhood vac- tablish and maintain the infrastructure for phase I studies. cination with M. bovis BCG is not associated with protection against M. tuberculosis infection in an ex vivo growth inhibi- MOST IMPORTANT PROJECTS tion assay (published in the European Respiratory Journal). Further analyses on the cytokine profiles of individual patients in response to mycobacterial infections are currently ongoing in collaboration with the research groups Microbial Interface Biology and Bioinformatics. Aiming to improve the diagnostic options for children with tuberculosis, a sorbent chewing gum for the detection of M. tuberculosis in the oral cavity is currently under development in collaboration with the research groups Microbial Interface Biology, Figure 1: Geographic distribution of ExplainTB users in 2014. The project has been accessed from more than 80 countries (light blue), most frequently from Germany (dark blue). Structural Biochemistry and the National Reference Labo- The very first phase I study at the Research Center Borstel ratory for Micobacteria. The initial phase of the TuberChewLo- was conducted in collaboration with Janssen on an investiga- sis project was funded by the VolkswagenFoundation and ser- tional asthma drug (Clinical trials identifier NCT01704040). ved the identification of potential capture mechanisms. Following an intensive preparation period of 18 months, we We further expanded the multilingual tuberculosis education were able to recruit patients a multicentre study investigating project ExplainTB (www.explaintb.org). Currently, ExplainTB of- a TLR-9 antagonist under investigation for the prevention of fers freely available educational material for tuberculosis pa- Seite 69 / Medicine JOHANNA DÖHLING • ANDREA GLAEWE • LENKA KRABBE tients and relatives in 38 languages. Users access the project thorities Hanover/Frankfurt – Institute for Lung Research – website from more than 80 countries worldwide. The initiative Koch-Metschnikoff-Forum – Max-Planck-Institut for Infection was covered by several national newspapers (e.g. Süddeut- Biology – Tb or not Tb Consortium – B-NET – University Hos- sche Zeitung, ZEIT, Frankfurter Allgemeine Sonntagszeitung) pital of Ulm, Institute for Hygiene and Medical Microbiology and was awarded the Aspirin Social Award 2014 by the Bayer – University of Cork, Ireland, School of Food and Nutritional Cares Foundation. Furthermore, it was Top-6-shortlisted for Sciences the European Health Award by the European Health Forum Internal collaborations Gastein. While the project was co-ordinated at the Research Biophysics – Bioinformatics – Clinical Infectious Diseases – Center Borstel, the contributions were made by more than Immunobiophysics – Immunochemistry – Microbial Interface 400 volunteers worldwide whom we are deeply indebted to. Biology – BioMaterialBank North Figure 2: Chewing gum base Artica T® used for the TuberChewLosis project that aims to develop a new diagnostic method for the detection of M. tuberculosis in the oral cavity. SELECTED PUBLICATIONS Herzmann C, Sotgiu G, Schaberg T, et al. Early BCG vaccination is unrelated to pulmonary immunity against Mycobacterium tuberculosis in adults. Eur Respir J. 2014 Oct;44(4):1087-90 Thoden J, Potthoff A, Bogner JR, Brockmeyer NH, Esser S, Grabmeier-Pfistershammer K, Haas B, Hahn K, Härter G, Hartmann M, Herzmann C, et al. Therapy and prophylaxis of opportunistic infections in HIV-infected patients: a guideline by the German and Austrian AIDS societies (DAIG/ÖAG) (AWMF 055/066). Infection. 