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DAS SCHöNSTE, WAS WIR ENTDECKEN KöNNEN, IST DAS

EinbettenHerunterladen
Forschungsbericht
2011/2012
Das Schönste,
was wir entdecken
können, ist das
Geheimnisvolle
Albert Einstein
Forschungsbericht
2011/2012
Leibniz-Zentrum
für Medizin und
Biowissenschaften
Impressum
Vorwort
Seite 6
Wissenschaftlicher Beirat/Scientific Advisory Board
Prof. Dr. Klaus Dieter Pfeffer
United against Infections: German Centre for Infection Research
Seite 8
Unbeschwert atmen!
Seite 10
Borstel 2020 – fit für die Zukunft?
Seite 14
Farewell
Seite 18
Highlights
2011
Seite 22
2012
Seite 16
Direktor des Institutes für Medizinische Mikrobiologie
und Krankenhaushygiene, Universität Düsseldorf
(Vorsitzender)
Research Reports
Research Area Infections
Prof. Dr. Dr. Barbara Wollenberg
Direktorin der Klinik für Hals-, Nasen- und Ohrenheilkunde,
Universität zu Lübeck
(stv. Vorsitzende)
Prof. Dr. Axel Brakhage
Leiter des Leibniz Instituts für Naturstoff-Forschung und Infektionsbiologie e.V.
Hans-Knöll-Institut (HKI), Jena
Prof. Dr. Christoph Dehio
Infection Biology, Biozentrum, Universität Basel
Prof. Dr. Bernhard Fleischer
Vorstand des Bernhard-Nocht-Instituts für Tropenmedizin, Hamburg
Research Area Asthma and Allergy
Cellular Microbiology
Medical and Biochemical Microbiology
Immunochemistry
Biophysics
Immunobiophysics
Molecular Inflammation Medicine
Infection Immunology
Microbial Interface Biology
Models of Inflammation
Veterinary Infection Biology
and Immunology
Cellular Pneumology
28
30
32
34
36
38
40
42
44
Medical Infrastructure
46
48
Experimental Pneumology
Molecular Immunology
Biochemical Immunology
Invertebrate Models
Immune Cell-Analytics
Innate Immunity
Mouse Models in Asthma
Barrier-Integrity
Structural Biochemistry
Cellular Allergology
Clinical and Molecular Allergology
Mucosal Immunology and Diagnostics
54
56
58
60
62
64
66
68
70
72
74
76
Molecular Mycobacteriology
50
Immunobiology
78
Clinical Infectious Diseases
52
National Reference Center
for Mycobacteria
80
Clinical and Experimental Pathology 82
Center for Clinical Studies
84
Facts & Figures
Funding
Seite 86
Prof. Dr. Erika Jensen-Jarolim
Patents
Seite 86
Head of Comparative Medicine,
Academic degree and professional qualification
Seite 87
messerli Research Institute
Conferences and workshops
Seite 87
Medical University of Vienna
Guest scientists
Seite 87
Peer review publications
Seite 87
Prof. Dr. Claus Vogelmeier
Books and book articles
Seite 87
Direktor der Klinik für Innere Medizin
National networks
Seite 88
Schwerpunkt Pneumologie
International networks
Seite 88
Universitäts-Klinikum Gießen und Marburg
Administrative and technical services
Seite 89
Standort Marburg
Imprint
Seite 90
Organization Chart
Seite 91
xxxxxxxxxxxxxxxxxxxxx
Inhalt
5
MAGAZIN
Impressum
VORWORT
Schwerpunkten „Infektionen“ und „Asthma und Allergie“. Die
Der vorliegende Forschungsbericht informiert in kompakter
Konzentration auf ein einziges Organ, die Lunge, und die ganz-
Form über die wissenschaftlichen Aktivitäten des Forschungs-
heitliche Betrachtungsweise, die von molekularen Strukturen bis
zentrums Borstel in den Jahren 2011 und 2012. Der Magazinteil
hin zum komplexen biologischen System des Menschen selbst
berichtet über zukunftsweisende Umstrukturierungen sowohl im
reicht, eröffnen ein völlig neues Verständnis für Zusammenhän-
Management als auch in den Forschungsschwerpunkten des
ge. Dabei streben wir u.a. die folgenden Ziele an:
Zentrums, bewegende Ereignisse und wichtige Forschungser-
1.Identifizierung neuer Zielstrukturen für anti-bakterielle, anti-
gebnisse, Verbundaktivitäten und neue Forschungsgruppen. Ex-
entzündliche und anti-allergische Prophylaxen und Therapien.
emplarisch ausgewählte Projekte der Forschungsteams zeigen
2. Nutzung der Kenntnis von Resistenz-, Virulenz-, Persistenz- und
sowohl die Kernkompetenz auf als auch die erbrachte Leistung.
Reaktivierungsmechanismen für Therapien und Prognose.
Auf diese Weise möchten wir nicht nur versierten Kollegen und
3. Identifikation von Immunmodulatoren zur Prophylaxe und The-
Kolleginnen, sondern auch einer breiteren Öffentlichkeit unsere
rapie von infektiösen und nicht-infektiösen Entzündungen und
wissenschaftlichen Fragestellungen und Ziele näher bringen,
deren chronischer Ausprägung.
aber auch die Fortschritte aufzeigen, die wir in den beiden
Stolz sind wir auf die hervorragende Arbeit aller unserer Mitar-
vergangenen Jahren bei der Lösung gesellschaftlich relevanter
beiterinnen und Mitarbeiter, ein herzlicher Dank gilt allen Freun-
medizinischer Probleme erzielt haben.
den und Förderern aus Wissenschaft, Politik und Wirtschaft. Mit
Ihrer Unterstützung blicken wir optimistisch in die Zukunft!
Das FZB sieht seine gesellschaftspolitisch bedeutsame wissenschaftliche Mission in der grundlagen- und patientenorien-
Stefan Ehlers, Heinz Fehrenbach, Stefan Niemann, Frank Peter-
tierten Forschung auf dem Gebiet der Pneumologie mit den
sen, Ulrich Schaible, Peter Zabel
PREFACE
ma and allergies”. Putting emphasis on a single organ, the
The present report informs about the scientific activities of the
lung, and looking at it in its entirety spanning from molecular
Research Center Borstel (RCB) in the years 2011 and 2012 in
structures via cellular processes to the application of com-
a compact form. The magazine section reports on the recent
plex model systems, open up an entirely new understanding
re-organisation of both the management and the research
of causal relationships. Some of our main goals are:
priorities, events affecting the Borstel community, important
1.Identifcation of new target structures for anti-bacterial,
research results, scientific networks and new research teams.
anti-inflammatory and anti-allergic preventive and thera-
By way of examples, hand-picked projects of the research
peutic interventions.
groups and clinical units illustrate both the core competence
and the achieved accomplishments.
2.Exploiting our knowledge of mechanisms of resistance,
virulence, persistence and reactivation for improving
therapy and prognosis.
In this way, we would like to give an understanding of our
3.Identification of immune modulators for prophylaxis
research approach and objectives not only to experienced
and therapy of infectious and non-infectious, acute and
colleagues but also to a broader public. In addition, this
Focus on the Lung
S c i e n c e- , D i s e a s e- a n d
Pat i e n t- o r i e n t e d L u n g R e s e a r c h
chronic inflammatory conditions.
report shows the progress we have made during the last two
We are proud of the excellent work of all of our colleagues,
years in contributing to solving medical problems relevant
and we are deeply grateful to all friends and supporters
to society.
from science, politics, and industry. With your support we
are looking optimistically into the future!
The sociopolitical and scientific mission of the Research Center Borstel is comprehensive health and biomedical research
Stefan Ehlers, Heinz Fehrenbach, Stefan Niemann, Frank Pe-
in pneumology focusing on “infectious diseases” and “asth-
tersen, Ulrich Schaible, Peter Zabel
7
xxxxxxxxxxxxxxxxxxxxx
Vorwort • Preface
die Lunge im Fokus
Grundlagen-, krankheits- und
pat i e n t e n - o r i e n t i e r t e F o r s c h u n g
reportage
German Centre for Infection Research
by PD Dr. Stefan Niemann, Head of Molecular Mycobacteriology and Coordinator TTU Tuberkulose
Despite the availability of antibiotics and vaccines for decades,
The mission of the Thematic Translational Unit „Tuberculosis“ is
infections still represent a major cause of morbidity and mortali-
to improve TB infection control, with a focus on M/XDR-TB.
ty globally. New emerging, re-emerging and multidrug resistant
Main research areas are:
pathogens are potentially challenging and can rapidly spread
worldwide in an increasingly globalized world with free travel.
• To foster the development and application of new
In addition, the mechanisms that make particular pathogens
more virulent are only incompletely understood.
• To provide molecular surveillance of emerging MTB
... with this philosophy, the German Centre for Infection Research
(DZIF) started his work in December 2012. DZIF is one of six German
Centres for Health Research, recently initiated by the Federal Ministry for Education and Research (BMBF). The new centres will provide optimal research conditions to combat major diseases such as
infections more successfully in future.
diagnostic tests
genotypes to efficiently gauge transmission rates
The new challenges for the fight against infections require in-
• To identify and validate biomarkers that reflect
tegrative and interdisciplinary concepts that provide a platform
for a close cooperation of experts from translational research,
• To enhance the repertoire of anti-infective regimens
treatment outcome
epidemiology and clinical practice. To address this need, DZIF
combines expertises and infrastructures of 32 partners located
Translational research of the TTU-TB encompasses the deve-
at six sites throughout Germany to better coordinate and stra-
lopment, refinement and clinical implementation of new diag-
tegically align translational infection research in Germany and
nostics, new drugs, improved patient management as well as
develop new measures/tests for improved diagnostics, preven-
improved TB control via molecular epidemiology.
tion and treatment of infectious diseases.
For local and global control of TB, precise epidemiological
PD Dr. Stefan Niemann
To achieve this, DZIF has formed Thematic Translational Units
data are necessary that must include in-depth molecular inves-
(TTUs) of scientists, each dedicated to one specific pathogen
tigations to unravel transmission dynamics and hotspots. Lon-
or infectious disease: Emerging Infections, Tuberculosis, Mala-
gitudinal implementation of web-based molecular surveillance
ria, HIV, Hepatitis, Gastrointestinal Infections, Infections of the
is required to evaluate the success of implemented changes in
Immunocompromised Host, Healthcare-associated, Antibiotic-
diagnostic and control strategies. However, translation is also a
resistant Bacterial Infections and Novel Antiinfectives. The work
reciprocal process: basic laboratory research must be focused
of the TTUs is supported by six infrastructural platforms (e.g.
on questions that arise in the clinic and adapt its methods to
Product Development, Clinical Trial Unit, and African Partner
optimize and validate biomarkers and treatment regimens.
Sites) and accompanied by an innovative and highly attractive
educational program for students and postgraduates that aims
Increasing rates of M/XDR Mtb strains (more than 30 % of never-
at attracting young medical doctors and scientists into infection
before-treated patients in some areas) threaten to substantially
research. By exchange programs, structured doctoral programs
reduce the cure rate of TB. According to recent meta-analyses,
and collaborative spring and autumn schools, the DZIF Acade-
treatment of XDR-TB and MDR-TB is on average only successful
my reflects and emphasizes the synergistic nature of DZIF, and
in 40-60 % of patients. M/XDR-TB spread is accelerated by the
shares its mission to close the gap between basic research and
development of particular „super transmissible“ M/XDR strains
clinical development.
suggesting increased virulence and/or infectivity of these. The
TTU-TB effort will focus on these relatively recent threats to TB
In the framework of the DZIF, FZB is part of the partner site
control. To this end, molecular tools including next generation
Hamburg-Lübeck-Borstel and is coordinating the TTU Tubercu-
sequencing will be further developed and linked to clinical
losis (TTU-TB) with four participating DZIF focus sites (Hamburg-
risk assessment and the public health sector in Germany (and
Lübeck-Borstel, München, Hannover-Braunschweig,Tübingen)
NGOs elsewhere) to improve diagnosis, treatment and infection
and two external partner sites (Münster, Freiburg).
control.
With 8 million newly infected individuals and 1.4 million deaths
In a longitudinal perspective, TTU-TB therefore aims to strate-
every year, TB is still a major global health threat. The problem
gically transform the existing expertise in German TB research
is compounded by HIV/TB co-infections and the emergence of
into a DZIF National Center for Tuberculosis with translational
multi- and extensively drug-resistant (MDR and XDR) strains of
focus and international outreach.
Mycobacterium tuberculosis complex, particularly in Eastern Europe, Sub-Saharan-Africa and Asia. TB control, on the global
level, is faced with several challenges: There is currently no vaccine that efficiently protects against pulmonary TB in adults; the
arsenal of anti-TB drugs is limited and there are only few new
drugs in industrial pipelines; biomarkers to predict or monitor
treatment success are virtually absent; and access to TB diagnostics is restricted in resource-poor settings.
9
German Centre for Infection Research
United against Infections:
reportage
Mit der Einrichtung der neuen Nachwuchsgruppe Mausmodelle
(ALI), Diffuse parenchymatöse Lungenerkrankungen (DPLD), Pul-
des Asthma (Leiter: M. Wegmann) am Forschungszentrum Borstel
monale Hypertonie (PH), Lungenerkrankungen im Endstadium
zum Oktober 2012 und in Zusammenarbeit mit der Forschungs-
(ELD) und Lungenkrebs (LC). Die krankheitsorientierte Forschung
gruppe Experimentelle Pneumologie wird das Ziel verfolgt, im
wird durch Plattformen für Biobanking, Lungenbildgebung (Ima-
Rahmen und in Kooperation mit Vertretern anderer Standorte des
ging), Klinische Studien und Forschungsmanagement unterstützt.
DZL (Hannover, Heidelberg, Gießen / Marburg) neuartige, auch
Basierend auf der Tatsache, dass es substantielle mechanistische
transgene, phänotyp-spezifische Mausmodelle für mechanistische
Überschneidungen bei Initiation und Fortschreiten dieser Lun-
wie präklinische Therapie-Studien zu entwickeln. Den derzeit na-
generkrankungen gibt, verfolgt das DZL einen interdisziplinären
hezu ausschließlich einen TH2- / Eosinophilen-dominierten Asthma-
und integrierten wissenschaftlichen Ansatz, der die dynamischen
Phänotyp reflektierenden Modellen stehen keine entsprechenden
Beziehungen zwischen Entwicklung, Regeneration und Reparatur
Modelle zu tierexperimentellen Studien an anderen Phänotypen,
im Rahmen von Lungenerkrankungen, sowie Mechanismen der
wie zum Beispiel des Neutrophilen- oder TH 17-dominierten bzw.
Entzündung (infektiös und nicht-infektiös) und benigner und malig-
Kortikosteroid-resistenten Asthma, zur Seite.
ner Hyperproliferation einbindet.
Im dem weiten Feld der Erforschung der zellulären Mechanismen,
Das Forschungszentrum Borstel hat die Schwerpunkte seiner Ak-
die für Pathogenese, Diagnostik und Therapie distinkter Asthma-
tivitäten den Krankheitsbereichen Asthma & Allergie, COPD und
Phänotypen von Bedeutung sind, widmen sich die im DZL aktiven
Lungenkrebs gewidmet und ist außerdem prominent in den Platt-
Forschungsgruppen Strukturbiochemie, Klinische und Molekulare
formen Lungenbildgebung und Biobanking vertreten. Das For-
Allergologie, Angeborene Immunität, Biochemische Immunolo-
schungszentrum Borstel ist dabei an der Umsetzung folgender
gie, Experimentelle Pneumologie sowie Mukosale Immunologie
für die jeweiligen Krankheitsbereiche und Plattformen DZL-weit
und Diagnostik folgenden Schwerpunktaspekten: der Identifizie-
definierten Meilensteine beteiligt:
rung struktureller und funktioneller Eigenschaften von Allergenen,
die zu qualitativ unterschiedlichen Immunreaktionen führen kön-
Prof. Dr. Heinz Fehrenbach
Von Prof. Dr. Heinz Fehrenbach, Programmdirektor Asthma und Allergie
Das Deutsche Zentrum für Lungenforschung (DZL) hat sich der
Mission der Stärkung der translationalen Forschung zur Aufklärung von Krankheitsentstehung und Verbesserung der Diagnostik
und Therapie häufiger Lungenerkrankungen verschrieben.
Krankheitsbereich Asthma & Allergie
nen (dimer/oligomer formation; epitope mapping); der Charak-
Am Aufbau eines Patientenregisters zu Allergien und Asthma
terisierung der Rolle des Atemwegsepithels für die Entstehung
und der umfassenden klinischen Charakterisierung der einge-
distinkter Asthma-Phänotypen (epitheliale Signaturen); der Iden-
schlossenen Patientengruppen, insbesondere mit dem Ziel ado-
tifizierung einzelner Gene und Signalwege in Geweben der Epi-
leszente und erwachsene schwere Asthmatiker von Patienten
thelial-Mesenchymal-Trophic-Unit (EMTU) mit Schlüsselfunktionen
mit anderen Erkrankungsphänotypen abgrenzen zu können, sind
bei der Asthma-Pathogenese (remodelling, bronchoconstriction);
die Medizinische Klinik, das Studienzentrum und die Interdiszip-
der Analyse der Bedeutung des angeborenen Immunsystems bei
linäre Allergie-Ambulanz beteiligt. Daneben sind an der Samm-
der Entstehung distinkter Asthma-Phänotypen; der Identifizierung
lung und Lagerung von Biomaterialien für die Hochdurchsatz-
phänotyp-spezifischer Komponenten des adaptiven Immunsys-
verfahren zur detaillierten Charakterisierung der verschiedenen
tems (imprinted phenotypes, cell differentiation, role of specific
Asthma-Phänotypen sowie der Etablierung von Isolations-und
cell subtypes) sowie der Identifikation neuer Biomarker und mole-
Kulturtechniken zu experimentellen Studien nasaler und bron-
kularer Targets für Asthma-Phänotypen.
chialer Epithelzellen in Kooperation mit dem Standort München
die Forschungsgruppe Klinische und Experimentelle Pathologie
Krankheitsbereich COPD
einschließlich der mit Hilfe des DZL aufgebauten Biobank enga-
Im Arbeitspaket COPD-1 (Remodellierung, Regeneration und Re-
giert. Ihr obliegt auch die Harmonisierung der etablierten Tech-
paratur: Von Tiermodellen bis zu menschlichen Gewebeproben)
niken, die gleichermaßen für die Krankheitsbereiche COPD und
widmet sich die Forschungsgruppe Klinische und Experimentel-
Innerhalb des seit Oktober 2011 vom Bundesministerium für
das Forschungszentrum Borstel, die Universitäten Lübeck und
Lungenkrebs von Bedeutung sind. Die molekulare Analyse soll
le Pathologie der Validierung von Kandidatengenen in nativen
Bildung und Forschung im Rahmen der Deutsche Zentren der
Kiel, das Universitätsklinikum Schleswig-Holstein sowie die Lun-
mit dem Vergleich epithelialer Transkriptome im Grenzbereich
Geweben und Primärzellkulturen von COPD-Patienten sowie der
Gesundheitsforschung geförderten DZL-Konsortium arbeiten
genclinic Großhansdorf zusammengeschlossen haben.
schweres Asthma versus COPD helfen, Unterschiede bei deren
Transkriptomanalyse und Targetvalidierung an humanen Proben
Pathogenese besser zu erkennen und damit gegebenenfalls zu
(Sputum, Lungengewebe). Hier besteht eine enge Verzahnung
zahlreiche universitäre und außeruniversitäre Institutionen an
folgenden Standorten zusammen: München (CPC-M), Heidel-
Die Forschungsaktivitäten innerhalb des DZL konzentrieren sich
einer verbesserten Diagnostik und damit einer klareren Abgren-
mit den beim ARCN-Partner Lungenclinic Großhansdorf angesie-
berg (TLRC-H), Gießen/Marburg (UGMLC), Hannover (BREATH)
auf folgende acht Krankheitsbereiche: Asthma und Allergie (AA),
zung und zugeschnittenen Therapie der beiden Krankheitsbilder
delten Projekten und eine ausgeprägte, auch für den Krankheits-
und das Airway Research Center North (ARCN), zu dem sich
Chronisch obstruktive Lungenerkrankungen / Emphysem (COPD),
bzw. -phänotypen zu gelangen.
bereich Asthma & Allergie überaus dienliche Vernetzung bei der
11
Deutsches Zentrum für Lungenforschung
Unbeschwert Atmen!
Zystische Fibrose (CF), Pneumonie und akutes Lungenversagen
Sammlung von Biomaterialien. Ebenfalls ein hohes Potential an
gegeben und, im Rahmen von Laboraufenthalten, praktische
hochwertiger und gesicherter Qualität sowie die datenschutzkon-
tung in die DZL-Strukturen federführend vom Forschungszentrum
Krankheitsbereiche übergreifender Bedeutung besitzt das von
Anleitungen zum Einsatz von state-of-the-art Techniken angebo-
forme Speicherung der damit verbundenen Daten und Informati-
Borstel wahrgenommen wird, soll die Verfügbarkeit von Bioma-
der Forschungsgruppe Mukosale Immunologie und Diagnostik
ten werde. Die Forschungsgruppe Experimentelle Pneumologie
onen. Mit Hilfe der BMB-NORD, deren Organisation und Einbet-
terialproben und assoziierten Daten durch gemeinsame Nutzung
im Arbeitspaket COPD-2 (Biomarker und Phänotypen) verfolgte
führt solche im Rahmen des DZL angebotene, problemorientier-
Projekt zur Entwicklung von Mucin-reaktiven Sonden.
te Kurzlehrgänge durch.