2013 Sep;41 Suppl 2:S91-115 INTERNAL AND EXTERNAL COLLABORATIONS Industry partners: AID – Alere Technologies – AstraZeneca – CAFOSA – Janssen – Mologen – Mundipharma – Roche – PulmonX – Revolymer – Siemens Academic partners: Bernhard-Nocht-Institute – CAP-Netz Foundation – Health au- CLIN. & EXP. PATHOLOGY • PROF. DR. MED. DR. MED. VET. DR. H.C. EKKEHARD VOLLMER HUMAN EX VIVO MODEL HOPE-TECHNIQUE TRANSCRIPTOME / PROTEOME / METHYLOME MISSION EXCELLENT DIAGNOSTICS AND RESEARCH IN PNEUMOLOGY. CENTRAL EXPERIMENTAL TOOLS REMAIN FUNCTIONAL HUMAN TISSUE CULTURE MODELS FOR DIFFERENT DISEASES OF THE LUNG, MOSTLY BASED ON THE HOPE-TECHNIQUE. EXPERIMENTAL WORK ON PRIMARY HUMAN LUNG CELLS IS A FURTHER IMPORTANT TOOL. FOR SCREENING PURPOSES AND BIOMARKER IDENTIFICATION WE APPLY INHOUSE TRANSCRIPTOME ANALYSES, A BUNCH OF MODERN MOLECULAR TECHNIQUES SERVE FOR VALIDATION. MOST IMPORTANT FINDINGS Using the human ex vivo tissue culture model, we have We continued the work on BAMBI (Bone morphogenetic pro- shown complex interactions between Legionella pneumo- tein and Activin Membrane-Bound Inhibitor), which was disco- phila and the human host in cooperation with the University vered in the human lung by our group, within the German Cen- Braunschweig. ter for Lung Research (DZL) in cooperation with the Human Together with the LungenClinic Grosshansdorf we developed Genetics at the University of Kiel and the Systems Biology at a procedure which allows for HOPE-preservation and paraffin- the German Cancer Center (DKFZ). embedding of induced sputum samples (Figure 1). Figure 1: Exemplary heat map of transcriptome analysis of Figure 2: Hierarchical cluster analysis and heatmap of 109 CpG loci HOPE-preserved IS RNA processed with and without DTT. Agilent found differentially methylated in cryo-preserved, HOPE- fixed and/or 4x44k human whole genome arrays have been used. Analysis of the formalin-fixed sample tissues as determined by an ANOVA results was performed with a fold change analysis using GeneSpring (FDR<0.05; σ/σ max>0.2). Except cryo-preserved tissues all samples 12 software. Red color intensity indicates higher expression, green were paraffinized. Only data obtained from tumor free lung tissues color intensity indicates lower expression, and white color indicates were included. Blue bars: cryo-preserved samples, green bars: a medium level of expression of a gene in the comparison of both HOPE-fixed and pink: formalin-fixed tissue samples. Heatmap: blue: groups. Gene description is shown on the left hand side. (from low, yellow: high DNA methylation values (from Marwitz et al., Lab Goldmann et al., Respiration 2013;86(3):262-3). Invest. 2014 Aug;94(8):927-33). In cooperation with the Univ. Kiel and the Helmholtz-Center in The DZL laboratory for primary cells and ex vivo models is run Braunschweig we have shown that HOPE-fixed, paraffin em- together with the Experimental Pneumology Borstel and the bedded tissues can be used for methlylome and proteome LungenClinic Grosshansdorf, a facility which provides primary analyses including phosphoproteomes (Figure 2). human lung cells and two ex vivo systems based on human Seite 71 / Medicine BETTINA BARON-LÜHR • GABRIELE CORNEHLS • STEFFI FOX • DR. DARIIMAA GANBAT • PD DR. KAROLINE GAEDE • PROF. DR. TORSTEN GOLDMANN • LYDIA GUSE • IRIS JONAS • ALINA KELP • ANGELA KELP • BRITTA KRÖGER • DR. DAGMAR S. LANG • MARIA LAMMERS • MILENA LEMBURG • CARMEN LICHT • BELINDA LIE • DR. SEBASTIAN MARWITZ • JULIA MÜLLER MD • DR. FRAUKE PEDERSEN • ROMINA PRITZKOW • ANNA SCHILLER • DR. BERNHARD SCHMITT • DR. SOPHIE SEEHASE • DR. FLORIAN STELLMACHER • JASMIN TIEBACH • ROLF WARNECKE • DR. IRIS WATERMANN • KRISTIN WICZKOWSKI tissues, precision cut lung slices (PCLS) and short term stimu- Greinert, B. Karlsdorf), Inflammation and Regenaration (H. lation of tissues (STST). Fehrenbach, Ch. Vock),Barrier Integrity (I. Lautenschläger), In the DZL we are engaged in multiple research projects con- Structural Biochemistry (O. Holst), Biophysics (T. Gutsmann, cerning COPD, asthma, and lung cancer. K. Brandenburg), Biochemical Immunology (F. Petersen), Cel- The Clin. & Exp. Pathology is responsible for many biobanking lular Pneumology (C. Stamme), Innate Immunity (H. Heine), activities (Biomaterialbank Nord) covering diverse projects in Fluorescence Cytometry (T. Scholzen, J. Behrens), Bioanalyti- the DZL and others. cal Chemistry (D. Schwudke) Furthermore, in close cooperation with the National Refe- External collaborations: Airway Research Center North (ARCN) rence Center for Mycobacteria(NRZ), we expanded the range in the German Center for Lung research (DZL). University of of molecular diagnostics in the lung by implementation of Lübeck, Med Clinic III (P. Zabel, K. Dalhoff, D. Drömann). Uni- NRAS-mutations, which are decision-making in targeted the- versity of Lübeck, Medical Microbiology (J. Rupp). Lungen- rapies of lung cancer. Clinic Grosshansdorf (Rabe K, Reck M, H. Watz, Ch. Kugler, F. Pedersen, L. Welker). University of Kiel (O. Ammerpohl, R. Siebert), University of Dohuk, Iraq (M. Abdullah). Tblisi Cancer SELECTED PUBLICATIONS Center, Georgia (A. Mariamidze). Yerevan State Medical Uni- Marwitz S, Kolarova J, Reck M, Reinmuth N, Kugler C, Schäd- versity, Armenia (A. Mkhitarian). University of Freiburg, Pneu- lich I, Haake A, Zabel P, Vollmer E, Siebert R, Goldmann T, mology (J. Müller-Quernheim, G. Zissel). BMBF Medical Infec- Ammerpohl O. The tissue is the issue: improved methylome tion Genomics: University of Braunschweig, Helmholtz-Center analysis from paraffin-embedded tissues by application of the Braunschweig, Robert Koch Institute Wernigerode University HOPE technique. Lab Invest. 2014 Aug;94(8):927-33 of Greifswald (M. Steinert, A. Flieger, M. Hecker, L. Jänsch). Shevchuk O, Abidi N, Klawonn F, Wissing J, Nimtz M, Kugler GRANT SUPPORT C, Steinert M, Goldmann T, Jänsch L. HOPE-fixation of lung tis- BMBF: DZL Airway Research Center North: Funding for pro- sue allows retrospective proteome and phosphoproteome stu- jects in the areas of COPD, Lung Cancer, Asthma and Bioban- dies. Journal of Proteome Research 2014 7;13(11):5230-9. king. BMBF project on the ex vivo infection of human lung tissues with Legionella pneumophila (Legioprotect). Jäger J, Marwitz S, Tiefenau J, Rasch J, Shevchuk O, Kugler C, Goldmann T, Steinert M. Human lung tissue explants reveal DFG grant in cooperation with D. Drömann, University of Lü- novel interactions during Legionella pneumophila infections. beck, Med. Clinic III: Charakterisierung TGF-ß inhibierender Infect Immun. 2014 Jan;82(1):275-85. Moleküle bei inflammationsgetriggerten Remodelingprozessen der Lunge Goldmann T, Pedersen F., Seehase S, Marwitz M, Lang DS, Kirsten AM, Zabel P, Vollmer E, Magnussen H, Rabe KF, Watz H. The effect of dithiothreitol on the transcriptome of induced sputum cells. Respiration 2013;86(3):262-3. INTERNAL AND EXTERNAL COLLABORATIONS Internal collaborations: Medical Clinic (P. Zabel), NRZ(S. RüschGerdes, E. Richter, D. Hillemann), Molecular Mycobacteriology (S. Niemann), Microbiology and Infectiology (U. Schaible), Microbial Inflammation Research (S. Ehlers, N. Reiling), Clinical and Molecular Allergology (U. Jappe), Mucosa Immunology (A. Frey, K. Ramaker), Clinical Infectious Diseases (C. Lange, U. NATIONAL REFERENCE CENTER FOR MYCOBACTERIA • DR. SABINE RÜSCH-GERDES TUBERCULOSIS NONTUBERCULOUS MYCOBACTERIA RAPID DETECTION OF MULTIDRUG AND EXTENSIVELY DRUG RESISTANCE (MDR AND XDR) LAB STRENGTHENING EXTERNAL QUALITY ASSURANCE MISSION AS PART OF THE WORLDWIDE LABORATORY NETWORK OF THE WHO, THE GERMAN NATIONAL REFERENCE LABORATORY FOR MYCOBACTERIA (NRL) ACTS AS A SUPRANATIONAL REFERENCE LABORATORY SINCE 1995. SINCE 2008, THE NRL COORDINATES AND PROVIDES SAMPLES FOR EXTERNAL QUALITY ASSURANCE IN THE FIELD OF MYCOBACTERIAL DIAGNOSTICS, ORGANIZED BY INSTAND E-V. DÜSSELDORF. ON A DAILY BASIS CONSULTANCY IS PROVIDED FOR LABORATORIES AND CLINICS CONCERNING DIAGNOSTICS AND THERAPY OF MYCOBACTERIAL DISEASES. THE RESEARCH OF THE NRL FOCUSES ON THE TOPICS “EVALUATION OF NEW TECHNIQUES FOR THE IMPROVEMENT