Krankheitsbereich Lungenkrebs
Plattform Biobanking
Die Forschungsgruppe Klinische und Experimentelle Pathologie ist
Das DZL hat die Schaffung von Biobank-Infrastrukturen als un-
in das Arbeitspaket LC-2 (Maligne Transition vom Bronchialepithel
abdingbare Grundlage der translationalen Forschung erkannt.
zum Karzinom) eingebunden, das zum Ziel hat, epitheliale Signatu-
Deshalb werden sowohl die Einrichtung bzw. der Ausbau lokaler
ren zu identifizieren und damit einen die Krankheitsbereiche Asth-
Infrastrukturen als auch die Schaffung eines zentralen DZL-Bio-
ma & Allergie, COPD und Lungenkrebs überspannenden Bogen
bankenportals aller Standorte gefördert. Mittlerweile ist das DZL
schlägt. Auch in diesem Krankheitsbereich konnte eine aufs engste
und damit jeder Principal Investigator des DZL Mitglied der Tech-
ausgeprägte inhaltliche und strukturelle Vernetzung mit dem ARCN-
nologie- und Methodenplattform für die vernetzte medizinische
Partner Lungenclinic Großhansdorf aufgebaut werden.
Forschung e. V. (TMF) und kann somit deren Angebote nutzen.
optimiert sowie die Voraussetzungen für die translationale Forschung verbessert werden. Damit wurden in kurzer Zeit Infra-
AA DPLD
ELD
COPD
CF
PH
ALI LC
FZB-Mitgliedern des Deutschen Zentrums für Infektionsforschung
strukturen geschaffen, die nicht nur den translational orientierten
Projekten im Rahmen des DZL zugutekommen sondern auch den
zur Nutzung offen stehen. Organisatorisch ist die BMB-Nord der
Hyperproliferation
Forschungsgruppe Klinische und Experimentelle Pathologie zuge-
benign
malignant
ordnet. Zur Sicherung optimaler Bedingungen für die Gewinnung
non-infectious
infectious
der wertvollen Biomaterialien ist die BMB-Nord hinsichtlich ihrer
Disease cohorts, epidemiology, helath care
Aktivitäten bei der Materialsammlung und Datenerhebung in die
“Deep phenotyping“, clinical pilot trials, phase II/III trials,
exploitation and implemention
zeitig die unabdingbare räumliche Nähe zu Routinelabors und
Biobanking, human repositories, biomarker studies
Studienzentrum gewährleistet. Mit diesen im Rahmen der DZLAktivitäten geschaffenen Strukturen ist es gelungen, am FZB eine
13
Forschungslabore der Medizinischen Klinik integriert, was gleich-
Plattform Lungenbildgebung
Um den besonderen ethischen und datenschutzrechtlichen An-
Preclinical trials and pulmonary pharmatherapy
Vorherrschendes Ziel der Plattform ist es, den Einsatz von Ima-
forderungen an eine solche Biobank Rechnung zu tragen, haben
Pathophysiology and disease models
Ausgangslage zu schaffen, die das Forschungszentrum Borstel
ging-Technologien in den verschiedenen Disease Areas auf ra-
die ARCN-Partnerinstitutionen Ende 2012 einen Kooperationsver-
Developmental and system biology
in idealerweise in die Lage versetzt, translationale Forschungs-
diologischer und mikroskopischer Ebene zu steuern. Der Bereich
trag geschlossen, der die geplante Zusammenarbeit im Rahmen
Functional genomic and cellular phenotypes (incl. stem cells)
ansätze weiter auszubauen und sich damit gut gewappnet zu
Mikroskopie, der vom Forschungszentrum Borstel gemeinsam
der Gründung und des Betriebs der sogenannten Biomaterial-
Molecular signatures and target structures
wissen, den künftig steigenden Anforderungen der translationale
mit dem Standort Hannover koordiniert wird, ist vorwiegend ex-
bank Nord (BMB-Nord) regelt. Zielsetzung ist die Schaffung einer
Konzertierte, erkrankungsübergreifende Strategie der translationalen
perimentell ausgerichtet. Hier sollen zum einen Empfehlungen
Sammlung und Lagerung von Biomaterialproben in möglichst
Forschung des Deutschen Zentrum für Lungenforschung
Gesundheitsforschung gerecht zu werden.
Deutsches Zentrum für Lungenforschung
Inflammation
Resolution
Generation
Regeneration
Repair
Interview
Borstel 2020 – fit für die Zukunft?
„gut“ ist, wenn die Thematik nicht in der gebotenen Breite und Tiefe bearbeitet werden
kann.
Das Forschungszentrum Borstel wurde 1947 als Tuberkulose-Institut mit der Unterstützung des Landes SchlesFührt diese Spezialisierung nicht zu einem
Zäsur, die zur Neustrukturierung des Institutes in der Etablierung von zwei wissenschaftlich grundlagenori-
Verlust an Flexibilität?
entierten Bereichen und einem wissenschaftlich klinisch ausgerichteten Bereich führte. Jeder Bereich war mit
Wenn Flexibilität bedeutet, dass man jeden
einer C4-Professur an den Universitäten Kiel und Lübeck verbunden. 1981 wurde die erste Forschungskonzepti-
Tag neu entscheiden kann, woran man wis-
on mit der Gesamtthematik Infektion und Immunbiologie erarbeitet, die in den folgenden Jahren stetig weiter
senschaftlich arbeiten möchte - ob an der
entwickelt wurde und 1997 den Schwerpunkt Allergie und 2000 die Thematik Tumorbiologie in die Konzeption
Tuberkulose oder an der Darmflora der
aufnahm.
Ameise - dann stimmt das natürlich. LeibnizEinrichtungen verstehen sich jedoch als Stra-
Der zweite markante Einschnitt in Struktur und wissenschaftlicher Ausrichtung des Zentrums erfolgte in den
tegie-Forschungszentren, deren Mission je-
letzten 2 Jahren: Reduktion auf zwei wissenschaftliche Schwerpunktbereiche (Infektionen; Asthma & Allergie)
weils die Bearbeitung einer gesellschaftlich
und eine völlig neue Leitungsstruktur.
relevanten Fragestellung ist. Innerhalb dieses Forschungskonzepts ist Borstel völlig frei
und flexibel - und da die Ressourcen nicht
Prof. Ehlers, benötigte der Oldtimer
Tuberkulose- und Asthma-Forschung, nur
lichen Lungenerkrankungen - die die
durch missions-fremde Aktivitäten gebunden
Borstel eine Verjüngungskur oder was
konsequent – was wirklich neu war, war
Auswahl des methodischen Zugangs,
sind, sogar flexibler und Potential-reicher als
war der Auslöser für diese nicht uner-
die Abschaffung herkömmlicher Abteilun-
aber auch den Grad der notwendigen
ohne Fokussierung. Borstel kann sich jeder-
heblichen Veränderungen?
gen und die Ausrichtung der Forschung
Interdisziplinarität bestimmt. Borstel hat
zeit durch programmatische Zusammenar-
Mit 65 muss man sich entscheiden, ob
in translational angelegten Handlungs-
sozusagen, nach den frühen Jahren der
beit verstärken – hierzu haben wir das neue
man schon zum „alten Eisen“ gehört und
strängen, d.h. die weitgehende Integra-
eher phänomenologischen Untersuchung
Instrument des VIP-Fonds geschaffen, einer
den Dingen seinen Lauf lässt oder ob
tion von Forschungsgruppen mit Grundla-
von Erregerspezies und Antibiotika-Wirk-
Vernetzungs- und Innovationsförderung, die
man als „Best Ager“ noch einmal eine
gen-, präklinischer und klinischer Agenda
samkeiten, in den 80er und 90er Jahren
in jedem Programmbereich die besten Ini-
Standortbestimmung und Neu-Definition
in Programmbereiche. Hier ist Borstel
die molekular- und strukturbiologische
tiativen unterstützt, um die Gesamt-Mission
seiner
vor-
wirklich Wegweisendes gelungen, so-
„Schleife“ genommen, um sich molekül-
nach vorne zu bringen.
nimmt. Die Evaluation der gesamten In-
zusagen „Theoria cum praxi“ bis in die
orientiert der Aufklärung und Modulation
stitution durch die Leibniz-Gemeinschaft
Organisationsstruktur hinein!
von Entzündungsprozessen zu widmen.
Grundlegende Änderungen in der Lei-
Dabei hatte sich das Zentrum ein wenig
tungsstruktur sind ja kein Kinderspiel –
Entwicklungsmöglichkeiten
im Mai 2012 war der Anlass, auch im
Hinblick auf die vorhandenen begrenzten
Könnte man sagen Borstel hat den Re-
von der Kernthematik „Lunge“ entfernt
das erfordert Mut und Respekt aller Be-
Fördermittel und bei dem gegebenen
set Knopf gedrückt und kehrt zurück zu
und sehr diversifiziert. Zwischen 1995
teiligten. Selbst wenn alle guten Willens
umfassenden Sanierungsbedarf der ge-
seinen wissenschaftlichen Wurzeln?
und 2005 wurde dann das Forschungs-
sind, gibt es doch Faktoren wie Macht
samten Liegenschaften des Forschungs-
In gewisser Weise schon, da ja der An-
profil wieder geschärft, die Tuberkulose
und hierachische Ansprüche zu überwin-
zentrums eine neuerliche Fokussierung
lass zur Gründung des Forschungsinstituts
wieder stärker in den Mittelpunkt gestellt
den. Wie ist das Ihnen und Ihren Kollegen
des Forschungsprofils auf das Machbare
im Jahre 1947 die drängenden Probleme
und die Entzündungsthematik auf Asthma
gelungen?
zu versuchen. Die Auswahl der Schwer-
bei der Bekämpfung der Tuberkulose
und Allergien begrenzt. Die Trennung von
Das war ein längerer, natürlich nicht konflikt-
punkte war, angesichts der herausragen-
waren. Auch heute ist es wieder die kli-
der Tumorbiologie als Schwerpunkt folg-
freier Prozess. Dabei haben wir gelernt, dass
den Erfolge bei der langfristigen Dritt-
nisch relevante Fragestellung - allerdings
te der Erkenntnis, dass „weniger mehr
der Verlust von Macht dann kompensiert
mittelförderung vor allem im Bereich der
für ein breiteres Repertoire an entzünd-
ist“ – dann nämlich, wenn „mehr“ nicht
werden kann, wenn „kollektive Weisheit“
15
Borstel 2020 – fit für die Zukunft?
wig-Holstein und des Kreises Segeberg gegründet. In den letzten 65 Jahren gab es zwischen 1978 und 1980 eine
den, das sowohl die Überprüfung, ob die
nen Anlagen, seinen Möglichkeiten zu
schon ein paar Strategien entwickelt,
tion und effektive Organisation schaffen
gesamte Wohnanlage auf dem Gelände
machen. Borstel lebt von der inhärenten
die Bodenhaftung nicht zu verlieren (der
kann und damit insgesamt größere Hand-
erhalten werden kann und muss, bein-
Motivation seiner Mitarbeiterinnen und
Tag der offenen Tür des ZD gehört dazu,
lungsspielräume eröffnet. So haben wir die
haltet als auch die Analyse des Sanie-
Mitarbeiter, seiner umfassenden Solida-
auch der regelmäßige Besuch in allen
Quadratur des Kreises geschafft: soviel Zen-
rungsbedarfs des Forschungsgebäudes
rität im Bewusstsein des gemeinsamen
Forschungsgruppen). Ich habe praktisch
tralismus wie nötig bei maximaler Kollegiali-
Parkallee 22. Natürlich dürfen wir dabei
Ziels, seiner Kooperationswilligkeit und
alle aktiven Forschungstätigkeiten auf-
tät; keiner kann alleine bestimmen, aber es
nicht vergessen, dass auch zukunftswei-
seiner Innovationsfreude. Borstel spielt
gegeben und erlerne jetzt das Manage-
muss auch kein Vollkonsens in allen Belan-
sende Investitionen, z. B. die Errichtung ei-
zwar in der Forschungs-Bundesliga, ist
ment-Handwerk, u.a. werde ich hierzu
gen erreicht werden. Selbstverständlich sind
ner sicherheitstechnisch höchstmodernen
aber zufrieden, nicht zu den ersten 3
auch diverse Schulungen besuchen. Hier-
auch heute die Kollegiumssitzungen nicht
Anlage für Forschungen mit multi- und
bis 5 zu gehören - weil man sich dann
bei stelle ich fest, dass die Leitung eines
frei von Rivalitäten und Machtansprüchen.
extremresistenten Tuberkulose-Erregern,
zu stark verstellen und „dem Markt“ an-
Zentrums von der Größe Borstels wirklich
Die verschiedenen Interessenlagen müssen
die der strategischen Standortsicherung
gleichen müsste. Borstel ist manchmal
nicht „nebenher“ gemacht werden kann
sich heute allerdings zum Gesamtwohl des
dienen, notwendig werden. Insgesamt
ein bisschen wie das gallische Dorf bei
– es gibt viel zu viele Baustellen, die alle
Zentrums in Programmbereich-übergreifen-
muss auch bedacht werden, dass trans-
Asterix und Obelix – sehr originell, ein
die ungeteilte Aufmerksamkeit und einen
den Abstimmungen durchsetzen, und hier
lationale Forschung unter Einbeziehung
wenig verschroben, ein wenig kauzig,
grundlegenden Lösungsansatz benöti-
sind gute Argumente und überzeugende,
von Patienten der Medizinischen Klinik
aber liebenswert und siegesgewiss. Und
gen. Es wird sicher Jahre brauchen, hier
Konzepte sowie schriftliche Beschlussvor-
erhebliche Mehrkosten verursacht, die im
den Zaubertrank brauen wir Borstelerin-
in allen Bereichen wieder Ruhe hinein
lagen erforderlich. Vor allem aber ist ent-
Budget realistisch veranschlagt werden
nen und Borsteler eben einfach immer
zu bringen und nicht im Tagesgeschäft
scheidend, wie groß das Vertrauen der Kol-
müssen. Bei diesen Planungen müssen
gemeinsam!
zu versinken. Auf jeden Fall habe ich
legiumsmitglieder untereinander ist – und
wir dem Wissenschaftsministerium des
es wächst immer dann, wenn jemand auch
Landes Schleswig-Holstein in Verhandlun-
Prof. Ehlers, ein persönliches Wort
gegenteiliger Behauptungen – viel über-
mal auf seinen eigenen unmittelbaren Vor-
gen treten, um Sondertatbestände auszu-
zum Schluss: Sie haben für die Posi-
sichtlicher und planbarer ist als das Ma-
teil zu verzichten bereit ist.
weisen, die eine zusätzliche Finanzierung
tion des Zentrumsdirektors Ihre For-
nagen eines Zentrums!
einsehen müssen, dass Forschen – trotz
erforderlich machen. Soll heißen: Sparen
schung komplett aufgegeben. Sie
Um Borstel fit für die Zukunft zu machen,
ja, zum Teil sogar schmerzhaft, aber nicht
waren Impulsgeber in wichtigen Ver-
Ich bin natürlich dabei, einen neuen Kreis
müssen Sie das Zentrum sozusagen als
um jeden Preis, bei gleichzeitigem selbst-
bundstrukturen wie dem Exzellenz-
von Vertrauten aufzubauen, zu denen
Quasi-Sanierer mit einer heilsamen Fas-
bewussten Einfordern von Unterstützung
cluster oder dem Deutschen Zentrum
auch die mir unmittelbar zugeordneten
tenkur verordnen?
durch die Geldgeber dort, wo sie – zu
für Infektionsforschung, haben erfolg-
Referate und Stabsstellen gehören. Aus
Fasten muss nicht an jeder Stelle sein; es
Recht! – stolz auf die Exzellenz des For-
reich das BMBF Konsortium TB or not
diesem Kreis habe ich auch richtungs-
handelt sich eher um ein Fitness-Programm!
schungszentrums verweisen können.
TB geleitet – wird Ihnen all das nicht
weisende Geschenke erhalten, z.B. von
fehlen oder ist die Verwandlung vom
meiner persönlichen Referentin das Buch
Borstel hat in der Vergangenheit einen
enormen wissenschaftlichen Aufschwung
Was ist Ihrer Meinung nach das Le-
Wissenschaftler zum Wissenschafts-
„Die Königsstrategie: Management für
genommen – und dabei die nachhaltige Er-
benselixier von Borstel?
manager bereits gelungen?
Männer“, das mir wohl die Augen in Sa-
neuerung der Infrastruktur, was die Bausub-
Die Menschen, die hier arbeiten! Bors-
Noch fühle ich mich nicht so fest im Sat-
chen Selbstbehauptung öffnen soll. Als
stanz aber auch die zeitgemäße Umsetzung
tel hat aus seinem scheinbaren Stand-
tel wie ich das zu meiner Zeit als For-
ich jetzt ein weiteres Management-Buch
daten-, arbeits-, und sicherheitsschutzrecht-
ortnachteil einen sichtbaren Pluspunkt
schungsgruppen- oder Bereichsleiter war.
erstanden habe mit dem bezeichnenden
licher Vorschriften betrifft, sträflich vernach-
gemacht: Man arbeitet gerne dort, wo
Auch habe ich natürlich meine unmittel-
Titel „Führen aus der Hängematte“, habe
lässigt. Hier muss ein Gesamtkonzept (ich
man selbst das Schicksal in der Hand
bare „Support Group“ verloren - es ist et-
ich jedoch erstmals wirklich begriffen, wo-
nenne es mal: „Borstel 2020“) erstellt wer-
hat, das Beste aus seiner Situation, sei-
was einsam an der Spitze! Aber ich habe
hin die Reise noch gehen muss ...
Ein Interview mit dem Zentrumsdirektor
Prof. Dr. Stefan Ehlers
geführt von Dr. Bettina Brand
17
Borstel 2020 – fit für die Zukunft?
eine transparente Entscheidungslegitima-
FArewell
Helmut Brade
Ernst Brandt
by Ulrich Schaible and Sven Müller-Loennies
by Frank Petersen
In 2012, Helmut Brade retired from his long term engagement at the Research Center Borstel. Helmut joined
On February 2011, Prof. Dr. rer. nat. Ernst Brandt retired. Prof. Brandt started his work at the Research Center
the Center in 1986 and was accredited as medical specialist in Microbiology and Infection Epidemiology.
Borstel in 1982 and his research group, the division of Biological Chemistry, was one of the „founding mem-
His prime interest was to understand the biology of endotoxins of Gram-negative bacteria, their biosynthe-
bers“ of the Department of Immunology and Cell Biology established at the Center two years before. Under
sis and interactions with antibodies. Together with the excellent research team “Medical and Biomedical
the initial direction of Prof. Hans Dieter Flad, he was able to establish a highly successful group in which
Microbiology” and many international collaborators he dug deep into the structural requirements for
numerous papers have been published and ten PhD-students have reached their degree under his supervision.
the recognition of bacterial carbohydrates by antibodies contributing to more than 240 internationally
After initial work on Interleukin-2, his discovery of a platelet-deri-
Due to his achievement, today platelets are no longer seen as
Helmut‘s group contributed significantly to the identification
me Professor of Immunochemistry and Medical Microbiology
ved isoform of β-thromboglobulin acting as a neutrophil activator
simple mediators of blood coagulation but as important players
of the first and so far only crossreactive endotoxin-neutralizing
in 1997. To bring science to young people he was a founding
(neutrophil-activating peptide 2; NAP-2; later termed as CXCL7)
in the initiation and maintenance of the innate immune response.
antibody, WN1222-5, mimicking the Toll-like-receptor 4 binding
member of the laboratory for school children “FuSch”.
was a major breakthrough on the field of chemokine research.
Ernst Brandt continued his work on the biology of thrombocyte-de-
On occasion of his retirement, a farewell-symposium was hold
researcher Helmut was always driven by his vision to improve
Helmut always assumed responsibilities for the Center. To
rived chemokines becoming a specialist here with international
at the Research Center Borstel on February 18, 2011. Many
human health. Consequently, he translated his basic research to
strengthen ethical good scientific practice, he acted as om-
reputation. Beside chemokine functions, he investigated the regu-
friends, colleagues and former PhD-students of Ernst Brandt
develop novel diagnostic tools. Thus, he identified the structural
budsman for “Self control in science” between 2005 and 2009.
lation of the corresponding receptors as well as their intracellular
could say good-bye to an outstanding scientist whose work was
epitope for the bacterial pathogens of the Chlamydia familiy to
Finally, Helmut acted as temporary director of the Department
signaling. During the last ten years, the regulatory crosstalk of
formative for the department. However, they welcomed a highly
develop an ELISA for the detection of these important infections.
of Infection Biology between 2005 and 2008.
mast cell proteases with the platelet chemokine network in aller-
motivated emeritus who will stay always in close contact with
gic diseases became a new focus in his work.
science and the Research Center Borstel.
site for lipopolysaccharide. As a medical doctor and dedicated
Always aware of the commercial potential of his research,
Helmut follows his wife and longtime collaborator, Lore, to enjoy
Helmut founded the first start-up out of the Research Center, the
their retirement together. We fullheartedly thank Helmut for his
“GlycoBioTech GmbH”. Helmut‘s excellent research was also
long-term committment for the Research Center. May Lore and
instrumental in the acquisition of a large body of grant money
Helmut enjoy a long lasting, rich and happy retirement.
and networks including the highly successful SFB 470.