OF DIAGNOSTICS”, “RESISTANCE OF TUBERCULOSIS”, AND “VARIABILITY OF CLINICAL MYCOBACTERIAL ISOLATES”. MOST IMPORTANT FINDINGS evaluate the RMP susceptibility with different DST methods. Primarily the NRL is a routine laboratory specialized to isola- In total 143 INH resistant/RMP susceptible M. tuberculosis te mycobacteria, identify mycobacterial species with classical isolates submitted to the NRL in 2011 were analyzed. Sequen- and molecular methods, and perform susceptibility testing for cing of the rpoB-RRDR revealed no mutation in 139 of the all available drugs. In recent years the global situation con- 143 strains while four strains (2.8%) had at least one rpoB mutation in the RRDR. Two of the four strains had the mutation L533P. The other two strains had a mutation D516Y in combination with a second mutation within the RRDR (N518D and E510H). Analysis of the RMP MIC values below the standard critical concentrations revealed different MICs for the four strains. Three of the four strains showed slightly elevated MICs but below the critical concentration while one strain showed no elevated MIC when performing DST on BACTEC MGIT 960 or the proportion method on LJ. We conclude from these data that both methods are equally reliable for phenotypic DST and MIC determination. Figure 1: RMP MICs below the critical concentrations (---) determined by Bactec 960 MGIT and the proportion method on LJ cerning drug resistance has changed dramatically. In order to combat this, the NRL focused to research on the development of resistance and epidemiology of drug resistant TB, combining advanced microbiological and molecular methods. Isoniazid (INH) resistant/RMP susceptible strains represent the main source for the development of multidrug resistant strains. Mycobacterium tuberculosis isolates that harbor rpoB mutations within the rifampicin (RMP) resistant determining region (RRDR) and test RMP susceptible by phenotypical drug susceptibility testing (DST) methods limit the reliability of the results obtained from molecular assays. We investigated the presence of rpoB mutations in phenotypically INH resistant/ RMP susceptible strains that were isolated from patients in Germany to analyze the frequency of this constellation and to Seite 73 / Medicine DR. SÖNKE ANDRES • KRISTINE BEUK • MANUELA DORN • GUDRUN HEINONEN • DR. DORIS HILLEMANN • SILVIA HÖLLGER • MARGRIT KERNBACH • JANINA KOLB • ANNE-KATHRIN LANDGRAF • KIRSTEN OTT • ILSE RADZIO • PD DR. ELVIRA RICHTER • FRAUKE SCHAEFER • BIRTE SCHLÜTER • DANIELA SIEVERT • PETRA VOCK • BIRGIT VOSS • ANNKATHRIN WITT Figure 2 & 3: Occult rpoB mutations produce „resistant“ genotypes by the frequently used GeneXpert (Cepheid) and MTBDR-plus (HAIN) sceptible Mycobacterium tuberculosis isolates from Germany. Antimicrob Agents Chemother. 2014, 58(1): 590-592 line-probe assay Recent studies have do- Kolb J, Hillemann D, Möbius P, Reetz J, Lahiri A, Lewin A, cumented that nontuber- Rüsch-Gerdes S, Richter E. Genetic characterization of Ger- culous mycobacteria have man Mycobacterium avium strains isolated from different outnumbered tuberculosis hosts and specimens by multilocus sequence typing. Int J in some aereas. Among Med Microbiol. 2014, S1438-4221 mycobacterial opportuni- stic pathogens, M. avium Hillemann D, Hoffner S, Cirillo D, Drobniewski F, Richter hominissuis is the most E, et al. First Evaluation after Implementation of a Quality important for humans. In Control System for the Second Line Drug Susceptibility Tes- contrary to tuberculosis, ting of Mycobacterium tuberculosis Joint Efforts in Low and research on the characte- High Incidence Countries. PLoS ONE 2013, 8(10): e76765. rization of specific strains doi:10.1371 and their association with human infection is in the INTERNAL AND EXTERNAL COLLABORATIONS beginnings. The assump- International collaborations have been established with seve- tion environmental ral countries e.g. Austria, Armenia, Azerbaijan, Bosnia-Herze- strains and not a subgroup of highly virulent strains are the gowina, Croatia, Ghana, Kazakhstan, Moldova, Serbia, Sierra causative agents is not proven yet. To investigate the strain Leone, Slowenia, and Uzbekistan. The NRL consults UNDP, diversity, reliable and standardizable typing methods were es- MSF, WHO, diagnostic laboratories/health authorities, trains tablished: Multilocus sequence typing (MLST) and mycobac- more than 80 persons per year, and cooperates with national terial interspaced repetitive units-variable number tandem (RKI, Robert Koch Institute; public health offices) and inter- repeats (MIRU-VNTR). Both methods were applied to charac- national centers/organizations (WHO; World Health Organiza- terize a set of M. avium hominissuis strain isolated from hu- tion; USAID, U.S. Agency for the International Development; mans (children and adults) and from slaughter pigs. FIND; The Foundation for Innovative New Diagnostics). Interestingly, it could be shown that several allelic profiles As part of the WHO/Union Global Project for Surveillance of were present in both human and pig isolates, which clearly TB Drug Resistance the NRL annually participates in rounds indicates that the infections with M. avium strains are caused of proficiency testing to assure quality of drug susceptibili- by environmental strains and not specifically by highly virulent ty testing for all partner laboratories. To improve the quality clones. In addition, new strain profiles were found. The high standard of diagnostic laboratories in Europe the NRL was ap- degree of diversity within ‚M. avium subsp. hominissuis‘ as pointed member of the „Management Team“ by the European well as the relatedness of human, porcine and environmen- Centre for Disease Prevention and Control (ECDC) program tal strains could be confirmed by IS1245 RFLP fingerprinting. to improve TB-diagnostic in Europe. Since 2010 samples for Furthermore, it could be shown that the PCR for detection of quality control for 31 countries were produced, sent, and the IS901 is not a definitive proof of M. avium subsp. avium/M. incoming data analysed. that avium subsp. silvaticum, because it was detected in a strain, displaying ‚M. avium subsp. hominissuis‘ MLST profile. GRANT SUPPORT BMG, DZK-SH, ECDC, EU TB PAN-NET, EU-Nareb, INSTAND, SELECTED PUBLICATIONS RKI (National Reference Laboratory and network respiratory Andres S, Hillemann D, Rüsch-Gerdes S, Richter E. Occur- infections), WHO rence of rpoB mutations in isoniazid-resistant but rifampin-su- FLUORESCENCE CYTOMETRY • DR. J. BEHRENDS/DR. T. SCHOLZEN FLOW CYTOMETRY CELL SORTING CONFOCAL LASER SCANNING MICROSCOPY LIVE CELL IMAGING BIOSAFETY LEVEL 3 SUPPORT/SERVICE/TEACHING MISSION THE RESEARCH STRATEGY OF THE FLUORESCENCE CYTOMETRY BORSTEL (FCB) IS FOCUSED ON THE COMPREHENSION OF MOLECULAR AND CELLULAR INTERACTIONS IN THE FIELDS OF INFLAMMATION, INFECTION AND ALLERGY. THE ANALYSIS OF SUCH PROCESSES REQUIRES THE ABILITY TO PHENOTYPICALLY CHARACTERIZE INDIVIDUAL CELLS (FIG. 1) CONCERNING THE EXPRESSION OF SURFACE AND INTRACELLULAR MOLECULES (E.G. RECEPTORS OR TRANSCRIPTION FACTORS) OR SECRETED COMPONENTS (E.G. CYTOKINES). THESE REQUIREMENTS ARE WELL MET BY FLOW CYTOMETRY, WHICH IS SUITED TO ANALYZE LARGE NUMBERS OF CELLS IN A QUANTITATIVE MANNER. MOREOVER, IT IS POSSIBLE TO PURIFY INDIVIDUAL POPULATIONS OF LIVING CELLS BY FACS FOR SUBSEQUENT EXPERIMENTS. HOWEVER, TO GET ADDITIONAL INFORMATION ABOUT THE SPATIAL DISTRIBUTION OF COMPONENTS AND THE CHRONOLOGICAL SEQUENCE OF EVENTS, HIGH RESOLUTION LIVE CELL IMAGING IS A KEY TECHNIQUE FOR THE ANALYSIS OF COMPLEX PROCESSES. THE SERVICE UNIT FCB OFFERS SCIENTISTS AT THE RCB AND ASSOCIATED UNIVERSITIES ACCESS TO MODERN FLUORESCENCE-BASED TECHNIQUES BY PROVIDING BOTH, STATE-OF-THE-ART FLOW