Helmut was also a very active teacher and mentor. He supervised many students and lead them to successful doctoral theses and lectured at the University of Lübeck, where he beca-
Jan-Eric Ehlert, Luc
Härter, Ernst Brandt
, Andreas Ludwig,
Frank Petersen (v. l
i. n. re.)
19
Farewell
acclaimed publications.
FArewell
Buko Lindner
Ulrich Zähringer
by Ulrich Seydel und Thomas Gutsmann
by Otto Holst
After having obtained his diploma in physics at the University of Hamburg, Buko Lindner joined the Divison of
about 32 years. It was in November 1980 when Uli started his work in Borstel (then still “Forschungsinstitut
Applied Radioactivity (head Prof. J. MeiSSner) at the Research Center Borstel in 1975 and in 1976 he started to work
Borstel”), as Head of the Division of Immunochemistry. Before, he had studied chemistry at the University of
on his dissertation, which he finished in 1981. Meanwhile, the divison was re-named as Div. of Biophysics, and Prof.
Freiburg i. Br. and had had a short postdoc time at the Freiburg Max-Planck-Institute of Immunobiology. .
In September 2012, Prof. Dr. rer. nat. Ulrich Zähringer retired, having worked at Research Center Borstel for
U. Seydel was the successor of Prof. MeiSSner (1979). At that time, leprosy was in the center of interest in Borstel.
Sponsored by the German Leprosy Relief Association, a new technique was installed in the lab, the laser microproBeing a chemist, Uli started to work on the structure-function
It was not “only” lipid A which fascinated Uli, but also the other
teria. Buko Lindner and U. Seydel started a project to determine the influence of therapeutics on Mycobacterium
relation of lipid A, the endotoxic active moiety of Gram-negative
LPS moieties, i.e core and O-specific polysaccharide, and other
leprae during therapy by isolating the bacteria from skin biopsies of patients at certain intervals of treatment by
bacterial endotoxin (lipopolysaccharide, LPS), a field to which
cell envelope lipids, on which he again worked with great suc-
determining the intracellular Na+/K+-ratio of about 100 single bacteria at each time point. From the shift of the
he has significantly contributed over all these years. In the early
cess. Later he became additionally interested in Gram-positive
median the success of the treatment could be determined.
days, he together with the groups of Helmut Brade and Ulrich
bacterial cell envelope molecules, and in particular his work on
Schade had his laboratory in Parkallee 3a, where everybody
lipoteichoic acids is just outstanding.
suffered from little space. However, planning the new laboraNow, LAMMA was not only a microprobe, but also a mass spec-
versity of Kiel.Because mass spectrometry had become an increa-
tory building in Parkallee 22 was underway, and Uli was hea-
Until the mid-1980ies, structural analysis of carbohydrates used
trometer, however, with laser ionization. Consequently the two
singly important methodology for Borstel and new instrumentation
vily involved in it, thinking of and introducing a lot of details
to be performed by wet chemistry. However, mass spectrome-
persons tried to use it for structure analysis of organic molecules.
with larger mass ranges and further specifications had meanwhile
which were a prerequisite for a successful scientific work. After
try and NMR spectroscopy had been introduced as analytical
Meanwhile endotoxin came into the focus of larger parts of the
been developed, two new instruments were established: a MALDI-
opening this building in 1986, everybody was simply delighted
means around 1980, and both turned out to be extremely useful
Research Center. Consequently, they tried to get information on
TOF and later a FT-ICR mass spectrometer, which led to a large
about its construction and the facilities, many of which were
tools. It was also Uli who made sure that RCB bought a new
the chemical structure of lipid A. With success! At that time it was
number of collaborations. Now Buko is retiring, and that is the time
based on Uli’s planning.
360 MHz NMR spectrometer in 1988 and who introduced and
the largest molecule ever been mass spectrometrically analyzed.
for a short appraisal: Buko was always appreciated as a very in-
From then on Buko Lindner focused completely on organic mass
telligent, successful, and internationally respected scientist and an
From the very beginning in Borstel, Uli has been having a very
spectroscopy, later powered by a second sabbatical at the Uni-
spectrometry. He analyzed uncountable natural as well as synthetic
excellent lecturer - and, not least, as a wonderful colleague! We
successful scientific career. His work on the structure and func-
versity of Basel where he got deeply involved in modern NMR
organic molecules, always with the aim of resolving the structure
wish you, Buko, all the best for the years to come.
tion of lipid A became very famous, and it took only some years
techniques and protocols.
faster than the biochemists – mostly successfully.
developed carbohydrate and lipid structural analyses by NMR
that he became one of the few top LPS experts worldwide which
he still is. From November 1983 through March 1984, he joined
RCB said good-bye to Uli by holding a fare-well symposium in
The success of this technique for organic mass analysis in a rather
the laboratory of Prof. Dr. Tetsuo Shiba at the University of Osa-
September 2012, in which many friends, colleagues and co-ope-
high mass range gave rise to questions concerning the mecha-
ka (Japan) for a sabbatical during which he was further trained
rators participated who celebrated his life-time achievement.
nisms of laser ionization. This was then one of the topics for Buko
in analytical and synthetic carbohydrate chemistry. This was the
Still, Uli is around, highly motivated, working on new projects….
Linder‘s habilitation in physics, which he finished in 1993 at the Uni-
begin of a long lasting cooperation and friendship, which con-
We all should count ourselves lucky!
tinued with Prof. Shiba’s successor, Prof. Dr. Shoichi Kusumoto.
21
Farewell
be mass analyzer (LAMMA). This instrument allowed the mass spectrometric analysis of small particles as i.e. bac-
2011
JULES HOFFMANN –
GRANDSEIGNEUR DER WISSENSCHAFT MIT EINEM
FAIBLE FÜR DROSOPHILA UND MARZIPAN
von Prof. Dr. Ulrich Zähringer und Dr. Bettina Brand
JULES ALPHONSE HOFFMANN ERHIELT 2011 GEMEINSAM MIT BRUCE BEUTLER UND
RALPH M. STEINMAN DEN NOBELPREIS FÜR PHYSIOLOGIE ODER MEDIZIN. SCHWERPUNKT SEINER WISSENSCHAFTLICHEN ARBEITEN WAR UND IST DAS HORMONELLE
ABWEHRSYSTEM DER FLIEGE (Drosophila melanogaster). sein AUGENMERK
LIEGT AUF DEN ANTIINFEKTIVA, DIE VON FLIEGEN SYNTHETISIERT UND ZUM TEIL
VON EINEM PROTEIN NAMENS TOLL AKTIVIERT WERDEN. BEIM MENSCHEN EXISTIEREN ÄHNLICHE PROTEINE, DIE TOLL-LIKE RECEPTORS GENANNT WERDEN, UND
EINE WICHTIGE ROLLE BEI DER UNTERSCHEIDUNG DES IMMUNSYSTEMS ZWISCHEN
„SELBST“ UND „NICHT-SELBST“ SPIELEN.
Highlights 2011
23
JULES HOFFMANN ERHIELT FÜR SEINE BAHNBRECHENDEN ARBEITEN UNZÄHLIGE
RENOMMIERTE AUSZEICHNUNGEN UND IST SEIT 2012 MITGLIED DER ACADÉMIE
FRANÇAISE.
Im Februar 2012 folgte Jules A. Hoffmann einer Einladung
allen auf beeindruckende Weise auf „Augenhöhe“ diskutiert
nur ihm gelten kann und konnte. Er kam sofort zum Wesent-
wichtig, die letzten beiden Absagen für die Cluster-Lecture
des Exzellenzcluster ‚Entzündung an Grenzflächen‘ und be-
und er hat das in ebensolcher Weise sein Gegenüber auch
lichen des Forschungsgebietes seines Diskussionspartners,
wieder gut zu machen. Damit hat er auf besonders eindrucks-
suchte die Cluster-Partner in Kiel, Lübeck und Borstel. Das
spüren lassen und ihm damit Respekt gezollt, der eigentlich
war rasch in das Metier eingedrungen und bewegte sich
volle Weise seine Wertschätzung für unsere Arbeit hier im
kompetent und viel wissend auch auf fremdem Terrain. Der
Zentrum gezeigt und bewiesen. Jules Hoffmann hat unserem
Borstel. Daneben führte er zahlreiche Gespräche mit Kolle-
„Grandseigneur der Fliegenforschung“ machte auf uns den
Institut und den dort arbeitenden Forschern großen Respekt
gen und Kolleginnen und nahm an einer Roundtable Dis-
Eindruck größter Bescheidenheit, die sich in seiner Person
und Anerkennung entgegen gebracht, die ich aus seinem
kussion mit Nachwuchswissenschaftlern/innnen des Clusters
mit einer außerordentlich hohen Klugheit paart und dadurch
Munde und in dieser Form nicht erwartet habe. Wir haben
teil. Ganz persönliche Eindrücke dieses Besuchs schildert im
noch stärker und weit imponierender wirkt. Mit einem Nobel-
bei seinem Besuch einen großen Wissenschaftler kennen und
Folgenden Ulrich Zähringer, jetzt im Ruhestand befindlicher
preisträger sprechen zu können ist sicherlich ein Privileg, das
auch bewundern gelernt. Gelernt haben wir auch, dass Be-
Leiter der Immunchemie am FZB.
man so schnell nicht vergisst, insbesondere wenn einem ein
scheidenheit und Klugheit und nicht nur auf Sensation ausge-
Wissenschaftler gegenüber sitzt, der immer noch über eine
legte Wissenschaft erfolgreich sein kann:
Herzstück dieses Besuchs war seine Lecture im Herrenhaus
Stefan Ehlers im Gepräch mit Jules Hoffmann
Wer geglaubt hat einen abgehobenen Nobelpreisträger der
kindliche Neugierde und einen unstillbaren Wissensdurst ver-
Medizin anzutreffen, der mit der Verleihung dieses höchsten
fügt, die wohl beide auch den Grundstock für seinen Erfolg
„Trommle Reveille mit Jugenkraft,
Preises der Wissenschaft den Olymp erfolgreich erklommen
darstellen. Beeindruckt hat mich auch, wie Jules Hoffmann
marschiere trommelnd immer voran,
hat und nun den Rest aller Forscher als „Fußvolk der Wis-
mit diesem Termin in Borstel und seinen Besuch beim Exzel-
das ist die ganze Wissenschaft„
senschaft“ betrachtet, der konnte beim Gespräch mit Jules
lenzcluster umgegangen ist. Er hat – trotz des Streiks des
(Heinrich Heine)
Hoffmann mit seinen eigenen und selbst aufgebauten Vorur-
Flughafen-Bodenpersonals und trotz aller widrigen Umstän-
teilen rasch aufräumen. Stattdessen saß uns ein mit wachen
de – seinen Besuch in Borstel nicht abgesagt, sondern er
Eben nicht – und nicht erst seit dem Besuch von Jules Hoff-
Augen dreinschauender Jules Hoffmann gegenüber, der weit
hat die Mühe auf sich genommen, den frühen Flieger von
mann wissen wir, dass sich „shooting stars“ und Wissen-
davon entfernt war sich in seinem Erfolg überschwänglich
Straßburg nach Hamburg zu nehmen. Offenbar war es ihm
schaft eben doch gegenseitig ausschließen.
bewundern, verehren und feiern zu lassen. Er hat mit uns
2011
Highlights
Werkstattgespräche zur Tuberkulose im
Medizinhistorisches Museum am Universitätsklinikum Hamburg-Eppendorf mit Ulrich
Schaible, Programmdirektor Infektionen.
6. Juni 2011- Einweihung des neuen Laborgebäudes
durch die Kuratorin Dr. Cordelia Andreßen.
Neues BMBF NanoCare Verbundprojekt
‚CarbonBlack‘. Prädiktion humantoxikologischer Wirkung synthetischer Carbon Black
Nanopartikel, Projektleitung Heinz Fehrenbach, Direktor des Programmbereichs Asthma und Allergie.
Farewell Symposium für Ernst Brandt, Borsteler
Wissenschaftler, leidenschaftlicher Chemokinforscher
der ersten Stunde und erfolgreicher Leiter der
Biologischen Chemie.
Year of Science 2011 – Research for Our Health
launched a dialogue which was open for everyone. Leibniz Action Week – featuring:
‘Return of the White Plague’ presented by Stefan Ehlers, Managing Director.
June 6th, 2011 – inauguration of the new laboratory
building by Curator Dr. Cordelia Andreßen.
New NanoCare funding program ‚Carbon Black‘. sponsored by
the Ministry of Education and Science. The joint project ‘Prediction of the human-toxicological effect of synthetic carbon
black nanoparticles’ is led by Heinz Fehrenbach, Director of the Priority Research Area ‘Asthma and Allergy’.
‚Farewell Symposium‘ dedicated to Ernst Brandt,
passionate chemokine scientist of the first hour and
successful head of the research team Biological
Chemistry. Ernst Brandt retired beginning of 2011
but will keep in touch.
The prize for the best thesis awarded by the district
Segeberg went to Kolja Schaale,
Microbial Interface Biology.
Der Preis des Kreises Segeberg für die beste Doktorarbeit
ging 2011 an Kolja Schaale
aus der Forschungsgruppe – ‚Mikrobielle Grenzflächenbiologie‘
Ehrendoktorwürde an Ekkehard Vollmer
Am 03.06.2011 hat die Medizinische und Pharmazeutische Universität Craiova
in Rumänien Ekkehard Vollmer, Leiter der Klinischen und Experimentellen
Pathologie, die Ehrendoktorwürde für seine wertvollen, internationalen
Aktivitäten zur Förderung der Pathologie zuerkannt.
Der Förderverein VEIAP zeichnete 2011 zwei Nachwuchswissenschaftler aus: Andreas Baier
für die beste Diplomarbeit im Bereich Immunbiologie
und Saskia Hellmig für die beste Masterarbeit
im Fachbereich Allergologie.
Tuberculosis-Workshop Discussions
Medical Historical Museum, University
Medical Center, Hamburg, co-chaired
by Ulrich Schaible, Director of the Priority
Research Area Infections.
Ausstellung „MenschMikrobe: Infektionskrankheiten heute“ mit Vortragsreihe des Leibniz Center
Infection zu neuen Erkenntnissen der Infektionsforschung für Jedermann – mit Christoph Lange
(Klinische Infektiologie): Tuberkulose 2011:
Alles unter Kontrolle?
Eva und Klaus Grohe-Preis der berlin-brandenburgischen Akademie
der Wissenschaften wird für herausragende wissenschaftliche Leistungen
auf dem Gebiet der Infektiologie - sowohl in der klinischen Forschung als
auch der Grundlagenforschung – verliehen. 2011 erhielt den mit
20.000 Euro dotierten Preis Stefan Niemann, Leiter der
Molekularen Mykobakteriologie.
Honorary Doctorate awarded to Ekkehard Vollmer.
The University of Craiova, Romania, awarded Ekkehard Vollmer, head of
Clinical and Experimental Pneumology, an honorary doctorate for his
outstanding international activities in promoting pathology.
The friends’ association VEIAP honoured 2011 two
young scientists for their excellent scientific work:
Andreas Baier (Dipl. Biol.), Immunobiology,
and Saskia Hellmig (MSc), Clinical and
Molecular Allergology.
Exhibition “MenschMikrobe: Infectious Diseases
today“ including a series of lectures focusing
on new developments in infection research for
everyone, organized by the Leibniz Center
Infection. Tuberculosis 2012 – is everthing under
control? Presented by Christoph Lange; Clinical
Infectious Diseases.
Eva und Klaus Grohe-Prize of the Berlin-Brandenburg Academy of
Science and Humanities is awarded biennially for outstanding scientific
achievement by German scientists with a doctorate in the area of
infectious diseases – both in clinical research and in basic research. In
2011 the prize was awarded to Stefan Niemann, head of
Molecular Mycobacteriology.
25
HIGHLIGHTS 2011
Wissenschaftsjahr 2011 Forschung für unsere Gesundheit
“Wissenschaft für jedermann“, Leibniz-Aktionswoche im Deutschen Museum München mit dabei:
die Wiederkehr der weißen Pest mit Stefan Ehlers, Zentrumsdirektor.
2012
Highlights
Der Preis des Kreises Segeberg für die beste
The prize for the best thesis 2012 awarded by
Doktorarbeit ging 2012 an Max Koistinen aus
the district Segeberg went to Max Koistinen,
Das EU Konsortium PATHO
der Forschungsgruppe ‚Biophysik‘
NGenTrace
Consortium PATHO
Biophysics.
NGenTrace:
befasst sich mit dem Aufbau eines effektiven
next generation-sequencing for microbial disease
Überwachungs- und Warnsystems mikrobieller
surveillance and early warning systems.
Erkrankungen mittels ‚next generation-sequencing‘.
Coordinated by Stefan Niemann.
Koordination: Stefan Niemann.
Neues Graduiertenkolleg‚ Gene, Umwelt und Entzündung‘ (Kiel-FZB-Lübeck)
Im Fokus steht die Entschlüsselung des Einflusses von Umweltfaktoren auf
New Graduate School‚ (Kiel-FZB-Lübeck) funded by the DFG.
27
The Graduate School will focus on “Genes, Environment and Inflammation”
pathophysiologische Prozesse komplexer, chronischer Erkrankungen.
dissecting the influence of environmental factors on the pathophysiology of
complex and chronic diseases.
Der Förderverein VEIAP zeichnete 2012 Jannike
Blank, Forschungsgruppe‚
Zelluläre Mikrobiologie‘ für ihre hervorragende Masterarbeit aus.
The friends’ association VEIAP honoured 2012 Jannike
Cellular Microbiology, for her excellent scientific work (MSc).
Farewell Symposium für drei Urgesteine der
Helmut Brade,
Ulrich Zähringer und Buko Lindner
Farewell Symposium for three veterans of endotoxin research:
Endotoxinforschung:
Das Internationale Graduiertenkolleg „Immun-
gehen in den Ruhestand. Ohne sie wäre die Struktur, Chemie
regulation der Entzündung bei Allergien und Infektionen“
und Biologie des Endotoxins weiterhin ein Rätsel.
(Lübeck-Kiel-Borstel) wurde positiv begutachtet und wird
2013 seine Arbeit aufnehmen.
Blank,
The International Graduate School Immunregulation of Inflammation in Allergies and Infections’
Helmut Brade, Ulrich Zähringer
and Buko Lindner retired in 2012.
Numerous friends traveled to Borstel
to create a stimulating Farewell-Symposium.
(Lübeck-Kiel-Borstel) was positively evaluated by the DFG
and will start in 2013.
Zwei auf einmal: das FZB ist stolz als Partner des Airway Research
Center North Teil des Deutschen Zentrums für Lungenforschung zu sein. Die thematischen
Translationseinheit „Tuberkulose“, die von Borstel aus koordiniert wird, hat
sich im Rahmen des Deutschen Zentrums für Infektionsforschung das Ziel
Two at a time: we are proud to be a member of the Airway
Research Center North, partner in the German Center for Lung Research. The mission
of the Thematic Translational Unit „Tuberculosis“, module of the German Center for
Infection Research coordinated by the FZB, is to improve TB infection control,
gesetzt, die Kontrolle und Bekämpfung von TB-Infektionen zu verbessern.
Erfolgreiche 2. Förderrunde für den Exzellenzcluster
Cluster of Excellence ‚Inflammation at Interfaces’:
‚Inflammation at Interfaces’: das Phänomen
second funding period for decoding the
Entzündung entschlüsseln.
Ulrich Seydel (Prof. emer.) wurde in Anerkennung seiner wissenschaftlichen Lebensleistung und seines
with a focus on M/XDR-TB.
phenomenon of chronic inflammation.
Susanne Homolka erhielt
Susanne Homolka
den Gertrud-Meissner-Preis der
received the
European Society for Mycobacterio-
Gertrud-Meissner-Prize for her
logy, der Wissenschaftler/innen in
excellent scientific work in
der frühen Phase ihrer Karriere für
mycobacterial research awarded
exzellente Leistungen in der Myco-
Ulrich Seydel recieved the „Honorary Lifetime
by the European Society for
bakteriologie auszeichnet.
Membership“ of the ‚International Endotoxin and Innate
Mycobacteriology.
Engagements in der ‚International Endotoxin and Innate
Immunity Society‘ in recognition of his scientific life-time
Immunity Society‘ der „Honorary Lifetime Membership“
achievement and dedication.
Award auf der Tagung in Tokyo verliehen.
HIGHLIGHTS 2012
Cellular Microbiology
Phagosomes
Mycobacterium
Innate Immunity
Dr. Christian Alexander • Jannike Blank • Dr. Tobias Dallenga • Jacqueline Eich • Anna Christina Geffken
• Nina Grohmann • Kristine Hagens • Nadine Harmel • Dagmar Meyer • Carlotta Ober-Blöbaum • Emmanuel Patin (London School of Hygiene & Tropical Medicine) • Natalja Redinger • Steffi Renk • Dr. Bianca
Schneider • Dr. Yeojun Yun • Dr. Andrea Zelmer (London School of Hygiene & Tropical Medicine)
inflammation
Prof. Dr. Ulrich E. Schaible
Tuberculosis
MISSION
Mueller, A.K., Behrends, J., Hagens, K., Mahlo, J., Schaible,
Stenotrophomonas maltophilia virulence properties: Frank Schmidt,
To analyze host-pathogen interactions in tuberculosis and other lung-associated bacterial infections
on the molecular, cellular and animal model level with a strong focus on the interplay between bacterial virulence properties and host immune responses within the micro-ecosystem of the infected lung.