CYTOMETERS AND FLUORESCENCE MICROSCOPES (FIG. 2). IN OCTOBER 2013 THE STAFF HAS BEEN INCREASED AND CONSISTS NOW OF ONE TECHNICIAN AND TWO SCIENTISTS. DUE TO THIS IMPROVEMENT, THE SCOPE OF SERVICE AND TRAINING COULD BE EXPANDED, SO THAT SCIENTISTS AT THE RCB ARE NOW OFFERED A COMPREHENSIVE THEORETICAL AND PRACTICAL SUPPORT WITHIN THE FIELD OF CYTOMETRY. MOST IMPORTANT FEATURES sers (blue and red) and detecting up to 4 colors. High speed FLUORESCENCE MICROSCOPY: High resolution (live cell) ima- cell sorting is available on our BD FACSAriaII flow cytometer. ging is a key technique to obtain information about the spatial In December 2014 this sorter was upgraded with new fluidic distribution of components and the chronological sequence systems, sorting chamber, computer and FACSDiva™ 8.0. of events. The Olympus IX-81 is a live cell imaging system, including an incubation chamber with CO2 gassing as well as temperature and humidity regulation. With 4 fluorescence channels, a highly sensitive CCD camera and a motorized stage it is well-suited for time lapse imaging. The Leica TCS SP5 is an inverse confocal laser scanning microscope equipped with 5 fluorescence detectors. In November 2014 the system was upgraded with two Hybrid detectors, increasing sensitivi- Figure 1: Flow cytometry is suited to analyze large numbers of cells in a quantitative manner and in contrast Fluorescence Microscopy is ty, permitting reduced photodamage in live cell experiments. a key technique for the spatial-temporal analysis of complex The microscope contains an incubation chamber with tempe- processes. rature regulation and CO2 gassing. The system can be used This upgrade leads to increased operational reliability, easier for conventional image analysis as well as live cell imaging handling and thus doubling the available sort time per day. including FRET and FRAP experiments. With 3 lasers (violet, blue and red), analysis of up to 9 fluo- FLOW CYTOMETRY: Flow cytometry allows to analyze large rescence channels is possible, 4 populations can be sorted numbers of cells in a quantitative manner. The BD LSR II simultaneously. The analyzers BD FACSCantoII (3 lasers, 8 flow cytometer is especially well equipped for multiparameter colors detection) and BD FACSArray (2 lasers, 4 colors detec- analysis, includes 3 lasers (ultraviolet, blue, red) and a high tion) are housed in a Biosafety Level 3 facility within biosafety throughput sampler that can rapidly introduce samples from cabinets, allowing analysis of biohazardous material. 96 and 384 well plates. The analysis of up to 8 fluorescence TRAINING: Technical training is an integral part of the FCB channels is possible, including the measurement of calcium concept. Basic training is provided for all instruments of the influx using an UV laser. Additionally analysis can be perfor- FCB. Additionally, advanced training is offered on an indivi- med using a BD FACSCalibur cytometer, equipped with 2 la- dual basis depending on the particular project. In 2014, 35 Seite 75 / Central Units MARTINA HEIN SERVICE: The service unit FCB offers scientists to carry out service for high speed cell sorting on our BD FACSAriaII flow cytometer. The service unit is also available for consultation about the design of individual experiments, the analysis of data (Fig. 3) and can also take on joint research projects. SELECTED METHODS: - -multiparameter analysis of cell populations - cell proliferation/viability assays - FRET/FRAP, co-localization analysis - sorting of e.g. DCs, B cells, T cells, cell lines - metabolic analysis of different Mtb strains Figure 2: The Core Facility FCB provides access to state-of-the-art SELECTED PUBLICATIONS flow cytometers, sorter and confocal microscopes as well as a live Heitmann L, Abad Dar M, Schreiber T, Erdmann H, Behrends cell imaging system. J, McKenzie A.N., Brombacher F, Ehlers S and