U.E., Schneider, B.E. Natural transmission of Plasmodium
University Greifswald; Wolfgang Streit, University Hamburg
berghei exacerbates chronic tuberculosis in an experimental
Lysosomal phospholipases in infection: James Shayman, AnnAr-
co-infection model. PLoS One. 2012;7(10):e48110.
bor; Sven Hammerschmidt, University Greifswald
most important findings
controlled in co-infected mice compared to single infected ones,
Herbst, S., Schaible, U.E., Schneider, B.E. 2011 Interferon
29
Mycobacterium (M.) tuberculosis, the causative bacterial agent
which is probably due to differential production of cross-influ-
Gamma Activated Macrophages Kill Mycobacteria by Nitric
Grant support
of tuberculosis (TB), is a facultative intracellular pathogen of ma-
encing cytokines such as interleukin 10 massively triggered by
Oxide Induced Apoptosis. PlosOne 6(5) e19105.
DFG Specific Priority Program 1580 (“Intracellular compartments
crophages. Blocking phagosome maturation is a prerequisite
plasmodium (Mueller et al., 2012). These findings highlight the
for intracellular survival and growth. Interferon gamma (IFN-γ)
importance of future studies in co-infected human populations in
Carroll, P., Schreuder, L.J., Muwanguzi-Karugaba, J., Wiles,
BMBF Pathogen-Host Interactomes (“Infection pathogenomics“)
is a key cytokine for activation of host effector functions of mu-
geographical areas where both infections overlap.
S., Robertson, B.F., Ripoll, J., Ward, T.H., Bancroft, G.J.,
DFG (“Role of lysosomal phospholipases in infection“)
rine macrophages against mycobacteria to overcome phago-
The world-wide TB crisis and the rise of M. tuberculosis strains
Schaible, U.E., Parish, T. 2010 Sensitivie Detection of Gene
Danish Research Council (“Center of Nano-vaccines“)
some maturation block (Ehlers & Schaible, 2013). We identified
resistant to more than one anti-TB drugs calls for an accele-
Expression in Mycobacteria under Replicating and Non-Repli-
Bill-and-Melinda-Gates-Foundation (“Imaging TB“)
an indirect pathway how IFN-γ activated murine macrophages
rated search for novel compounds and more effective pro-
cating Conditions Using Optimized Far-Red Reporters. PlosOne
kill intracellular mycobacteria depending on nitric oxide (NO)
phylactics as the current BCG vaccine. Through the Bill-and-
5 (3): e9823.
driven apoptotic cell death but not autophagy (Herbst et al.,
Melinda-Gates funded “Imaging TB“ consortium lead by UES,
2011). These data further highlight IFN-γ as essential cytokine
sensitive fluorescent and bioluminescent reporter strains of
Zelmer A., Carroll P., Andreu N., Hagens K., Mahlo J., Re-
in protective immunity against TB in mice. The observation that
the TB agent were established (Carroll et al., 2010). These
dinger N., Robertson B.D., Wiles S., Ward T.H., Parish T.,
as places of pathogen-host-interactions”)
an anti-microbial host defense factor, NO, can also indirectly
reporters are combined with in vivo imaging devices, the com-
Ripoll J., Bancroft G.J., Schaible U.E. 2012 A new in vivo mo-
affect host-pathogen interaction has its counterpart in neutrophil
mercial IVIS system as well as a Fluorescent Molecular Tomo-
del to test anti-tuberculosis drugs using fluorescence imaging.
derived reactive oxygen intermediates (ROI). We could show
graphy (FMT) set up custom made through collaboration with
J Antimicrobial Chemotherapy Aug; 67(8):1948-60.
that virulent M. tuberculosis but not attenuated strains lacking
laser physicists from Crete (FORTH). This innovative approach
the virulence associated RD1 region induces ROI in neutrophils.
allows direct imaging of experimental TB in living mice and
internal and external collaborations
In turn, ROI kill these otherwise highly effective phagocytes by
excised lungs infected with those reporter strains in order to
Lipidomics of lysosomal membrane degradation defects: Buko
necrotic cell death and thereby facilitates mycobacterial esca-
monitor infection, i. e. mycobacterial expansion or elimination
Lindner
pe from the neutrophil‘s microbicidal effectors (Corleis et al.,
in response to antibiotic treatment (Zelmer et al. 2012). These
Antimicrobial peptides in tuberculosis: Thomas Gutsmann
2012). As neutrophils represent the predominant mycobacteria-
imaging techniques are currently further employed in vaccine
Mycobacterial glycolipids in phagosome maturation: Otto Holst
infected cell population in lungs of active TB patients, our data
studies against M. tuberculosis and S. pyogenes in a collabo-
M. tuberculosis strain-specific host-pathogen interactions in neu-
indicate that these cells rather contribute to exacerbation of
rative Center for Nanovaccines (CNV) together with the Sta-
trophils: Stefan Niemann
immunopathology than to protective immunity.
tens Serum Institutet, Copenhagen, and funded by the Danish
Phagosome Biology: Norbert Reiling
Recently, we extended the perspective of lung infection re-
government. Those studies are advanced by the multipurpose
Stenotrophomonas maltophilia knock out mutants: Uwe Mamat
search by including the respiratory tract micro-ecology. So far,
usage of nanoparticles for imaging phagocyte and vaccine
Imaging tuberculosis: Tanya Parish, QMUL; Brian Robertson,
the lower respiratory tract was considered sterile but more re-
trafficking but also as vaccine and drug carriers with the ul-
ICL; Gregory Bancroft, LSHTM; Jorge Ripoll, FORTH.
cent studies in human specimen suggested lung commensals.
timate aim to improve both, infection models and anti-(myco)
Imaging vaccine efficacy: Center for Nanovaccines, Peter An-
Using the murine model, we were able to pivotally identify a
bacterial prophylaxis and treatment.
dersen, Staten Serum Institutet, Copenhagen Denmark; Bernd
diverse bacterial lung microflora by isolation, deep sequencing
Kreikemeyer, University Rostock.
and FISH (Yun et al., submitted). The immunological function
selected puBlications
Iron in infection: Robert Hider, Helen Collins, Kings College London
of lung commensal isolates are currently analysed in detail in
Corleis, B., Korbel, D., Wilson, R., Bylund, J., Chee, R.,
Lung microbiome: Kenneth Bruce, Kings College London; John
development, infection and inflammation.
Schaible, U.E. 2012 Escape of Mycobacterium tuberculosis from
Baines, MPI Evolution Biology, Plön; Pamela Ronald, UC Davis.
Cross influences of immune responses against co-infecting pa-
oxidative killing by neutrophils. Cell Microbiol 14(7):1109-1121.
Pathogen-Host Interactomes: Thomas Rudel, Würzburg; Hubert
thogens became evident by our studies on malaria-tuberculosis
Hilbi, Reinhard Haas, Max-von-Pettenkofer Institute Munich, Mi-
co-infections in mice. Whereas M. tuberculosis benefitted from
Ehlers, S., Schaible, U.E. The granuloma in tuberculosis: dy-
chael Hensel, Osnabrück; Dörthe Becher, Greifswald; Monika
a concurrent plasmodium infection, the parasites were better
namics of a host-pathogen collusion. Front Immunol. 2012;3:411.
Hagedorn, BNI
Figure 1: Mycobacterium tuberculosis reporter strain can be imaged in
(I) infected mice. (II) excised lungs, as well as (III) lung tissue sections to
be employed for drug and vaccine efficacy
Priority Research Area Infections • Cellular Microbiology
Phagosome Biology: Albert Haas, University Bonn
MEDICAL AND BIOCHEMICAL MICROBIOLOGY
Lipopolysaccharide
Ute Agge • PD Dr. Sven Müller-Loennies • Christine Schneider • Veronika Susott
Antibody
Carbohydrate Recognition
Lectin
Structure
Prof. Dr. Helmut Brade
complex between toll-like receptor 4, myeloid differentiating factor
Brade L, Heine H, Raina S, Klein G, Di Padova F, Brade H,
Carbohydrate recognition by proteins of the adaptive and innate immune system is pivotal to combat
infectious diseases. Our interest focusses on the structural and functional analyses of antibodies
and lectin binding to lipopolysaccharide (LPS, endotoxin) of Gram-negative bacteria. The goal is the
identification of epitopes to be used for diagnostic and therapeutic purposes and to reveal binding
Mechanisms leading to specificity versus cross-reactivity.
2 and LPS revealed that epitopes of both TLR-4 and mAb WN1 222-
Müller-Loennies S. Immunization with an anti-idiotypic antibody
5 overlap to a large extent. Thus, it is likely that the high affinity of
against the broadly lipopolysaccharide-reactive antibody WN1
the antibody binding prevents the formation of the ternary complex
222-5 induces Escherichia coli R3-core-type specific antibodies
with TLR-4 and consequently immune cell activation.
in rabbits. Innate Immun. 2012, 18(2):279-93.
In continuation of our work on the structural and functional
aspects of LPS, in collaboration with Satish Raina and Gracjana
Makrypidi G, Damme M, Müller-Loennies S, Trusch M,
most important findings
Klein, now at Gdansk University of Technology (Poland), we have
Schmidt B, Schlüter H, Heeren J, Lübke T, Saftig P, Braulke T.
After many years of efforts and in collaboration with our external
investigated the genetic basis of the structural heterogeneity of
Mannose 6 dephosphorylation of lysosomal proteins mediated
partners Stephen Evans from the Univeristy of Victoria (Canada)
E. coli K-12 LPS. Analyses of LPS from isogenic E. coli strains with
by acid phosphatases Acp2 and Acp5. Mol Cell Biol. 2012,
and Paul Kosma from the University of Natural Resources and
nonpolar mutations in the waa locus or overexpression of their
32(4):774-82.
Life Sciences in Vienna (Austria) we have succeeded in resolving
cognate genes revealed that waaZ and waaS are the structural
the structure of the monoclonal antibody (mAb) WN1 222-5 in
genes required for the incorporation of the third 3-deoxy-α-D-
Evans DW, Müller-Loennies S, Brooks Cl, Brade L, Kosma P,
complex with a high affinity ligand at 1.73 Å resolution. This mAb
manno-oct-2-ulosonic acid (Kdo) and rhamnose (Rha) of the
Brade H, Evans SV. Structural insights into parallel strategies
until now is the only antibody able to neutralize and cross-react
inner core, respectively. The incorporation of Rha occurs after
for germline antibody recognition of lipopolysaccharide from
among LPS from E. coli, Salmonella, Citrobacter and Shigella.
the sequential incorporation of the Kdo trisaccharide and this
Chlamydia. Glycobiology. 2011, 21(8):1049-59.
structural heterogeneity was under the overlapping control of the
RpoE σ factor, two-component systems (BasS/R and PhoB/R), and
internal and external collaborations
ppGpp. For the accumulation of a glycoform with three Kdo and
Established collaborations exist with Prof. Dr. S. Evans from
4-amino-4-deoxy-L-arabinose in the lipid A ppGpp was required
the Institute for Biochemistry & Microbiology at the University
in the absence of the RpoE-specific anti-σ factor rseA. A mutant
of Victoria, BC in Canada. Prof. Evans is an internationally
synthesizing tetraacylated lipid A Δ(waaZ lpxLMP) exhibited
recognized expert on X-ray crystal structure analysis. Further
synthetic lethality at 21-23°C pointing to the significance
support comes from Prof. Dr. P. Kosma of the Institute for
of the incorporation of the third Kdo for cell viability at lower
Organic Chemistry at the University for Agricultural Sciences in
temperatures and an influence on membrane function.
Vienna, Austria. Prof. Kosma is an expert in chemical synthesis
We have provided experimental support in the study of cellular
of carbohydrates, has synthesized oligosaccharides which
Figure 2: Conformation and the stabilizing network of hydrogen bonds
mechanisms leading to the dephosphorylation of mannose
could not be obtained by isolation from natural sources for
within the E. coli R2 oligosaccharide bound to mAb WN1 222-5 (pdb
6-phosphate in lysosomal proteins by using an antibody
immunizations, binding studies and structural analyses. Dr.
code 3V0W).
fragment (scFv) specific for mannose 6-phosphate. This work
C.R. MacKenzie from the Institute for Biological Sciences at the
species is not only based on the occurrence of a conserved epitope
was performed with Thomas Braulke from the University Clinic
National Research Council of Canada, ON, Canada, contributes
The structure of the complex (Fig. 1) revealed for the first time
within certain endotoxins but also on the structural properties of the
Hamburg Eppendorf (Germany) which revealed an important
with his expertise in biosensor based binding studies.
a three-dimensional view of a structurally conserved epitope
antibody. The complex between the mAb WN1 222-5 and LPS is
role of acid phosphatases Acp5 acting in concert with Acp2
Work related to the use of a mannose 6-phosphate specific
formed by the inner core region. The conformation of a
almost exclusively formed by interactions with the complementarity
for the complete dephosphorylation of lysosomal proteins.
antibody has been performed with Prof. Dr. T. Braulke from the
complete core oligosaccharide comprising 12 residues could be
determining regions of the heavy chain. The lack of light chain
In collaboration with Dr. Gibbes Johnson, US Food and Drug
Department of Paediatrics at the University Clinic Hamburg
determined. Binding studies using oligosaccharides which were
contribution leaves an unoccupied open space for sugars of
Administration (Bethesda, USA) we have used this novel antibody
Eppendorf in Hamburg (Germany). Further, we support work by
selectively modified by acid degradation revealed a strong
the outer core and the polysaccharide of the O-antigen which
derived scFv in the development of protocols improving the
Dr. G. Johnson, Laboratory of Chemistry, Division of Therapeutic
influence of distant residues on the binding. In spite of the fact
is produced by these bacteria during infections. The generally
preparation of lysosomal enzymes.
Proteins, Center for Drug Evaluation and Research, US Food
that these residues were not directly part of the epitope they
encountered difficulty in obtaining true cross-reactive antibodies
exerted a very significant influence on the epitope formation
against the LPS core may be explained by the predominant use of
selected puBlications
the development of improved protocols for the preparation of
over a long distance within the molecule. A large network of
a different set of germline genes and extensive affinity maturation.
Gomery K, Müller-Loennies S, Brooks Cl, Brade L, Kosma P,
lysosomal enzymes being used as therapeutics
intramolecular hydrogen bonds within the ligand stabilized the
It could also be answered why mAb WN1 222-5 protects against
Di Padova F, Brade H, Evans SV. Antibody WN1 222-5 mimics
bound conformation of the ligand (Fig. 2).
endotoxic activities despite binding to the distant non-toxic part
Toll-like receptor 4 binding in the recognition of LPS. Proc Natl
Grant support
The structural basis of the cross-reactivity towards LPS from different
of the molecule. The comparison to the structure of the ternary
Acad Sci U S A. 2012, 109(51):20877-82.
Our work has not been financed by extramural funding.
Figure 1: The E. coli R2 core oligosaccharide bound to mAb WN1 222-5
(pdb code 3V0W).
and Drug Administration, Bethesda, MD, USA who is aiming at
31
Priority Research Area Infections • MEDICAL
xxxxxxxxxxxxxxxxxxxxx
& BIOCHEM. MICROBIOLOGY
MISSION
Immunochemistry
Structure-Function Relationship of bacterial PAMPs
Dr. Christian Alexander • Birte Buske • Dr. Nicolas Gisch • Nina Grohmann • Heiko KäSSner • Brigitte Kunz • PD Dr. Buko Lindner • Helga Lüthje • Hermann Moll • Bettina Oppermann • Wiebke Schnoor
NMR
Mass Spectrometry
MISSION
Lipidomics and detailed structural analysis of particular
lipopolysaccharide from Capnocytophaga canimorsus reveals
The isolation, purification, and structural characterization of bacterial cell wall components, so
called pathogen /microbial -associated molecular patterns (PAMPs or MAMPs), represents the research
field of the Division of Immunochemistry within the priority area infection. Here we focus on PAMPs
derived from Gram-negative bacteria (e.g. endotoxin (LPS)). Recently the acquired know-how on structural analysis (NMR, MS) has also been extended towards PAMPs from Gram-positive bacteria (e.g. LTA,
WTA, PGN). In this context PAMPs derived from Mycobacteria (M. tuberculosis) differing in their pathogenicity and virulence are investigated as well. Our group is also engaged in the priority area asthma
and allergy in which allergic reactions in the host are investigated. Here we analyze the qualitative
and quantitative (MS-based) influence of membrane lipids (lipidomics) in allergic reactions, this way
covering both priority areas of the Center‘s mission.
lipid species
an unexpected role of the core-oligosaccharide in MD-2 binding.
In recent years, indications for a direct modulation of the
PLOS PATHOGENS 2012 May;8(5):e1002667.
immune system by the lipid composition of cell membranes have
attracted considerable attention. Signaling processes caused
Krauel K, Weber C, Brandt S, Zähringer U, Mamat U,
by infections or allergic reactions are influenced by membrane
Greinacher A, Hammerschmidt S. Platelet factor 4 binding
lipids. To identify these processes on the molecular level in more
to lipid A of Gram-negative bacteria exposes PF4/heparin-like
detail, we successfully established a sensitive lipidomics method
epitopes. BLOOD 2012 Oct;120(16):3345–3352.
based on high resolution HPLC-Fourier-transform MS (and MS/
MS) to identify and quantify the molecular lipid composition of
Schmidt H, Hansen G, Singh S, Hanuszkiewicz A, Lindner
membrane compartments (e.g. phagosomes, signaling platforms,
B, Fukase K, Woodard RW, Holst O, Hilgenfeld R, Mamat
lung epithelial cells) isolated during biological experiments.
U, Mesters JR. Structural and mechanistic analysis of the
most important findings
it resistant against cationic antimicrobial peptides (CAMPs), this
Using this non-targeted approach, we were able to identify
membrane-embedded glycosyltransferase WaaA required for
Capnocytophaga canimorsus has an unusual rough-type
way explaining how this pathogen survives and escapes from
statistically
lipopolysaccharide synthesis. PROCEEDINGS OF THE NATIONAL
LPS structure
the macrophage’s defense in the host.
sphingomyelin content in murine tracheal airway epithelium cells
C. canimorsus is able to provoke fulminant septic syndromes of
Re-evaluation of the structural model for lipoteichoic (LTA)
isolated from healthy mice and mice sensitized and challenged
unknown origin and sometimes of severe outcome in humans,
and wall teichoic acid (WTA) from the lung pathogen
with the relevant antigen house dust mite extract, respectively.
4. Schmidt RR, Pedersen CM, Qiao Y, Zähringer U. Chemical
which were bitten from dogs or cats. We investigated the structure-
Streptococcus pneumoniae
Phosphoinositides (PIPs) seem to play an important regulatory role
synthesis of bacterial lipoteichoic acids: an insight on its biological
significant
differences
in
the
ceramide
and
ACADEMY OF SCIENCES 2012 Apr;109(16):6253–6258.
function relationship of the C. canimorsus LPS (rough-type; LA-
Teichoic acids (pnTA) of the Gram-positive lung pathogen
for the survival of intracellular living bacteria. However, information
significance. ORGANIC & BIOMOLECULAR CHEMISTRY 2011
core) by using preparative HPLC and NMR, which allowed for the
Streptococcus
wall
which of the PIP isomers are involved are so far not available
Apr;9(7):2040–2052.
first time the analysis of a complete LPS molecule in water.
components of this bacterium. They both possess a very complex
due to the lack of appropriate methods. PIP1 and PIP2 isomers
The 1H,13C-HSQC-spectrum (Figure 1) of C. canimorsus rough-
structure with identical repeating units and are highly decorated
cannot be distinguished by MS or by MS/MS nor by HPLC with
5. Gisch N, Buske B, Heine H, Lindner B, Zähringer U. Synthesis
type LPS showed excellent resolved signals, allowing the
with phosphorylcholine (P-Cho). These P-Cho residues function
reasonable detection limits.We have now established a sensitive
of biotinylated muramyl tripeptides with NOD2-stimulating activity.
assignment of all structural and conformational details
as anchors for surface-exposed choline-binding proteins (CBPs),
HPLC-MS method allowing the identification and quantification of
BIOORGANIC & MEDICINAL CHEMISTRY 2011 Jun;21(11):3362–3366.
within the glycolipid – in water, without the use of expensive
which serve as immune protective molecules and are involved in
the three PIP1 isomers after deacylation (Figure 2).
C-labelling. After completion of the structural NMR analysis,
essential physiological functions of S. pneumoniae (e. g. bacterial
internal and external collaborations
adhesion to host cells). In cooperation with the group of Prof. Dr. S.
Inhouse
Hammerschmidt (University of Greifswald) we investigated various
Heine, Ulmer, Holst, Mamat, Reiling, Niemann, Gutsmann, Schromm,
O-deacylated pneumococcal lipoteichoic acid (pnLTA) preparations
Grassl, Schaible, A. Petersen, F. Petersen, Fehrenbach, Jappe
from nonencapsulated pneumococcal strains by NMR spectroscopy
National
and ESI-MS spectrometry in an unprecedented detailed manner.