Hölscher C. The scientists and technicians received a basic training for one IL-13/IL-4R-alpha axis is involved in tuberculosis-associated pa- of the instruments. Supporting theoretical training courses thology. JOURNAL OF PATHOLOGY 2014 Nov;234(3):338-50. are also offered. In 2013/2014 users could choose from 18 different seminars covering various topics from the fields of Schneider BE, Behrends J, Hagens K, Harmel N, Shayman JA, flow cytometry and fluorescence microscopy. The FCB is also and Schaible UE. Lysosomal phospholipase A : A novel player committed to the training of young academics and takes an in host immunity to Mycobacterium tuberculosis. EUROPEAN active part in practicals for students from the Universities of JOURNAL OF IMMUNOLOGY 2014 Aug;44(8):2394-404. Lübeck and Kiel. The core facility is also integrated into the Borstel Biomedical Research School for graduates. Everts B, Hussaarts L, Driessen NN, Meevissen MH, Schramm, G, van der Ham AJ, van der Hoeven B, Scholzen T, Burgdorf, S, Mohrs M, Pearce EJ, Hokke CH, Haas H , Smits HH, Yazdanbakhsh M. Schistosome-derived omega-1 drives Th2 polarization by suppressing protein synthesis following internalization by the mannose receptor. THE JOURNAL OF EXPERIMENTAL MEDICINE 2012 Sep;209(10):1753-67. INTERNAL AND EXTERNAL COLLABORATIONS Internal: Priority area Asthma and Allergies (Asthma Mouse Models, Experimental ; Pneumology, Innate Immunity, Invertebrate Models); Priority area Infections (Biophysics, Cellular Microbiology, Clinical Infectious Diseases, Coinfection, Infection Immunology, Microbial Interface Biology, Models of Inflammation, Molecular Mycobacteriology); Medicine (Center for Clinical Studies Borstel, Clinical and Experimental Pathology) External:Ulrich Lindner, University of Lübeck; Ann-Kristin Müller, University Hospital Heidelberg; Torsten Schulze, Red Figure 3: Results of using Flow Cytometry (upper part) or (Fluorescence) Microscopy (lower part). Cross Blood Service, Mannheim; Hans-Joachim Gabius, Ludwig-Maximilians-Universität Munich IMPRESSUM FUNDING (MIO. EURO) total budget 2013 institutional funding (federal state/federal government) 17,62 j third-party funding 7,64 j of which DFG 2,18 j federal state/federal government 0,24/3,12 j EU project funding 0,39 j industry 0,99 j foundations 0,16 j miscellaneous 0,56 j total budget 2014 institutional funding (federal state/federal government) 19,9 j third-party funding 7,01 j of which DFG 1,31 j federal state/federal government 0,21/3,04 j EU project funding 1,26 j industry 1,04 j foundations 0,15 j miscellaneous 0j Leibniz Institutes are generally funded publicly in equal parts by the federal government and the federal states (Bundesländer). The FZB ist funded by the Federal Ministry of Health and the Ministry of Social Affairs, Health, Science and Equality of Land Schleswig-Holstein. ACADEMIC DEGREE AND PROFESSIONAL QUALIFICATION PATENTS AND LICENSES 2014 15 5 Diploma 1 0 Bachelor of Science 1 3 Master of Science 4 3 10 9 Dissertation Assignation of patents 7 Stock of patent families 18 Stock of licensing agreements 2013 7 Income from royalties (T j) in 2013 345 Income from royalties (T j) in 2014 25 Technicians Seite 77 / Facts & Figures • Networks CONFERENCES AND WORKSHOPS PEER REVIEWED PUBLICATIONS 2013 2014 2013 2014 29 18 124 127 GUEST SCIENTISTS BOOKS AND BOOK ARTICLES 2013 2014 national 15 10 2013 2014 international 36 33 3 11 NATIONAL NETWORKS 2013/2014 INTERNATIONAL NETWORKS 2013/2014 German Research Foundation ( DFG) 7th EU-Frame Program Exzellenzcluster ‘Entzündung an Grenzflächen’ TB-PAN-NET: Pan-European network for the study and clinical SFB/TR 22 ‘Allergische Immunantwort der Lunge’ management of drug resistant tuberculosis SPP 1313 ‘Biological Responses to Nanoscale Particles’ TheSchistoVac: The targeted development of a new generation SPP 1580 ‘Intracellular compartments as