Schmidt (Konstanz), Hammerschmidt (Greifswald), Bekeredjian-
The data obtained allowed us to (re)define the former model
Ding (Heidelberg), Bufe (Bochum), Scheel (Halle), Rosenstiel,
in several aspects and to correct inconsistencies existing in the
Röder (Kiel), König, Hübner (Lübeck), Hakenbeck (Kaiserslautern),
literature. The new structure now represents the only one being in
Flieger (RKI Wernigeroda), Heuer (RKI Berlin), Hilbi (München)
complete agreement with all known structural, biosynthetical, and
International
immunological data. Furthermore, we could show for the first time
Grzesiek, Cornelis (Basel), Knirel, Kondakova (Moscow), Raina,
that the terminal sugar residues of the pnLTA [Forssman disaccharide;
Klein (Gdansk), Hermoso (Madrid), Pedersen (Kopenhagen),
13
Figure 1: 1H,13C-HSQC NMR (700 MHz) from HPLC-purified LA-core of C.
canimorsus recorded in 1.5 % (w/w) DHPC-d40 at 325 K. Regions of the
anomeric, ring, and fatty acid signals are surrounded by dotted lines.
pneumoniae
represent
essential
cell
α-D-GalpNAc-(1→3)-β-D-GalpNAc-(1→)] can be heterogeneous
with respect to its degree of phosphorylcholine substitution in the
O-6-positions of both GalpNAc-(1→) residues. Furthermore, we
33
Figure 2: Extracted ion chromatogram showing deacylated PIP1 standards (top) and phagosomal PIPs isolated from macrophages (bottom).
Corsaro (Naples)
Grant support
started to analyze the structure of pnWTA, aiming to proof the so far
selected puBlications
Exzellenzcluster Inflammation at Interfaces (IRN G TP2; 2008-
the endotoxin remains intact and suitable for further biological
unknown linkage of the pnWTA to the peptidoglycan and providing
Ittig S, Lindner B, Stenta M, Manfredi P, Zdorovenko E,
2012), SFB TR 22 (Z01; 2005-2012), SPP 1580 (2011-2014), DFG
investigations. The unusual LPS structure of C. canimorsus makes
the basis for a deeper knowledge of the biosynthesis of pnTAs.
Knirel YA, dal Peraro M, Cornelis GR, Zähringer U. The
(Za-149/6-1; 2008-2011)
Priority Research Area Infections • Immunochemistry
Prof. Dr. Ulrich Zähringer
Ursula Schombel • Simone Thomsen • Michael Weinkauf • Nicole Zehethofer
Biophysics
Host defense peptides
Lipid membrane
Glycolipid
Prof. Dr. Klaus Brandenburg • Sabine Dabelstein • Niels Denkert • Sabrina Groth • Nina Hahlbrock
Christine Hamann • Dr. Lena Heinbockel • Yani Kaconis • Annika Kopp • Christian Nehls • Dr. Leyre Palacios • Michaela Ramhold • Kerstin Stephan
Signal transduction
Lipid-peptide interactions
MISSION
force microscopy we then detected a loading rate dependent
Hornef, Medical School Hannover; T. Schürholz, Medical School
The structure of lipid membranes and their interaction with peptides and proteins is in the focus of
the Division of Biophysics. To characterize the molecular function of Host Defence Peptides (HDPs) and
of various bacterial pathogenicity factors we use biological as well as biophysical techniques. To
simplify the complex structure of lipid membranes we use isolated membranes, purified membranes, individual lipid species and synthetic lipids to mimic the natural membranes of bacteria and human cells.
specificity shift. At higher loading rates higher forces were need
Aachen; B. Schittek, University of Tübingen; A. Haas, University
to break the bond between FimH and the glucoside. Two effects
of Bonn; M. Winterhalter, Jacobs University, Bremen; G. Marti-
are important: (i) FimH and the carbohydrates form catch bonds
nez de Tejada, Universidad de Navarra, Pamplon; Sunil David,
and (ii) the dissoziation rate as well as the transition state dis-
University of Kansas, Lawrence, USA
tance depends on the specific carbohydrate. The different transition state distance of the glucoside leads to the counterintuitive
Grant support
most important findings
of symmetric TDM bilayers and was also able to stabilize lipid
effect that despite a weak dissoziation rate higher force were
German Ministry BMBF (grant 01GU0824; ‘Therapy of infectious
Anti-Sepsis Project
membranes. Domain formation could also be observed by high
needed to break the bond between the glucoside and FimH at
diseases with special regard to bacterial sepsis’)
One of our main projects is the development of effective antiin-
resolution atomic force microscopy on life mycobacteria. When
high loading rates. This result was confirmed by surface acoustic
Deutsche Forschungsgemeinschaft (GU 568/4-1, ‘Biophysical
fective peptides directed against severe infections such as the
incorporated into lipid membranes the long alky chains of TDM
wave measurements using high flow conditions. The loading rate
investigations into the interactions between antimicrobial pep-
bacterial sepsis. The development has led so far to a polypep-
were folded, possibly promoting H-bonds and van-der-Waals in-
dependent-specificity is a new aspect and can have high impact
tides of the epithelial defense and microbial cell envelopes’;
tide lead structure Pep19-2.5, also called ‘Aspidasept’. The ability
teractions with neighboring molecules. In addition, experiments
on an understanding of bacterial infections.
AN 301/5-1 ‚Significance of cellular surface factors for the selec-
of compound Pep19-2.5 to suppress the inflammation reaction in
with HDPs revealed the peculiar role of TDM in mycobacterial
tive cytotoxicity of antimicrobial peptides (AMPs) against cancer
vitro was investigated by adding the pathogenicity factors to hu-
HDP resistance. Results showed that the active human hBD-3-l
selected puBlications
cells‘; Schwerpunktprogramm 1580 “Intracellular compartments
man mononuclear cells (MNC) or macrophages and monitoring
was able to permeabilize TDM containing bilayers by binding to
Andrä J, Goldmann T, Ernst CM, Peschel A, Gutsmann T. Mul-
as places of pathogen-host-interactions”)
their response with respect to the release of proinflammatory
lipid areas and formation of peptide domains with a very high
tiple peptide resistance factor (MPRF)-mediated resistance of
Leibniz-Graduiertenschule (Adhesion of Plasmodium-infected
cytokines (tumor-necrosis factor (TNFα, interleukin-6, interferonγ),
local peptide concentration. Interestingly, the hBD-3-l activity was
staphylococcus aureus against antimicrobial peptides coincides
erythrocytes to the human endothelial receptors CD36, and P-
prostaglandins (PGE2), and reactive oxygen species (ROS). As
nullified, when the membranes had been treated with another
with a modulated peptide interaction with artificial membranes
selectin - Characterization of binding properties)
main bacterial pathogenicity factor of Gram-negatives, lipopoly-
human HDP called LL32 before. LL32 was by itself unable to
comprising lysyl-phosphatidylglycerol. JOURNAL OF BIOLOGI-
Else Kröner-Fresenius-Stiftung (Biophysikalische Charakterisierung
saccharides were taken. We could show that the LPS-induced
permeabilize TDM containing bilayers but caused a partial van-
CAL CHEMISTRY 2011 May 27;286(21):18692-700.
der Interaktion von Endotoxinen mit nicht-steroiden antientzündli-
TNFα production could be considerably inhibited by Pep19-2.5,
ishing of the TDM domains.
and in a similar way this was true also for interferon (IFNγ) and
In a subsequent hBD-3-l addition this peptide was now unable
Brauser A, Schroeder I, Gutsmann T, Cosentino C, Moroni A,
reactive oxygen species (ROS). It should be emphasized that the
to act on TDM-free membrane areas, as TDM and lipids were
Hansen U-P, Winterhalter M. Modulation of enrofloxacin bind-
peptides does not inhibit the cytokine production induced by
mixed. hBD-3-l activity was impaired either by direct interaction
ing in OmpF by Mg2+ as revealed by the analysis of fast flicker-
synthetic Pam3CSK4, which is a TLR2 agonist and is a shortened
with TDM molecules or by TDM mediated stabilization of the lipid
ing single-porin current. THE JOURNAL OF GENERAL PHYSIOL-
variant of MALP-2. We have also tested in vitro activity of Pep19-
bilayer. An LL32 triggered inhibition of hBD-3-l activity was not
OGY 2012 Jul;140(1):69–82.
2.5 in combination with antibiotics. The next step was the in vivo
only found in model membranes but was also demonstrated us-
chen Wirkstoffen und deren Modifikation durch antimikrobielle
activity against endotoxicity and bacteremia in a mouse model.
ing Mycobacterium tuberculosis bacilli. For a further investigation
Schuerholz T, Dömming S, Hornef M, Dupont A, Kowalski I,
Almost 100 % of the mice were protected 5 days after LPS chal-
of TDM, small-angle X-ray scattering experiments were carried
Kaconis Y, Heinbockel L, Andrä J, Garidel P, Gutsmann T,
lenge when the treatment was administered immediately after
out. By this, the TDM conformation in solution, in planar lipid bi-
David S, Sánchez-Gómez S, de Tejada GM, Brandenburg
the challenge. When mice received the peptide with a delay of
layers and planar lipid multilayers was examined. Furthermore,
K. Bacterial cell wall compounds as promising targets of antimi-
30 min with respect to the LPS, the protection was very significant
HDP influence on the membrane organization was investigated.
crobial agents II. Immunological and clinical aspects. CURRENT
Peptide)
Figure 1: Atomic force microscopic images of a life BCG bacterium
before and after addition of the antimicrobial peptide LL32 (in collaboration with N. Reiling and JPK Instruments).
DRUG TARGETS 2012 Aug;13(9):1131-7.
during the first 48 h but later the majority of the animals died.
Loading rate determines lectin specificity (in collaboration
Figure 2: LPS-induced
Interaction between mycobacteria and HDPs
with T. Lindhorst)
internal and external collaborations
As the mycobacterial outer membrane (MOM) plays an impor-
Lectins are carbohydrate binding proteins with high specificity.
Internal cooperation:
tissue. Determination of TNF-α
tant role in mycobacterial virulence and resistance to antibiotics,
We have performed binding studies with the α-D-mannoside-
Norbert Reiling, Ulrich Schaible, Niels Röckendorf and Andreas
expression levels from human
this membrane was reconstituted as planar bilayers and GUVs.
specific lectin FimH, abundant on the tips of bacterial type 1
Frey, Otto Holst, Torsten Goldmann and Ekkehard Vollmer,
lung tissue after incubation
Membrane models contained the particular important mycobac-
fimbriae. Bacteria, e.g. Escherichia coli, need the fimbriae to
Andra Schromm
terial glycolipid trehalose dimycolate (TDM), whose structural
adhere on surfaces. At static equilibrium conditions, compared
External cooperation:
properties and interactions with AMPs were investigated. Results
to an α-D-glucoside, α-D-mannosides were recognized with high
R. Schmitz-Streit, University of Kiel; T. Lindhorst, University of Kiel;
showed that TDM in lipid matrices gathered into stable domains
specificity. In single binding force measurements utilizing atomic
M. Leippe, University of Kiel; C. Hübner, University of Lübeck; M.
inflammation of human lung
with S. enterica LPS (in collaboration with T. Goldmann).
35
Priority Research Area Infections • Biophysics
Prof. Dr. Thomas Gutsmann
Immunobiophysics
Macrophage activation and regulation
Surfactant Protein
Irina von Cube • Sabrina Groth • Susanne Keese • Franziska Kopp • Yvonne Siebken • Mona Shokouhi
Sonja Winkler
Membrane Biophysics
Immune Modulators
PD Dr. Andra Schromm
MISSION
Torsten Goldmann, Division of Clinical and Experimental
The division of IMMUNOBIOPHYSICS 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.
Pathology
Thomas Gutsmann, Division of Biophysics
37
Buko Lindner, Division of Immunochemistry
Cordula Stamme, Division of Cellular Pneumology
Ulrich Zähringer, Division of Immunochemistry
External collaborations:
Manfred Roessle, HASYLAB c/o DESY, Hamburg
most important findings
procedure. Biological analysis of LPS fractions derived from E.
Michael Steinert, Technical University, Braunschweig
Bacterial membrane lipids as regulators of host innate
coli demonstrated that the rough (R)- and smooth (S)-fractions
Andreas Tholey, Proteomics Unit, Christian-Albrechts-University
immunity
were highly active in inducing TNFα in human macrophages
Kiel
Lipopolysaccharides of the outer membrane of bacterial cell
in the presence of human serum, however, R- fractions
Karl-Heinz Wiesmüller, University Tübingen, EMC microcollections
walls are potent stimulators of the innate immune system. The
were by a factor of 10-100 more active than the S-fraction.
EISAI Research Institute, Andover, USA
Figure 2: Binding of LBP to HEK293 cells.
bacterial cell wall contains a variety of chemically different
Under serum-free conditions, the natural LPS preparation as
LPS molecules with different immune stimulatory capacity.
well as the R-fractions showed dose-dependent activation of
cell membranes. Cell-associated LBP appears to be involved
Aiming at characterizing the potency and mechanisms of
macrophages, whereas the S-fraction did not induce TNFα even
in the activation of immune responses to LPS, a function
biological activity of the individual molecular species we
at high concentrations. The dose-response of the natural LPS
that has not been widely recognized so far. We have set up
Grant support
have isolated S- and R-form LPS-species from natural LPS
preparation resembled that of the R-fractions which indicates
to express and purify recombinant human LBP to study the
DFG SCHR 621/3-1: ”Immunregulation in der Lunge durch pul-
preparations by a newly established procedure based on
that short-chain (R- and SR-form) LPS dominate the innate immune
structure, function and dynamics of the membrane-associated
monale Collectine”
preparative DOC-slab-PAGE. The isolated LPS fractions were
response of human macrophages to natural LPS preparations
LBP. We have studied the binding of fluorophore-labeled LBP to
Leibniz Graduate School “Modell systems for infectious diseas-
analyzed by mass spectrometry which demonstrated that no
in-vitro. The biological activity to S-fraction LPS could be restored
human macrophages and HEK293 cell lines expressing the LPS
es“: “Investigations into the cellular function of the acute-phase
significant chemical modifications were introduced by the
under serum-free conditions by addition of recombinant
receptor proteins. These experiments demonstrate a fast and
protein LBP in antimicrobial immunity”
lipopolysaccharide-binding protein (LBP), whereas sCD14 was
dose-dependent binding of LBP to the cytoplasmic membrane
not able to confer activity of S-fraction LPS, indicating a crucial
followed by rapid uptake in macrophages, whereas HEK293
role of LBP for the recognition of S-LPS by human macrophages.
cells showed a delayed intracellular localization. The cellular
Our data demonstrate that R- and S-form molecular species
binding of LBP was strongly enhanced by the presence of
from natural LPS preparations differ in the initiation of signaling
LPS, suggesting a role of membrane bound LBP as part of the
pathways. The new methodology for preparative fractionation
cellular LPS-signaling complex. The mechanisms of membrane
of LPS preparations provides the basis for a detailed analysis
interaction and the topology of LBP localization are currently
of the molecular mechanism and the selectivity for signaling
investigated in reconstituted membranes models and host cells.
pathways by individual molecular species of LPS.
Investigations into the cellular function of the acute-phase
selected puBlication
protein LBP in antimicrobial immunity
Pupo E, Lindner B, Brade H, Schromm AB. Intact rough- and
Immune recognition of microbial associated molecules via
smooth-form lipopolysaccharides from Escherichia coli sepa-
the toll-like receptor system is a central part of the innate
rated by preparative gel electrophoresis exhibit differential bio-
immune defense against infections. The acute-phase protein
logical activity in human macrophages. FEBS J 2012, in press.
lipopolysaccharide-binding protein (LBP) is a key player in the
recognition of bacterial membrane lipids and orchestrates the
internal and external collaborations
cellular immune response to a variety of cell wall components
Internal collaborations:
Figure 1: Biological activity of LPS fractions isolated from natural LPS
such as LPS and lipopeptides. LBP does not only have a
Helmut Brade, Division of Immunochemistry and Biochemical
preparations (Printed abstract from Pupo et al., FEBS J 2012)
strong affinity for bacterial lipids but also interacts with host
Microbiology
Elder Pupo, Bilthoven, Netherlands
Priority Research Area Infections • Immunobiophysics
Uwe Mamat, Division of Structural Biochemistry
Molecular Inflammation Medicine
granuloma necrosis
Tuberculosis
Martina Ackermann • Silvia Maass • Dr. Sven Malm • Dr. Judith Petersen • Christina Trabandt
Dr. Kerstin Walter
TBornotTB network
experimental therapy
Prof. Dr. Stefan Ehlers
DZIF Thematic Translational Unit
MISSION
were suggested as a major resistance mechanism in clinical
Herzmann C, Lange C, Schaberg T, Ernst M, Ehlers S;
The division makes use of the mouse model of aerogenic tuberculosis for identifying host and pathogen determinants of TB disease progression and for improving therapeutic strategies.
isolates. To directly analyze the impact of individual embB306
TBornotTB Consortium. Immunological evidence of incipient
mutations on EMB resistance, allelic exchange experiments
pulmonary tuberculosis. J Infect Dis. 2012; Nov 15;206(10):1630-1.
39
immunity enhancing strategies that merely mimic the natural immune
to generate embB306 mutants of M. tuberculosis H37Rv. The
Maertzdorf J, Weiner J 3rd, Mollenkopf HJ; TBornotTB
A granuloma is defined as an inflammatory mononuclear cell
response directed against M. tuberculosis infection can overcome
level of EMB resistance conferred by particular mutations
Network, Bauer T, Prasse A, Müller-Quernheim J, Kaufmann
infiltrate that, while capable of limiting growth of Mycobacterium
pulmonary tuberculosis in the adult population. Juxtaposition of
was measured in vitro and in vivo after EMB therapy by daily
SH. Common patterns and disease-related signatures in
tuberculosis, also provides a survival niche from which the bacteria
molecular pathology and immunology with microbial physiology
gavage in a mouse model of aerogenic tuberculosis. The wild-
tuberculosis and sarcoidosis. Proc Natl Acad Sci U S A. 2012
may disseminate. The tuberculosis lesion is highly dynamic and
and the use of novel imaging approaches afford an integrative view
type embB306 ATG codon was replaced by embB306 ATC,
May; 15;109(20):7853-8.
shaped by both, immune response elements and the pathogen
of the granuloma’s contribution to the life cycle of M. tuberculosis.
ATA, or GTG, respectively. All of the obtained embB306 mutants
(see Figure 1). In the granuloma, M. tuberculosis switches to a non-
Our laboratory analyzes the input of innate and adaptive immunity
exhibited a 2- to 4-fold increase in EMB MIC compared to the
Thye T, Niemann S, Walter K, Homolka S, Intemann CD,
replicating but energy-generating life style whose detailed molecular
in granuloma biogenesis, with a focus on the co-evolutionary forces
wild-type H37Rv. In vivo, the one selected embB306 GTG mutant
Chinbuah MA, Enimil A, Gyapong J, Osei I, Owusu-Dabo E,
characterization can identify novel targets for chemotherapy. To secure
that redirect immune responses also to the benefit of the pathogen,
required a higher dose of ethambutol to restrict its growth in
Rüsch-Gerdes S, Horstmann RD, Ehlers S, Meyer CG. Variant
transmission to a new host, M. tuberculosis has evolved to drive T cell
i.e., its survival, propagation, and transmission.
the lung compared to wild-type H37Rv. These experiments
G57E of mannose binding lectin associated with protection
demonstrate that embB306 point mutations enhance the EMB
against tuberculosis caused by Mycobacterium africanum but not
cavities to facilitate expectoration of bacilli. From an evolutionary
Individually and combined, Toll-like receptors (TLR)-2, -4, -9,
MIC in vitro to a moderate, but significant extent, and reduce
by M. tuberculosis. PLoS One. 2011;6(6):e20908. doi: 10.1371.
perspective it is therefore questionable whether vaccination and
nucleotide oligomerization domain (NOD) 2 and NALP3 contribute
the efficacy of EMB treatment in the animal model. We propose
to the Mycobacterium tuberculosis (Mtb)-induced innate immune
that conventional EMB susceptibility testing, in combination with
response only to a limited extent, particularly in terms of inducing
embB306 genotyping, may guide dose adjustment to avoid
tuberculosis embB codon 306 mutations confer moderately
antibacterial protection and granuloma formation in vivo. A
clinical treatment failure in these low-level resistant strains.
increased resistance to ethambutol in vitro and in vivo. Antimicrob
immunity to the point that necrotizing granulomas leak into bronchial
singular essential sensory component of this initial response
Agents Chemother. 2011 Jun;55(6):2891-6.
has not been discovered yet. Trehalose-6,6’-dimycolate (TDM),
Researchers in the Division contribute to the Thematic
a well known mycobacterial cell wall glycolipid, is believed to
Translational Unit Tuberculosis, a programmatic research area
internal and external collaborations
be involved in these early inflammatory processes after Mtb
within the newly founded German Center for Infection Research
TB Center Borstel; Christian G. Meyer, Rolf Horstmann, BNI,
infection. Only recently the macrophage inducible C-type lectin
(DZIF). The DZIF Mouse Test Station for anti-TB therapies will use
Hamburg; Ulrich Maus, Experimental Pneumology, MHH
(Mincle) was demonstrated as an essential receptor for TDM.
existing 200 sq.m. BSL3 capacity and in vivo imaging methodology
Hannover; Franz-Christoph Bange, Medical Microbiology,
However, not much is known about the sensing capacity of
to further develop an aerogenic TB model in transgenic mice
MHH
Mincle during infection with live mycobacteria. To determine the
that closely mimicks human pathology, monitor treatment efficacy
University of Birmingham/AL (USA); Peter Sander, Medical
significance of Mincle during tuberculosis (TB), we analyzed the
using bioluminescent and fluorescent Mtb strains, test natural
Microbiology, University of Zurich (CH); Steffen Stenger, Medical
outcome of Mtb infection in Mincle-deficient mice. Whereas in
compounds, chemicals and already existing drugs in dosing and
Microbiology, University of Ulm; Sergej Nedospasov, Deutsches
the absence of Mincle macrophages did not respond to TDM,
combination regimes for antimycobacterial efficacy in the mouse
Rheumaforschungszentrum, Berlin
Mincle-deficient mice were capable of mounting an efficient
lung, and test the efficacy of lentivirally transduced bone-marrow
granulomatous and protective immune response after low
derived stem cells for enhancing innate immunity in the lung.
Grant support
and high dose infections with Mtb. Mutant mice generated a
The Division was discontinued in July 2012 following the appointment
DFG/BMBF: Cluster of Excellence Inflammation at Interfaces,
normal T helper (TH) 1 and TH17 immune response followed
of the head of the Division as full-time Managing Director of the
EXC 306 (2007-2012); BMBF: Network “Pulmonary Tuberculosis
by the induction of efficient macrophage effector mechanisms
Research Center Borstel. All research activities will be continued and
– Host and Pathogen Determinants of Resistance and Disease
and control of mycobacterial growth identical to wildtype mice.
all cooperations with other institutions and in national consortia will
Progression (TBornotTB)”, (2007-2014); BMBF: Network “Short
From our results we conclude that absence of the innate receptor
be honored by other Divisions of the Priority Area Infections.
course chemotherapy against tuberculosis” (2007-2012); DFG
Mincle can be fully compensated for in vivo in terms of sensing
Figure 1: M. tuberculosis (Mtb) elicits a local inflammatory infiltrate
which may give rise to (i) protective immunity, (ii) balanced inflammation (i.e., control of Mtb growth with little tissue damage), or (iii)
endobronchial transmission following granuloma necrosis.
Plinke C, Walter K, Aly S, Ehlers S, Niemann S. Mycobacterium
Mtb and mounting a protective inflammatory immune response.
Hannover;
Michael
Niederweis,
Microbiology,
Eh 101/10-1: The role of serine proteases in experimental tu-
selected puBlications
berculosis (2011-2014); BMBF: DZIF Thematic Translational Unit
Ehlers S, Schaible U. The granuloma in tuberculosis: dynamics
Tuberculosis (2011-2015)
Ethambutol (EMB) is a major component of the first-line therapy
of a host-pathogen collusion. Front. Imm. 2013; doi: 10.3389/
of tuberculosis. Mutations in codon 306 of embB (embB306)
fimmu.2012.00411.
Priority Research Area Infections • Molecular Inflammation Medicine
were performed in the Division of Molecular Mycobacteriology
most important findings
Infection Immunology
Alternatively activated
macrophages
Tuberculosis
Chagas
Dr. Christoph Hölscher
Mahin Abad Dar • Martina Ackermann • Dr. Jochen Behrends • Julia Böhme • Dr. Hanna Erdmann
Dr. Lisa Heitmann • Alexandra Hölscher • Kristian Holz • Melanie Janssen • Gabriele Röver • Erik Schmok
Jan Christian Sodenkamp • Anna Stubbe • Kerstin Traxel • Johanna Volz • Dr. Kerstin Walter
TH17
MISSION
elimination of intracellular pathogens in macrophages and that
mechanisms against Trypanosoma cruzi by trapping parasites
We want to understand the cytokine-mediated regulation of protection, persistence and pathology in
order to dissect protective and pathology-promoting mechanisms associated with the inflammatory
response during chronic infectious diseases such as tuberculosis (TB) and Chagas disease.
an IL-17A-driven phagocytosis prolonged the containment in
in the endolysosomal compartment. Immunobiology. 2012 Oct
the endosomal/lysosomal compartment. This IL-17A-dependent
26. doi:pii: S0171-2985(12)00515-3. 10.1016/j.imbio.2012.10.005.
mechanism represents a novel function of IL-17A, trapping
[Epub ahead of print].
41
most important findings
remodelling we hypothesized that arginase activity is involved
enhancing the exposure time to antimicrobial mechanisms of
internal and external collaborations
TB infection starts with the inhalation of infectious Mycobacterium
in mediating granuloma necrosis. To prove this hypothesis,
the macrophage. Whereas the TH17 cytokine IL-17A is required
Ehlers S, Division of Molecular Inflammation Medicine, Research
tuberculosis (Mtb) into the lung and the uptake of bacteria by
we induced arginase-1 independently of IL-13 in wildtype
for the immune defence against Mtb and T. cruzi infection
Center Borstel; Schaible U, Division of Cellular Microbiology,
resident alveolar macrophages. This host-cell/bacilli interaction
mice by blockade of the endogenous arginase inhibitor
we could show that IL-22 is dispensable for host protection.
Research Center Borstel; Thye T, Meyer CG, and Horstmann
leads to the induction and secretion of a variety of cytokines
L-hydroxyarginine with N6-(iminoethyl)-L-lysine (L-NIL). L-NIL
Surprisingly, we did not find IL-17A/IL-22 co-producing TH17 cells.
R, Department of Molecular Medicine, Bernhard-Nocht-
and chemokines. Via this process, other immune cells are
treatment of Mtb-infected wildtype mice resulted in a strong
Instead, IL-22 was solely expressed in TH22 cells or produced
Institute for Tropical Medicine, Hamburg, Germany; Jacobs T,
recruited to the site of infection leading to the formation of
arginase expression and precipitated a similar pathology of
by IFNγ-secreting TH1 cells. Hence, these IL-22-expressing TH1
Bernhard-Nocht-Institute for Tropical Medicine, Department
granulomas in order to contain mycobacterial growth. However,
granuloma necrosis as observed in IL-13tg mice So far, we have
cells might also represent a transition state of TH22 to TH1
of Immunology, Hamburg, Germany; Lang R, Molecular
in some patients the infection reactivates and these granulomas
shown that IL-13/IL-4Rα-induced arginase-1 expression is directly
cells. Taken together, our results suggest that IL-17A but not IL-
Medicine, University Hospital Erlangen, Germany; Rose-John
may necrotize. Mechanisms leading to granuloma necrosis,
involved in TB-associated tissue pathology.
22 contributes to host protection by stimulating anti-microbial
S, Department of Biochemistry, Christian-Albrechts-University
mechanisms in macrophages.
Kiel, Germany; Müller U, University of Leipzig, Germany;
the hallmark of human TB pathology, are not understood. In
Brombacher F, International Centre for Genetic Engineering and
a human study group, a structural variant of the interleukin (IL)-
Biotechnology, University of Cape Town, South Africa.
4 receptor-alpha (IL-4Rα), the common receptor chain for IL-4
In summary, our studies so far revealed that TH2 and TH17
and IL-13, was found to be significantly associated with the
immune
cavity size of human granulomas. Because centrally necrotizing
macrophage effector responses after infection. Currently,
Grant support
granulomas are usually absent in wildtype mice, it has been
we are trying to disclose macrophage-specific downstream
BMBF;
difficult to experimentally assess the involvement of IL-4/
mechanisms during infectious diseases in vivo by the generation
Desing trypanozider Wirkstoffe, Teilprojekt B“
and analysis of conditional gene-deficient mice.
BMBF:
IL-13-mediated mechanisms in the tissue pathology of TB. To
responses
regulate
or
stimulate
anti-microbial
KMU-innovativ-5:Verbundprojekt:
Nation-wide
Collaborative
„Struktur-basiertes
Grant:
“Pulmonary
overcome this obstacle, we established an experimental model
Figure 2: Induction of arginase activity in Mtb-infected wildtype
in which IL-13-overexpressing (tg) mice infected with Mtb develop
mice leads to granuloma necrosis. NOS2 activity was blocked by
selected puBlications
and disease progression.” Workpackage E (Animal models).
centrally necrotizing granulomas with multinucleated giant cells,
treating C57BL/6 50 days after aerosol infection with 100 CFU Mtb with
Heßmann M, Rausch A, Rückerl D, Adam PS, Simon M,
DFG; Cluster of Excellence “Inflammation at Interfaces”:
Gilfillan S, Colonna M, Ehlers S, Hölscher C. DAP10 contributes
Integrative Research Network F “Cytokine signalling via gp
to CD8+ T cell-mediated cytotoxic effector mechanisms during
130”, project 4 “Infection driven inflammation models”.
Mycobacterium tuberculosis infection. IMMUNOBIOL 2011; 216:
Universität zu Lübeck; FSP „Autoimmunität“: Projekt A1 „Die
639-647.
Bedeutung einer TH17 Immunantwort für die Pathogenese der
a hypoxic rim and a perinecrotic fibrosis with an adjacent zone
of lipid-rich, acid-fast bacilli-containing foamy macrophages,
L-NIL (black symbols). Infected control mice were left untreated (white
symbols). Left panel: arginase activity in lung homogenates. Right
panel: collagen staining of a necrotic lung granuloma.
Mostly based on in vitro studies, T helper (TH)17 cells have been
Tuberculosis - host and pathogen determinants of resistance
Epidermolysis bullosa acquisita“
characterized by a lineage-specific production of IL-17A and IL22. However, the discovery of IL-17A/interferon-gamma (IFNγ)
Sodenkamp J, Behrends J, Förster I, Müller W, Ehlers S,
Universität zu Lübeck; FSP „Modulation von Infektion und
co-producing cells in vivo has recently indicated a plasticity
Hölscher C. gp130 on macrophages/granulocytes modulates
Allergie“: Projekt B3 „ Untersuchung der optimalen Zytokin-
between TH17 and TH1 cells. A TH17 immune response is strongly
inflammation during experimental tuberculosis. EUR J CELL BIOL
vermittelten T und B Zell-Antworten nach der Impfung gegen
involved in tissue inflammation and has been associated with
2011; 90: 505-514.
several chronic and autoimmune inflammatory diseases but the
Mycobacterium tuberculosis“
DFG; GRK 1727: „Modulation von Autoimmunität“: Projekt A4
main function of TH17 cells appears to be the clearance of
Sodenkamp J, Waetzig GH, Scheller J, Seegert D, Grötzinger
“Modulation of experimental autoimmune encephalitis by the
pathogens, which are not appropriately affected by TH1 cells.
J, Rose-John S, Ehlers S, Hölscher C. Therapeutic targeting
regulatory T cell-derived cytokine interleukin-35”
thus strongly resembling the pathology in human post-primary
Nevertheless, the protective mechanisms mediated by these
of interleukin-6 trans-signaling does not affect the outcome of
TB. Importantly, granuloma necrosis in Mtb-infected IL-13tg
cells in vivo are not well understood. Despite strong TH1 immune
experimental tuberculosis. IMMUNOBIOL 2012 217: 996-1004.
mice is associated with the induction of arginase-1-expressing
responses, IL-17A-deficeint mice were susceptible to infection
macrophages. Because arginase-1 has been implicated in
with Mtb or Trypanosoma cruzi, the causative agent of Chagas
Erdmann H, Roßnagel C, Böhme J, Iwakura Y, Jacobs T,
susceptibility to infection with intracellular pathogens and tissue
disease. In vitro experiments revealed that IL-17A mediates
Schaible UE, Hölscher C. IL-17A promotes macrophage effector
Figure 1: Model of granuloma necrosis in Mtb-infected IL-13tg mice
Priority Research Area Infections • Infection Immunology
pathogens in intracellular effector compartments thereby
Microbial Interface Biology
Mycobacterium tuberculosis
Dr. Christine Steinhäuser • Julius Brandenburg • Dennis Linke • Lisa Niwinski • Katrin Seeger • Svenja
phagosome isolation
Goldenbaum
pathogen variability
Wnt signaling in inflammation
in vitro drug testing
MISSION
Wenzel UA, Bank E, Florian C, Förster S, Zimara N, Stein-
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
(i) the “novel” functional role of Wnt signaling in inflammation, (ii) the molecular characterization
of the M. tuberculosis phagosome, (iii) the pathobiological consequences of the genetic diversity of
the M. tuberculosis complex strains and last but not least (iv) the testing of novel anti-mycobacterial
agents in primary macrophages.
acker J, Klinger M, Reiling N, Ritter U, van Zandbergen G.
Leishmania major parasite stage-dependent host cell invasion
and immune evasion. FASEB JOURNAL 2012 Jan;26(1):29-39.
Klug K, Ehlers S, Uhlig S, Reiling N. Mitogen-activated protein
kinases p38 and ERK1/2 regulated control of Mycobacterium
avium replication in primary murine macrophages is indepen-
most important findings
tions, as we recently demonstrated the suppression of TNF-alpha
dent of tumor necrosis factor-α and interleukin-10. INNATE IM-
Wnt signaling in macrophages: Augmenting and inhibiting
expression in mycobacteria-infected macrophages by Wnt3a or
MUNITY 2011 Oct;17(5):470-85.
mycobacteria-induced inflammatory responses
inhibition of GSK3beta, respectively. Thus, Wnt-induced signals
Wnt signaling is well known to be an ancient and highly con-
can have both, driving and inhibitory effects on inflammatory pro-
served principle among metazoan species, which executes es-
cesses. Likewise, inflammatory signaling pathways interfere with
sential functions in embryogenesis and tissue homeostasis. In
Wnt-induced signals in a synergistic or antagonistic fashion.
Uciechowski P, Imhoff H, Lange C, Meyer CG, Browne EN,
Kirsten DK, Schröder AK, Schaaf B, Al-Lahham A, Reinert RR,
Figure 2: Scheme of the novel magnetic phagosome isolation protocol.
Reiling N, Haase H, Hatzmann A, Fleischer D, Heussen N,
terium (M.) avium, M. tuberculosis and Listeria monocytogenes and
Kleines M, Rink L. Susceptibility to tuberculosis is associated
cancer and degenerative diseases. In mice and men 19 ligands,
New access to the habitat of intracellular pathogens: Lipid-
isolated magnetic bacteria-containing phagosomes from primary
with TLR1 polymorphisms resulting in a lack of TLR1 cell surface
secreted palmitoylated glycoproteins of the Wnt family, and ten
Labeling Facilitates a Novel Magnetic Isolation Procedure
cells using a strong magnetic field in a novel free-flow system.
expression. J LEUKOC BIOL. 2011 Aug;90(2):377-88.
seven-transmembrane receptors of the Frizzled (Fzd) family as well
to Characterize Pathogen-Containing Phagosomes.
Magnetic labeling of M. tuberculosis did not affect the virulence
as several co- or alternative receptors are known. We and others
Pathogenic microorganisms have developed various strategies to
characteristics of the bacteria during infection experiments. Bio-
Schaale K, Neumann J, Schneider D, Ehlers S, Reiling N.
have recently identified a regulatory role for components of the
avoid detection and elimination by the immune system. Many, like
chemical analyses of the magnetic phagosome-containing frac-
Wnt signaling in macrophages: Augmenting and inhibiting my-
Wnt signaling network to be operative at the interface between in-
Legionella, Brucella, Ehrlichia, and Mycobacterium spp., effectively
tions provided evidence of an enhanced presence of bacterial
cobacteria-induced inflammatory responses. EUROPEAN JOUR-
nate and adaptive immunity: In essence, Wnts can exert both, pro-
manipulate the normal progression of their phagosomal compart-
antigens and a differential distribution of proteins involved in the
NAL OF CELL BIOLOGY. 2011 Jun-Jul; 90(6-7):553-9.
and anti-inflammatory functions on macrophages and other cells
ment and prevent it from fusing with or maturing into an active
endocytic pathway over time as well as cytokine-dependent chang-
of the immune system. Wnt5a was the first member of the Wnt
lysosomal organelle. To identify pathogen-specific virulence and
es in the phagosomal protein composition. The newly developed
internal and external collaborations
family that was described to be induced in multiple inflammatory
persistence mechanisms it is indispensable to define the molecular
method (Fig.2) represents a useful approach to characterize and
• S Niemann, T Gutsmann, B Lindner, N Gisch, U Schaible, G
disease settings such as rheumatoid arthritis, tuberculosis, sepsis,
composition of phagosomes containing viable infectious agents.
compare pathogen-containing compartments, in order to identify
Graßl, K Brandenburg, C Herzmann, C Hölscher, C Lange, S
psoriasis and atherosclerosis and very recently also in obesity
We developed a novel, lipid-based procedure to magnetically la-
microbial and host cell targets for novel anti-infective strategies.
and asthma. In 2006 our group described Wnt5a and its receptor
bel the surface of bacteria and visualized the label by scanning
The outcome of tuberculosis is determined by both the virulence
• S Schütze, A Tholey, University of Kiel; J Rupp, University of
Fzd5 to be induced in human macrophages challenged with M.
and transmission electron microscopy (SEM, TEM; Fig. 1). We per-
of the infecting M. tuberculosis complex (MTBC) strain and the kill-
Lübeck; K-H Wiesmüller, EMC microcollections, Tübingen; E
tuberculosis or conserved bacterial structures. We could assign a
formed infection experiments with magnetically labeled Mycobac-
ing capacity of the host macrophage. Based on our observation
Krause, FMP, Berlin; G van Zandbergen, Paul Ehrlich Institute,
pro-inflammatory function to Wnt5a, a finding later also reported
that MTBC strains show a differential ability of the strains to grow
Langen; T Pukrop, University of Göttingen; I Rosenkrands,
by other groups. Unlike Wnt5a, Wnt3a has been referred to as a
in human macrophages, we are currently examining the impact
Statens Serum Institut, Copenhagen, DK; L Rink, S Uhlig, Uni-
“classical” or “canonical” member of the Wnt family characterized
of pathogen variability and host cell signaling variability on the
versity of Aachen; P Walther, University of Ulm.
by the induction of the Wnt/beta-Catenin signaling pathway. In
molecular composition of the MTC strain harboring phagosomes.
the adult, dysregulation of Wnt signaling is often associated with
Rüsch-Gerdes, U Zähringer.
Grant support
an inflammatory context, activation of the Wnt/beta-Catenin pathway has been associated with a reduced migration of monocytes
selected puBlications
and lately with the induction of tolerance in dendritic cells. A key
Steinhäuser C, Heigl U, Tchikov V, Schwarz J, Gutsmann T,
1 ”The influence of pathogen and host cell variability on the
player in Wnt/beta-Catenin signaling is the Glycogen Synthase
Seeger K, Brandenburg J, Fritsch J, Schroeder J, Wiesmül-
molecular composition of phagosomes harbouring different
Kinase 3beta (GSK3beta), which is also a well known regulator of
ler KH, Rosenkrands I, Walther P, Pott J, Krause E, Ehlers S,
NF-kappaB dependent gene transcription. Its inhibition, as it hap-
Schneider-Brachert W, Schütze S, Reiling N. Lipid-Labeling
pens during activation of Wnt/beta-Catenin signaling, was shown
Figure 1: Scanning electron microscopy of magnetically labeled M.
43
avium. Merged images of the secondary electron image and the
Facilitates a Novel Magnetic Isolation Procedure to Character-
to suppress pro-inflammatory cytokine expression in different ex-
backscattered electron image (pink staining, overlay). In cooperation with
ize Pathogen Containing Phagosomes. TRAFFIC 2012 Dec 11.
perimental settings. This holds true also in mycobacterial infec-
P. Walther, University of Ulm. White arrows indicate the magnetic label.
doi: 10.1111/tra.12031 [Epub ahead of print].
• DFG Re1228/4-1 and DFG Priority Program 1580, Re1228/5-
strains of the Mycobacterium tuberculosis complex”
• DFG Cluster of Excellence 306 ”Inflammation at Interfaces”,
Research Area Z.
• Bundesministerium für Bildung und Forschung (BMBF) Grant
01KI0784,” TB or not TB”, WP D.
Priority Research Area Infections • Microbial Interface Biology
PD Dr. Norbert Reiling
Models of Inflammation
inflammation
Salmonella
Janin Braun • Anne-Kathrin Claes • Yvonne de Graaf • Fabian Müller • Dorothee Schultz • Dr. Natalie
Steck
fibrosis
Prof. Dr. Guntram Grassl
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).
Grassl GA, Faustmann M, Gill N, Zbytnuik L, Merkens H, So
L, Rossi FM, McNagny KM, Finlay BB. CD34 mediates intestinal inflammation in Salmonella-infected mice. Cell Microbiol.
Grassl GA, Valdez Y, Bergstrom KS, Vallance BA, Finlay
BB. Chronic enteric salmonella infection in mice leads to severe and persistent intestinal fibrosis. Gastroenterology. 2008
Mar;134(3):768-80. doi: 10.1053/j.gastro.2007.12.043. Epub 2008
Jan 9.
internal and external collaborations
most important findings
exacerbated inflammation was most likely due to TLR2 / Nod2
INTERNAL: Norbert Reiling: Wnt signalling in gut and lung in-
Role of NOD-like receptors in Salmonella-triggered inflam-
crosstalk.
flammation, Ulrich Zähringer, Holger Heine: TLR4 and NOD2
mation.
crosstalk, Helmut Haas, Gabi Schramm: Schistosoma-Salmonel-
NOD2: NOD-like receptors (NLRs) are pattern recognition recep-
NLRP6: Nucleotide-binding domain and Leucin-rich Repeat-con-
la co-infections.
tors for bacterial ligands. The NLR NOD2 is a cytoplasmic re-
taining Protein 6 (NLRP6) is part of the NLRP6-inflammasome.