places of vaccine for Schistosomiasis pathogen-host-interactions’ PiroVac: Improvement of current and development of new GRK 1727 ‘Modulation der Autoimmunität’ vaccines for theileriosis and babesiosis of small ruminants GRK 1743 ‘Gene, Umwelt und Entzündung’ [coordination: FZB] IGRK 1911 ‘Immunregulation der Entzündung bei EDENext: Biology and control of vector-borne infections Allergien und Infektionen’ in Europe PathoNgen -Trace - Next Generation: Genome based high Federal Ministry for Education and Research resolution tracing of pathogens [coordination: FZB] Deutsches Zentrum für Lungenforschung NAREB: Nanotherapeutics for antibiotic resistant emerging Deutsches Zentrum für Infektionsforschung bacterial pathogens Schwerpunkt Biophotonik Medizinische Infektionsgenomik Others Förderinitiative NanoCare ‚Prädiktion Centre for Nanovaccines, Programkomitteen f. humantoxikologischer Wirkung synthetischer Strategiske Vaekstteknologier, Kopenhagen. Carbon Black Nanopartikel‘ ‘Imaging vaccine efficacy’ GLUTEVIS NanoCOLT: Langzeitwirkung modifizierter Carbon Black Nanopartikel auf gesunde und vorgeschädigte Lungen TB or not TB GOLD.net – Deutsches Netzwerk für diffus parenchymatöse Lungenerkrankungen ORGANIZATION CHART 03/2015 Board of Curators Chair: R. Fischer Public Relations B. Weller Scientific Advisory Board Chair: Prof. Dr. C. Vogelmeier Scientific Officer Dr. B. Brand CEO Prof. Dr. S. Ehlers Board of Directors/Extended Board CEO: Prof. Dr. S. Ehlers Program Director: Prof. Dr. U. Schaible, deputy: Prof. Dr. S. Niemann Program Director: Prof. Dr. H. Fehrenbach, deputy: Prof. Dr. F. Petersen Medical Director: Prof. Dr. P. Zabel Head of Administration: J. Repp Administration Head: J. Repp CENTRAL UNITS Central Units PRIORITY RESEARCH AREA INFECTIONS Director: Prof. Dr. U. Schaible MEDICINE Director: Prof. Dr. P. Zabel PRIORITY RESEARCH AREA ASTHMA AND ALLERGY Director: Prof. Dr. H. Fehrenbach Bioanalytical Chemistry Dr. D. Schwudke Hospital Prof. P. Zabel Clin. & Mol. Allergology Prof. U. Jappe IT M. Reinhold Human Ressources R. Niemeyer Biophysics Prof. T. Gutsmann Medical Service Center Prof. E. Vollmer Mucosal Immunology & Diagnostics PD Dr. A. Frey Finances and Purchasing n.n. Immunobiophysics PD Dr. A . Schromm Center for Clinical Studies Dr. C. Herzmann Experimental Asthma Research Prof. S. KraussEtschmann Fluorescence Cytometry Dr. J. Behrends Dr. T. Scholzen Sci. Information Services & Library R. Stief / C. Engler Techn. Building & Construction Management D. Westphal Molecular Mycobacteriology Prof. S. Niemann Clin. & Exp. Pathology Prof. E. Vollmer Experimental Pneumology Prof. H. Fehrenbach Animal Facility Dr. I. Monath Hospital Management V. Büttner Clinical Infectious Diseases Prof. C. Lange Biobank PD Dr. K. Gaede Structural Biochemistry Prof. O. Holst BBRS Dr. S. Paetzold Bioinformatics Dr. K. Fellenberg Diagnostic Mycobacteriology Prof. S. Ehlers (temp.) Biochemical Immunology Prof. F. Petersen In-Firm Training Prof. A. Petersen Microbial Interface Biology PD Dr. N. Reiling Innate Immunity Prof. H. Heine Office of Social Affairs U. Schroer Infection Immunology Dr. C. Hölscher Invertebrate Models Dr. C. Wagner Cellular Microbiology Prof. U. Schaible Mouse Models of Asthma Dr. M. Wegmann ADMINISTRATION Head: J. Repp QUALITY MANAGEMENT Equal Opportunities N. Grohmann Ombudscouncil Prof. C. Stamme PD Dr. S. MüllerLoennies Dr. T. Scholzen GSP Coordination PD Dr. A Schromm Dr. F. Schocker Biosafety Dr. J. Petersen Hazardous Subst. Dr. N. Röckendorf Radioprotection L. Lang Dr. C. Hölscher Animal Protection Dr. C. Keller Data Protection K. Clüver Health & Safety Dr. R. Birke Coinfection Dr. B. Schneider Cellular Pneumology Prof. C. Stamme Models of Inflammation Prof. G. Graßl Junior Research Groups University Liaison Groups Parkallee 1-40 23845 Borstel Tel.: +49 (0)4537.188 0 [email protected] www.fz-borstel.de