EXTERNAL: John Baines, MPI Plön & CAU Kiel: Gut microbiota
ceptor for muramyl-dipeptide (a part of the bacterial cell wall).
Its formation leads to Caspase-1 activation, which processes
Mutations of the NOD-like receptor NOD2 are important risk
pro-IL-1ß and pro-IL-18 to their active forms. Recently, it has been
factors for developing Crohn’s disease, characterised by chro-
shown that NLRP6 contributes to maintaining a healthy gut mi-
nic intestinal inflammation. It is believed that these mutations of
crobiota and that NLRP6-deficient mice develop more severe
NOD2 lead to a loss-of-function phenotype of the receptor. Ho-
DSS-induced colitis. Upon infection, both NLRP6-deficient and
wever, how these mutations result in un-controlled inflammation
wildtype mice developed strong inflammation of the cecum.
B4galnt2 expression. Although intestinal S.T. colonization was
Diehl, Skirball Institute, New York, USA and Brett Finlay, UBC
in the gut is not well understood. Using NOD2-/- and NOD2+/+
However, we found no significant differences in histopathology
enhanced in B4galnt2-deficient mice, they developed signi-
Vancouver, Canada: Th17 response in gut fibrosis
control mice, we showed that the response to wildtype Salmo-
of the cecum, nor in intestinal colonization between wildtype
ficantly less pathology in the cecum compared to wild type
in fibrosis; Philip Rosenstiel, Simone Lipinski, CAU Kiel: InflamFigure 2: Glycosylation in the mouse intestine.
DBA lectin staining reveals B4galnt2-specific glycosylation patterns on
the surface of epithelial cells and in goblet cell of the mouse intestine.
Green: DBA-lectin (N-Glycans), blue: DAPI (nuclei).
mation in NLR deficient mice; Jürgen Brinckmann, University of
Lübeck: Extracellular matrix composition in fibrosis; Ohad GalMor: Molecular and cellular characterization of Salmonella
enterica serovar Infantis pathogenicity; Dan Littman, Gretchen
nella was comparable. In contrast, infection with the attenuated
and NLRP6-deficient animals. Survival curves revealed that NL-
mice one day post-infection. Cecal inflammation in B4galnt2-
Grant support
Salmonella mutant Salmonella Typhimurium ΔmsbB led to an
RP6-deficient mice were more susceptible to Salmonella infec-
deficient was associated with lower numbers of CD68 and
DFG: Excellence Cluster 306 “Inflammation at Interfaces”:
increased inflammation in NOD2-/- mice compared to controls
tion. Bone marrow derived macrophages from NLRP6-deficient
CD3 positive cells and less expression of MCP-1 and IL-6 com-
Mouse models of inflammation
(Figure 1). In vitro transfection of HEK cells showed that the
mice produced more TNF-a, but less IL-1ß upon infection with
pared to wild type mice. Furthermore, changes in the inte-
The German Israeli Foundation for Scientific Researcht and De-
S. Typhimurium. These results demonstrate that NLRP6 is dis-
stinal microbiota correlated significantly with development of
velopment (G.I.F): “Molecular and cellular characterization of
pensable for Salmonella-triggered intestinal inflammation but
inflammation in a B4galnt2 genotype-dependent manner. In
Salmonella enterica serovar Infantis pathogenicity”
that it plays an important role in the systemic response to this
summary, our results show that variation in intestinal glycosyla-
pathogen.
tion patterns alters susceptibility to diseases such as infectious
gastroenteritis.
Intestinal Glycosyltransferases in Salmonella Infections.
mice. Streptomycin-pretreated C57BL/6 and Nod2-/- mice were orally
infected with S. Typhimurium DmsbB for 2 days. H&E staining of infected ceca sections. Original magnification: 100x. L: lumen; M: mucosa;
SM: submucosa.
Glycans on mucosal surfaces play an important role in host-
selected puBlications
microbe interactions. We previously demonstrated that intesti-
Mansson LE, Montero M, Zarepour M, Bergstrom KS, Ma
nal epithelial B4galnt2 expression influences the composition
C, Huang T, Man C, Grassl GA, Vallance BA. MyD88 signa-
of the intestinal microbiota in mice. To determine whether
ling promotes both mucosal homeostatic and fibrotic respon-
B4galnt2 glycans (Figure 2) influence host susceptibility to
ses during Salmonella-induced colitis. Am J Physiol Gastroin-
enteric pathogens, we investigated Salmonella Typhimurium
test Liver Physiol. 2012 Aug 1;303(3):G311-23. doi: 10.1152/
(S.T.) induced gastroenteritis in the presence and absence of
ajpgi.00038.2012. Epub 2012 Jun 7.
Priority Research Area Infections • Models of Inflammation
2010 Nov;12(11):1562-75. doi: 10.1111/j.1462-5822.2010.01488.x.
Using in vivo and in vitro infection models, we try to understand:
i.how enteric pathogens – especially Salmonella enterica - 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
iii.the mechanisms that lead to the development, propagation and resolution of intestinal inflammation and fibrosis
Figure 1: Salmonella induces exacerbated inflammation in Nod2-/-
45
VETERINARY INFECTION BIOLOGY AND IMMUNOLOGY
Tick-Borne Diseases
Co-Infection
Prof. Dr. Jabbar S. Ahmed • Prof. Dr. Ulrike Seitzer • Dr. Mohamed Bakheit • Dr. Stefanie Renneker • Monika
Mackiewicz • Bogdan Oltean • Junlong Liu • Susan Tschitschmann • Sara Mohammed • Doreen Beyer
Birgit Kullmann • Britta Petersen
Tick Allergy
host-pathogen-interaction
theileria
MISSION
of Theileria infected cells from apoptosis and enhance parasite
DAAD funded strategic academic partnerships, DFG funded
The focus of our research activities lies in the host-vector-pathogen interactions. In particular, we
deal with the immunological and molecular biological aspects of the piroplasmosis of small and large farm animals. Second, we deal with the immunopathogenesis of allergy and inflammation, caused
by tick molecules. Furthermore, we are working on co-infection studies of Theileria with M. bovis and
we are involved in M. bovis monitoring in Schleswig-Holstein. Last but not least, we are committed to
the dissemination of knowledge and the capacity building in emerging and developing countries.
proliferation and stage progression. We could show that T.
joint research projects and the Leibniz Graduate School (Model
annulata has two isoforms of HSP90, which were characterized
Systems for Infectious Diseases), the division of Veterinary Infec-
and recombinant expressed. Primary data has shown that
tion Biology and Immunology is linked to numerous research
inhibition of HSP90 in T. annulata induces translocation of p53
institutions at national and international (Europe, Asia, Africa
to the host cell nucleus and subsequent apoptosis of host cells.
and Latin America and Australia) levels.
Allergenic
Grant support
and
immunomodulatory
potential
of
tick
most important findings
differentiation.
molecules
External funding has been raised by the division of Veterinary
Cell division and parasite phosphorylation
Cysteine proteases (CPs) are involved in differentiation of various
The capacity of tick bite to elicit a hypersensitivity reaction was
Infection Biology and – Immunology from EU, DFG and DAAD.
Phosphorylation events play an essential mechanism in cellular
life cycle stages. On the other hand, cysteine protease inhibitors
thoroughly investigated by the means of a human basophil
processes, for example during cell division. Considering that
(ICPs) have been suggested to play a role during differentiation
activation test. The acquired results suggest that tick lysates are
the division of T. annulata schizonts goes along with the division
by fine-tuning the effect of CPs. We identified a Theileria gene
able to induce the basophil activation provided that the patients
of the host cell, it was of interest to find out if phosphorylation
with characteristics of ICP. This is transcribed in the schizonts
were bitten at least twice. In an attempt to characterize the
of schizont proteins and the interaction with host kinases may
regardless of the degree of attenuation. It could be confirmed
tick molecules responsible for basophil activation, tick lsyates
be involved.
that the soluble protein is secreted to the host cells cytoplasm.
were further tested for their IgE reactivity in the serum of bitten
In proteolysis assays, recombinant ICP significantly inhibited
patients. The lysates showed a strong IgE binding capacity
We could show that the schizont interacts via its T. annulata
cathepsins-L, with only minimum enzyme residual activity. Further
using western blot but not in ELISA. In addition, we successfully
Surface Protein (TaSP) with host cell CDK1 in situ and the
characterization of recombinant T. annulata ICP has shown
expressed and purified an Ixodes ricinus Der-p2 allergen-like
schizont surface shows a strong phosphorylation of the epitope
autoproteolytic activity. The conditions for this autoproteolysis
protein, which neither activate the basophiles nor reacted with
phospho(p)Thr-Pro in a cell cycle dependent manner. It could
were not determined; however, it did not interfere with the
IgE antibodies.
be shown that TaSP contains a conserved CDK1 recognition
ability of the ICP domain positioned at the intact C-terminus to
motif [pTPTK] and serves as a substrate for CDK1, shown by the
inhibit cathepsin-L. Ongoing experiments focus on testing ICP
specific phosphorylation of Thr131 by CDK1 in vitro. Functional
functionality against parasite endogenous CPs and host cell
cDNA and the his-tagged protein was expressed in E. coli. The
CDK1 seems to be a key factor for the proliferation of T. annulata-
caspaces, key players in the apoptotic processes taking place
production of TNF-α and IL-10 by LPS-stimulated peripheral
infected cells since CDK1 inhibition prevents proliferation and
during differentiation.
blood monouclear cells were down regulated by recombinant
Salp15-like gene was amplified from I. ricinus salivary gland
Salp15-like protein. Salp15-like protein also shows inhibitory
results in strong morphological changes of the host cell as well
as the parasite, indicating that the parasite can sense stress
In another set of experiments, we could confirm that the potential
function on the production of IL-2 by CD4+ cells. These results
factors and induces an alternative rescue program.
of T. annulata to undergo differentiation is reduced through
showed Salp15-like protein have the functions to interfere with
passaging, possibly attenuation. We could further confirm
some inflammatory cytokines production.
The phosphorylation of TaSP by CDK1 at this specific site
that the ability of a highly passaged/attenuated parasite to
overlaps with a conserved docking site for Polo-Like Kinase 1
form merozoites could be regained after treatment of the
(PLK1), an important cell cycle regulating kinase, which is known
infected cells with a histone deacetylase inhibitor. Thus, histone
selected puBlications
to interact with the schizont surface during mitosis and therefore
modification that is supposedly taking place upon prolonged
Salih DA, Ali AM, Liu Z, Bakheit MA, Taha KM, El Imam AH,
TaSP can be presented very likely as the direct interacting
culturing of the parasite does indeed lead to changes in the
Kullmann B, El Hussein AM, Ahmed JS, Seitzer U (2012).
partner of PLK1, which was unknown up to now.
biological properties of the parasite and leading to a reduced
Development of a loop-mediated isothermal amplification
ability of the parasite to differentiate. In order to evaluate
method for detection of Theileria lestoquardi. Parasitol Res 110(2):
Parasite attenuation and differentiation:
parasite specific histone modulators, we managed to identify
533-538
The differentiation of Theileria schizont is a vital in vivo life
and characterize a novel Theileria Sir2 histone deacetylase with
cycle step, by which formation of merozoites, subsequent
possible involvement in reduced potential of the attenuated
Internal and external collaborations
infection of invertebrate hosts and continuity of survival is
parasite to undergo differentiation.
Effective cooperation and exchange of experience has been
ensured. It is suggested that the parasite becomes attenuated
during culturing, which is associated with reduction/loss of
organized at national, regional and international level.
It is hypothesized that HSP90 plays a major role in protection
Through the intensive participation in EU funded joint projects,
47
Priority Research Area Infections • VET. INFECTION BIOLOGY & IMMUNOLO GY
Prof. Dr. Jabbar S. Ahmed
Cellular Pneumology
Lung innate immunity
Beate Höschler • Linda Lang • Dr. Alexandra Lange • Dr. Christina Moulakakis • Dr. Vicky Sender
Rab GTPases
soluble C-type lectins
endolysosomal trafficking
MISSION
Within preliminary work we established FCS in living cells
ONE, 2013;8(3):e59896. doi: 10.1371/journal.pone.0059896.
The human pulmonary soluble C-type lectins surfactant protein (SP)-A and SP-D are pattern recognition
receptors of lung innate immunity with important functions in lung immune homeostasis in vivo. Our
objective is to understand lung-specific innate imprinting by SP-A and SP-D to Gram-negative bacteria
and M. tuberculosis in order to develop lectin-based antimicrobials and immunotherapeutics.
(Fig. 2). The data show early differences of p65/RelA diffusion
Epub 2013 Mar 25.
parameters in A549 human lung epithelial cells, used as a
model cell system to reflect NF-κB activity in primary immune
Sender V, Moulakakis C, Stamme C. Pulmonary surfactant
cells upon pathogen recognition.
protein A enhances endolysosomal trafficking in alveolar
macrophages through regulation of Rab7. JOURNAL OF
most important findings
SP-A-enhanced GTPases are functionally active as determined by
SP-A mediates lung immune responses partially via its direct
increased interaction of Rab7 with its downstream effector Rab7
effects on alveolar macrophages (AM), the main resident
interacting lysosomal protein (RILP) and enhanced maturation
internal and external collaborations
leukocytes exposed to antigens. SP-A modulates the AM
of cathepsin-D, a function of Rab7b. In AM and RAW264.7
Internal: PD Dr. N. Reiling (Microbial Interface Biology),
threshold of lipopolysaccharide (LPS) activity towards an anti-
macrophages, the SP-A-enhanced lysosomal delivery of GFP-E.
Dr. C. Steinhäuser (Microbial Interface Biology), Prof. Dr. G.
inflammatory phenotype both in vitro and in vivo. LPS responses
coli is abolished by Rab7 small interfering (si)RNA transfection.
Grassl (Models of Inflammation), Prof. Dr. O. Holst (Structural
are tightly regulated via distinct pathways including subcellular
Rab7 blocking peptides antagonize SP-A-rescued lysosomal
Biochemistry).
TLR4 localization and thus ligand sensing. The cytosolic scaffold
delivery of GFP-E.coli in AM from SP-A -/- mice. Activation of Rab7,
National: Prof. Dr. C. Hübner, Dr. M. Knop (Dept. of Physics,
and signaling protein β-arrestin 2 acts as negative regulator of
but not Rab7b, by SP-A depends on the PI3K/Akt/protein kinase
Univ. Lübeck), Prof. Dr. U. Costabel/Dr. F. Bonella (Ruhrlandklinik,
LPS-induced TLR4 activation. We found that SP-A does neither
C (PKC) ζ signal transduction pathway in AM and RAW264.7
Univ. Essen).
increase TLR4 abundancy nor co-localizes with TLR4 in primary
macrophages. The data demonstrate a novel role for SP-A in
International: Prof. Dr. S. Hawgood (Univ of California, San
AM. SP-A significantly reduces the LPS-induced co-localization
modulating endolysosomal trafficking via Rab7 in primary AM
Francisco, USA).
of TLR4 with the early endosome antigen (EEA) 1 by promoting
and define biochemical pathways involved.
IMMUNOLOGY. 2011 Feb; 186(4):2397-411.
Grant support
the co-localization of TLR4 with the post-Golgi compartment
marker Vti1b in freshly isolated AM from rats and wild-type
Since the predominant route of transmission of M. tuberculosis
Work in this group was supported by the Deutsche
(WT) mice, but not in β-arrestin 2-/- AM (Fig. 1). Compared to
is via aerosols, the first cells to encounter M. tuberculosis are
Forschungsgemeinschaft (Grant 609/1-4 and 609/2-1 to C S;
-/-
WT mice pulmonary LPS-induced TNFα release in β-arrestin 2
resident alveolar macrophages (AMφ) whose unique phenotype
Grant 1999/2-1 to C M).
mice is accelerated and enhanced and exogenous SP-A fails
is, in turn, determined by the local intraalveolar environment.
to inhibit both lung LPS-induced TNF-α release and TLR4/EEA1
The initial step of mycobacteria/ AMφ interaction is a central
positioning. SP-A, but not LPS, enhances β-arrestin 2 protein
element of TB pathogenesis and is thus assumed to be decisive
expression in a time-dependent manner in primary rat AM. The
for the outcome of TB infection. As a transient activation
constitutive expression of β-arrestin 2 in AM from SP-A -/- mice is
of NF-κB during the initial stage of infection is required for
significantly reduced compared to SP-A+/+ mice and is rescued
mycobacterial killing, the success of M. tuberculosis in causing
by SP-A. Prolonged endosome retention of LPS-induced TLR4
TB is intimately related to its ability to evade this host defense
in AM from SP-A mice is restored by exogenous SP-A, and
pathway. The project investigates the in vivo dynamics of NF-
is antagonized by β-arrestin 2 blocking peptides. LPS induces
κB/IκB-α proteins in AMφ and its subversion by mycobacteria
β-arrestin 2/TLR4 association in primary AM which is further
at early stages of infection. Constitutively, SP-A and SP-D can
enhanced by SP-A. The data demonstrate that SP-A modulates
alter the macrophages’ activation threshold of the NF-κB/IκB-α
-/-
LPS-induced TLR4 trafficking and signaling in vitro and in vivo
pathway that is supposed to be subverted by M. tuberculosis /
engaging β-arrestin 2.
cell wall glycolipids. In this project, functional proteomics studies
within living macrophages will focus on in vivo protein-protein
Enhanced phagocytosis and endocytosis are key functional
interaction analysis of basal and induced NF-κB activity by
Figure 1: SP-A fails to inhibit the LPS-induced co-loclization of TLR4/EEA1
consequences of AM/SP-A interaction suggesting a SP-A-
fluorescence correlation spectroscopy (FCS), dual color cross
in β-arrestin 2-/- AM
mediated modulation of small Rab GTPases that are pivotal
correlation spectroscopy (FCCS) and in vivo single-molecule
membrane organizers in both processes. We found that SP-A
FRET. The direct and spatially resolved measurement of protein-
specifically and transiently enhances the protein expression of
protein interaction in living cells in real time by these techniques
Sender V, Lang L, Stamme C. Surfactant protein-A modulates
Figure 2: Fluorescence correlation plot and fit function for Pam3CSK4-
endogenous Rab7 and Rab7b in primary AM from rats and mice.
will dissect pathogen- from host-directed signalling pathways.
LPS-induced TLR4 localization and signaling via β-arrestin 2. PLOS
stimulated A549-p65-GFP siRNA knockdown cells
selected puBlications
49
Priority Research Area Infections • Cellular Pneumology
Prof. Dr. Cordula Stamme
Molecular Mycobacteriology
Tuberculosis
molecular epidemiology
Patrick Beckert • Dr. Silke Feuerriegel • Dr. Lisa Heitmann • Dr. Susanne Homolka • Dr. Thomas Kohl
Justina Krawczyk • Anja Lüdemann • Silvia Maass • Dr. Sven Malm • Matthias Merker • Dr. Judith Petersen
Dr. Andreas Roetzer • Tanja Struwe-Sonnenschein • Dr. Barbara Tizzano • Tanja Ubben • Julia Zallet
evolution
Mycobacterium tuberculosis complex
population structure
MISSION
tuberculosis typing methods in a four-year study in Schleswig-Holstein,
Main 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. The link between the
fascinating genomic variability of the pathogen and its association with antibiotic resistance, hostpathogen interaction and virulence represents a highly challenging research topic.
Northern Germany. J Clin Microbiol, 2011, 49(12):4173-8.
Comas I, Borrell S, Roetzer A, Rose G, Malla B, Kato-Maeda
M, Galagan J, Niemann S, Gagneux S. Whole-genome
sequencing of rifampicin-resistant Mycobacterium tuberculosis
strains identifies compensatory mutations in RNA polymerase
genes. Nat Genet, 2012, 44(1):106-10. doi: 10.1038/ng.1038.
most important findings
Epidemiology of TB. Standardized typing
(PA-824). Using NGS, we could identify a novel polymorphism (rplC
51
Figure 1: Clustering numbers of IS6110 RFLP and MIRU-VNTR typing.
Out of 277 isolates, 198 isolates remained unclustered, whereas 79 iso-
Beckert P, Hillemann D, Kohl TA, Kalinowski J, Richter E,
of bacteria and
T460C) that is the major resistance mechanism for the reserve
easy identification of clones exhibiting increased local or global
drug linezolid in clinical isolates. While the data demonstrate
transmissions rates are essential for epidemiological surveillance
that particular SNPs are excellent markers for drug resistance of
and disease control. For MTBC isolates, multi-locus variable
clinical isolates, it also appears to be important to consider the
number tandem repeat analysis (MLVA) targeting mycobacterial
influence of natural genetic diversity in clinical isolates on the
interspersed repetitive units (MIRU) has been internationally
performance of molecular drug susceptibility testing and treatment
Stefan Niemann. Eva and Klaus Grohe Prize of the Berlin-
adopted as the new standard typing method. However, precise
efficacy. Existing phylogenetic variants render some groups of
Brandenburg Academy of Sciences and Humanities 2011.
data on its performance in different settings are not yet available.
the MTBC intrinsically resistant against one or multiple antibiotics.
Susanne Homolka. Gertrud Meissner Award of the European
Therefore, we compared the performance of 24-loci-MIRU-
Moreover, neutral or silent changes within genes responsible for
Society of Mycobacteriology 2012.
lates were clustered. The latter could be subdivided into the following
three groups: clustered by RFLP typing, clustered by MIRU typing, and
Niemann S, Feuerriegel S. rplC T460C Identified as a Dominant
clustered by both (Roetzer et al. JCM 2011 49(12):4173-8).
Mutation in Linezolid-Resistant Mycobacterium tuberculosis
Strains. Antimicrob Agents Chemother, 2012, 56(5):2743-5.
VNTR typing and the Gold Standard IS6110-DNA-fingerprint for
drug resistance can cause false-positive results with hybridization-
discrimination of MTBC isolates in a longitudinal study in Schleswig-
based assays, which have been recently introduced to replace
internal and external collaborations
Holstein. We found an overall high degree of concordance (93 %)
slower phenotypic methods.
Borstel TB Center
between both methodes (Fig. 1). However, our data revealed an
A major step towards a better understanding of the factors involved
D. Harmsen, Univ. Münster; J. Kalinowski Univ. Bielefeld; U. Nübell,
overall higher resolution power of IS6110 RFLP typing, esp. for
in microevolution and fitness of multidrug resistant (MDR) strains has
RKI; S. Gagneux, Swiss TPH, Basel; E. Aleksic, Burnet Institute,
strains of the Beijing lineage. As all typing methods used so far
been made by the identification of compensatory mutations in RNA
Melbourne; K. Kremer, D. van Soolingen, RIVM, Bilthoven; K.
interrogate only a small part of the genome and are not reflecting
polymerase genes. By combining whole genome sequencing and
the full diversity among clinical isolates, we worked towards the
PCR-sequencing of large collections of clinical MDR isolates, we
Figure 1: Mutations identified after genome sequencing of experimen-
Lille; T. Wirth, Muséum Natl. d’Histoire Naturelle, Paris.
establishment of whole genome based molecular epidemiology
could identify mutations in RNA polymerase genes rpoA and rpoC
tally evolved strains (circle) or paired clinical isolates (triangles) are
German Center for Infection Research: 26 partner institutions
for MTBC. In 2012, next generation genome sequencing (NGS)
(Fig. 2) as variants that compensate for fitness costs of rifampicin
using the Illumina MiSeq BenchTop Genome Sequencer was
resistance mutations in rpoB. MDR strains with these compensatory
established and the new “Borstel Center for Genome Based
mutations exhibited a high competitive fitness in vitro and in vivo,
Molecular Epidemiology of Tuberculosis (BCGET)” was founded.
Using Nextera XT library preparation kits, protocols have been
Rhode Univ. of Central Florida, Orlando; P. Supply, Inst. Pasteur,
indicated above the gene diagrams of rpoA (a) and rpoC (b). Mutations identified by screening a global and a high-burden collection of
MDR strains are indicated by stars below the gene diagrams. Colors
in Germany; TBPANNET Consortium: 27 partners in Europe
and US; EUMEDNETvsTB: 7 partners in Europe and Africa;
indicate the respective strain lineage (blue, lineage 2; red, lineage 4;
PathoNgenTrace: 7 partners in Europe; Global Microbial
as determined by examining their relative clinical frequency across
brown, lineage 5; pink, lineage 1). Some of these mutations occurred
Identifier: More than 20 partners worldwide.
patient populations. These results suggest that the acquisition of
in multiple lineages or affect the same codon position (Comas et al.
developed that allow NGS analyses of clinical isolates from less
particular mutations over time in rpoA and rpoC in rifampicin-
Nature Genetics 2012 44, 106–110 (2012) doi:10.1038/ng.1038).
than 10 ng DNA, thus paving the way towards direct analysis of
resistant M. tuberculosis strains lead to the emergence of MDR
the transition of next generation sequencing to a highly efficient
BMBF project: Network “Pulmonary Tuberculosis – Host and
clinical samples. Currently, we are investigating several outbreaks
strains with high fitness. Our findings indicate that mathematical
technology for pathogen typing and diagnostics of three model
Pathogen Determinants of Resistance and Disease Progression
with NGS to define reliable parameters for the application of
models that aim at predicting the future of the global MDR TB
pathogens of worldwide importance MRSA, (Campylobacter spp.,
(TBornotTB)”; BMBF project: German Center for Infection Research
genome based molecular epidemiology in future studies.
epidemic should take into account the effects of compensatory
M. tuberculosis complex.) Within the German Center for Infection
(DZIF); EU-FP7 project (223681): Pan-European network for the
Molecular determinants and microevolution of resistant strains.
mutations as well as the time necessary for such mutations to
Research , FZB will lead the TTU tuberculosis with a focus of
study and clinical management of drug resistant tuberculosis;
The work on resistance mechanisms of clinical MTBC isolates has
emerge. Further studies are ongoing to investigate the importance
translational research in diagnostics, epidemiology, and clinics.
EU-FP7 project (245872): EUMEDNETvsTB Building a cooperative
been continued. By combining classical PCR-sequencing and
of compensatory mutations in different geographical settings.
new NGS based approaches, we have determined resistance
Grant application. Over the last two years we received two major
selected puBlications and Awards
upgrading tuberculosis research and control; EU-FP7 project
mechanisms for first line drugs (isoniazid, ethambutol), second line
grants: The EU-FP7-project Patho-Ngen-Trace is coordinated by
Roetzer A, Schuback S, Diel R, Gasau F, Ubben T, di Nauta A,
(278864): PathoNgenTrace: Next generation genome based high
drugs (prothionamide, linezolid), and new drugs in development
our group. Together with 7 partners in Europe, we will work on
Richter E, Rüsch-Gerdes S, Niemann S. Evaluation of Mycobacterium
resolution tracing of pathogens (Coordinator).
Grant support
strategy between Europe and Mediterranean countries for
Priority Research Area Infections • Molecular Mycobacteriology
PD Dr. Stefan Niemann
Clinical Infectious Diseases
Michael Block • Franziska Daduna • Cordula Ehlers • Emilia Häussinger • Dr. Christian Herzmann
clinical immunology
diagnosis
IGRA
MDR-TB
Dr. Gunar Günther •Christian Gutsfeld • Dr. Claudia Jafari • Dr. Barbara Kalsdorf • Ioana Olaru
Nelleke Smitsman
tuberculosis
prevention
MISSION
controls. CCR5 expression and RANTES concentration correlated
Dr. D.M. Cirillo (Milan, Italy), Dr. D. Falzon (Geneva, Switzerland);
To improve the prevention, diagnosis and treatment of tuberculosis and the integration of scientific
advances into clinical practice.
strongly with HIV-1 viral load in the BAL. In contrast, these altera-
Dr. N. Harada (Tokyo, Japan), Dr. E. Ibrahim (Bucharest, Roma-
tions were not associated with M. tuberculosis sensitisation in
nia), Dr. I. Katjitae (Windhoek, Namibia), Dr. C.C. Leung (Hong
vivo, nor did M. tuberculosis infection of BAL cells ex vivo change
Kong, China), Dr. F. v. Leth (Amsterdam, Netherlands), Prof. Dr.
most important findings
infection was assayed by an Interferon-γ Release Assay (IGRA)
RANTES expression. These data suggest ongoing HIV-1 replica-
D. Menzies (Montreal, Canada), Prof. Dr. H. Rieder (Zürich, Swit-
Defining the optimal treatment for patients with MDR-TB
in health care workers regularly exposed to tuberculosis pa-
tion predominantly drives local pulmonary CCR5+ T-cell activa-
zerland), Dr. A Skrahina (Minsk, Belarus), Prof. Dr. G. Sotgiu (Sas-
Treatment of multidrug resistant tuberculosis (MDR-TB) is lengthy,
tients and in household TB contacts in Germany. Immunopheno-
tion in HIV/latent M. tuberculosis co-infection.
sari, Italy), Prof. Dr. G. Walzl (Stellenbosch, South Africa), Prof.
toxic, expensive, and has generally poor outcomes. We under-
types of bronchoalveolar lavage mononuclear cells and periph-
took an individual patient data meta-analysis to assess the im-
eral blood mononuclear cells were analyzed by fluorescence
selected puBlications
(Hong Kong, China), Dr. J.-P. Zellweger (Berne, Switzerland); Col-
pact on outcomes of the type, number, and duration of drugs
activated cell sorting. All tuberculosis contacts with presumed
The Collaborative Group for Meta-Analysis of Individual
laborative Group for Meta-Analysis of Individual Patient Data
used to treat MDR-TB. Three recent systematic reviews were
latent infection (n=15) had increased (p<0.0001) frequencies
Patient Data in MDR-TB. Multidrug-resistant pulmonary tuber-
in MDR-TB; Tuberculosis Network European Trialsgroup (TBNET),
used to identify studies reporting treatment outcomes of micro-
of CD4+CD25+CD127- regulatory Treg cells (median 2.12 %, In-
culosis treatment regimens and patient outcomes: an individual
University of Namibia School of Medicine (Windhoek, Namibia);
biologically confirmed MDR-TB. Study authors were contacted to
terQuartile Range – IQR- 1.63-3.01) among bronchoalveolar
patient data meta-analysis of 9153 patients. PLoS Medicine
University of Medicine and Pharmacy and Institute for Ftizio-
solicit individual patient data including clinical characteristics,
lavage (BAL) mononuclear cells compared to contacts (n= 25)
2012; Aug;9(8):e1001300
pneumologie, Chisinau (Rep. Moldova); RESIST TB.
treatment given, and outcomes. Random effects multivariable
with negative IGRA test results (median 0.68 %, IQR 0.32-0.96)
logistic meta-regression was used to estimate adjusted odds of
In contrast, no differences were seen when peripheral blood
treatment success. Adequate treatment and outcome data were
mononuclear cells immunophenotypes of IGRA positive and
giu G, Lange C. Increased frequencies of pulmonary Treg cells
provided for 9,153 patients with MDR-TB from 32 observational
negative TB contacts were compared (IGRA+: median 9.6 %,
in latent M. tuberculosis infection. Eur Respir J 2012; 40: 1450-57
studies. Treatment success, compared to failure/relapse, was
IQR 5.9-10.1; IGRA-: median 7.7 %, IQR 4.6-11.3; p=0.47). Nota-
associated with use of: later generation quinolones, (adjusted
bly, 5 of the 25 contacts with negative blood IGRA showed a
Kalsdorf B, Skolimowska K, Scriba TJ, Rod Dawson, Dhe-
odds ratio [aOR]: 2.5 [95 % CI 1.1–6.0]), ofloxacin (aOR: 2.5 [1.6–
positive IGRA from BAL cells, possibly indicating a limited local
da K, Wood K, Hofmeister J, Hanekom WA, Lange C, and
3.9]), ethionamide or prothionamide (aOR: 1.7 [1.3–2.3]), use of
immune response. In Germany, presumed latent infection with
Wilkinson RJ. Relationship between chemokine receptor ex-
four or more likely effective drugs in the initial intensive phase
M. tuberculosis, as defined by the presence of a positive M.
pression, chemokine levels and HIV-1 replication in the lungs of
(aOR: 2.3 [1.3–3.9]), and three or more likely effective drugs
tuberculosis-specific IGRA response on cells from the peripheral
persons exposed to Mycobacterium tuberculosis. Eur J Immunol
in the continuation phase (aOR: 2.7 [1.7–4.1]). Similar results
blood, is characterized by an increased frequency of Treg cells in
2012 doi: 10.1002/eji.201242804
were seen for the association of treatment success compared
the bronchoalveolar lavage fluid.
to failure/relapse or death: later generation quinolones, (aOR:
Ongoing viral replication drives pulmonary T-cell activation
Lange C & Migliori G. European Respiratory Monograph #58
2.7 [1.7–4.3]), ofloxacin (aOR: 2.3 [1.3–3.8]), ethionamide or pro-
in HIV/latent M. tuberculosis co-infection
(2012); Tuberculosis
thionamide (aOR: 1.7 [1.4–2.1]), use of four or more likely effec-
Increased susceptibility to tuberculosis following HIV-1 serocon-
tive drugs in the initial intensive phase (aOR: 2.7 [1.9–3.9]), and
version contributes significantly to the tuberculosis epidemic in
internal and external collaborations
three or more likely effective drugs in the continuation phase
sub-Saharan Africa. Lung-specific mechanisms underlying the
Local: Prof. Dr. S. Ehlers, Dr. M. Ernst, Prof. Dr. C. Hoelscher, Prof.
(aOR: 4.5 [3.4–6.0]). In this individual patient data meta-analysis
interaction between HIV-1 and Mycobacterium tuberculosis in-
Dr. A. Katalinic, Prof. Dr. K-F. Klotz, PD Dr. S. Niemann, PD Dr.
of observational data, improved MDR-TB treatment success and
fection are incompletely understood. Here we address these
N. Reiling, Dr. S. Ruesch-Gerdes, Prof. Dr. U. Schaible, Prof. Dr.
survival were associated with use of certain fluoroquinolones,
questions by examining the effect of HIV-1 and latent M. tuber-
J. Westermann, Center for Infection and Inflammation Lübeck,
ethionamide, or prothionamide, and greater total number of ef-
culosis co-infection on the expression of viral-entry receptors and
Center for Infectious Diseases Borstel-Lübeck.
fective drugs. However, randomized trials are urgently needed
ligands in bronchoalveolar lavage (BAL) of HIV-1-infected and
National: C. Giehl (Saarbrücken), Prof. Dr. M. Hoelscher (Mu-
to optimize MDR-TB treatment.
-uninfected patients with and without latent M. tuberculosis infec-
nich), Prof. Dr. S.H.E. Kaufmann (Berlin), Prof. Dr. J. v. Lunzen
Pulmonary immunophenotype characterizes individuals
tion. Irrespective of HIV-1 status, T cells from BAL expressed high-
(Hamburg); Prof. Dr. M. Sester (Homburg), Prof. Dr. S. Stenger
Figure 2: Participants of the Borstel course on clinical tuberculosis, June
with latent M. tuberculosis infection
er levels of the beta-chemokine receptor (CCR)5 than peripheral
(Ulm), Dipl. Math. O. Vollrath (Kiel), PD Dr. D. Wagner (Freiburg);
14.-16. 2012
Regulation of pathogen specific immune responses following
blood T cells, in particular the CD8+ T cells of HIV-1-infected per-
DZIF; TB or not TB network.
airway contact to M. tuberculosis in humans and the role of
sons showed elevated CCR5 expression. The concentrations of
International: Dr. E. Bond (Stockholm, Sweden), Dr. J. Camine-
Grant support
regulatory T (Treg) cells in immune control of latent infection with
the CCR5 ligands RANTES and MIP-1β were elevated in the BAL
ro, Las Palmas de Gran Canaria, Spain), Prof. Dr. P.-J. Cardona
BMBF, DAAD, DZIF, EDCTP, EU-FP7, ExcellenceCluster i@i, S.-H.
M. tuberculosis are incompletely understood. Presumed latent
of HIV-1-infected persons compared with that in HIV-1-uninfected
(Barcelona, Spain), Dr. V. Crudu (Chisinau, Rep. Moldova), Prof.
TB Society, TBNET; AID, Alere, Siemens
Dr. R. Wilkinson (Cape Town, South Africa), Prof. Dr. W.W. Yew
Herzmann C, Ernst M, Ehlers S, Stenger S, Maertzdorf J, Sot-
Figure 1: The optimal duration of therapy in MDR-TB appears to be >
24 months on average with currently available therapies (PLoS Medicine 2012; Aug;9(8):e1001300)
53
Priority Research Area Infections • Clinical Infectious Diseases
Prof. Dr. Christoph Lange, FIDSA
XDR-TB
Dr. Jan Heyckendorf • Jessica Hofmeister • Christiane Gerlach • Dr. Ulf Greinert • Susanne GroSSmann
Experimental Pneumology
allergic bronchial asthma
nanotoxicology
Dr. Christina Vock • Dr. Sandra Schlick • Sina Webering • Franziska Beyersdorf • Juliane Artelt • Kerstin
Viertmann • Carina Krützmann
alveolar regeneration
design-based stereology
airway remodeling
MISSION
ronments. We investigate whether carbon black nanoparticles
Seifart C, Muyal JP, Plagens A, Yildirim AÖ, Kohse K, Grau
To contribute to a better understanding of the role of airway as well as alveolar epithelium in the
pathogenesis of chronic inflammatory lung diseases such as allergic bronchial asthma, COPD/emphysema, and inflammatory processes related to nanoparticle exposure.
(CBNP), which are present in industrially produced soot used
V, Sandu S, Reinke C, Tschernig T, Vogelmeier C, Fehren-
for reinforcement of elastomers but also for paints, toner and
bach H. ATRA results in irregular repair of septa and fails to in-
batteries, may have cytotoxic and pro-inflammatory effects in
hibit proinflammatory macrophages. EUROPEAN RESPIRATORY
the airways of mice and whether differences are seen in the
JOURNAL 2011 Aug;38(2):425-39.
most important findings
goblet cell metaplasia. Effects of IL-13 are mediated by the IL13
response of proximal versus distal airways. Further, we aim at
Allergic bronchial asthma is one of the most common chronic
receptor (IL-13R) composed of the IL-4Rα and IL-13Rα1 subunits.
investigating whether surface modifications on CBNP can lead
internal and external collaborations
diseases in the world and developed to a major health prob-
We therefore investigated whether differences in the IL-13/IL-13R
to a modification of their biological effects. The impact of CBNP
Close collaborations within the Research Center Borstel are es-
lem of the western world. According to the WHO-funded Global
signalling cascade are responsible for airway-region specific
on different airway regions was assessed by microdissection
tablished with the Divisions of Mouse Models of Asthma, Inver-
Initiative for Asthma Report on the Global Burden of Asthma as
mucus production in mice sensitized subsequently challenged
of proximal and distal airways from mouse lungs and ex vivo
tebrate Models, Mucosa Immunology and Diagnostics, Immuno-
many as 300 Million people suffer from asthma. The tremendous
with OVA for one (acute) or twelve weeks (chronic). Lungs were
stimulation for 0.5–48 hours. Cytotoxicity was measured by LDH
biophysics and Clinical and Experimental Pathology.
impact of asthma on health care systems is demonstrated by
inflated with RNAlater® to microdissect proximal and distal air-
release and cytokine mRNA expression by qRT-PCR. Our study
Several research projects are pursued in cooperation with
the annual direct medical and indirect economic costs (ie loss
ways for qRT-PCR. Laser-assisted microdissection (LCM) of air-
could not show any cytotoxic effects on microdissected airways
groups at the University of Lübeck: Peter König, Institute of Anat-
of work days, lost productivity, premature retirement), which ac-
way epithelial cells was performed on cryofixed lungs. qRT-PCR
after incubation with Printex 90 or acetylene carbon black which
omy; Gereon Hüttman, Institute for Biomedical Optics; Rudolf
cumulate to approximately Euro 17.7 Billion for Europe. In Ger-
analyses revealed significantly lower abundance of IL-13Rα1 in
contains PAHs on its surface (Figure 2).
Manz, Institute for Systemic Inflammation Research; University
many approx. 368.000 work days per year are reclusively lost due
distal airways, whereas mRNA levels of IL-4Rα were similar in
of Kiel: Stefan Schütze, Institute of Immunology; Martin Krause,
to asthma ranging almost on the same level like diabetes and
proximal and distal airways. Investigations of major transcrip-
Department of Pediatrics University Medical Center Schleswig-
chronic-ischemic heart diseases.
tion factors down-stream the IL-13/IL-13R signalling, displayed
Holstein.
One of the hallmarks of allergic bronchial asthma is the remod-
a significant airway-region specific gene expression pattern: i)
Close external collaboration has been established with the
eling of the airway wall which comprises epithelial goblet cell
expression levels of Spdef and FoxA3, both known to promote
partners of the BMBF funded joint project “CarbonBlack”: Hen-
metaplasia, subepithelial fibrosis, and smooth muscle hyperpla-
goblet cell metaplasia and mucus production, were barely ex-
ning Bockhorn (Karlsruhe Institute of Technology), Bernd Müller
sia. We were able to demonstrate that airway remodeling is
pressed in distal airways but significantly up-regulated in proxi-
(Philipps-University of Marburg), Tanja Hansen (Fraunhofer Insti-
markedly reduced in distal airways as compared to proximal
mal airways of OVA-treated mice; ii) repressors of goblet cell
tute of Toxicology and Experimental Medicine, Hannover) and
airways in mice (Figure 1) despite comparable degrees of in-
metaplasia and mucus production, namely FoxA2 and Nkx2-1,
Peter König (University of Lübeck) as well as with various collabo-
filtration and expression of IL-13, which is the main trigger for
were both higher expressed in distal airways of PBS mice and
repressed in proximal and distal airways of OVA-treated mice.
rators at partner-sites of the German Center for Lung Research:
Figure 2: Incubation of microdissected proximal (blue bars) and distal
(red bars) airways with increasing concentrations of quartz particles
Another hallmark of allergic bronchial asthma is a persistent
inflammation of the airway wall. Recent studies identified T
helper 17 cells as important players in the progression of asth-
(DG12) or the carbon black nanoparticles Printex90 and acetylene
soot with surface PAHs (Ac-Ruß mit PAKs) does not induce cytotoxicity
as measured by LDH assay. Incubation with Tween was used as a po-
Functional Anatomy, Hanover Medical School), Marcus Mall (Department of Translational Pulmonology, Heidelberg).
Grant support
tion of the pro-inflammatory cytokines TNF-α, IL-1ß, IL-22 and
selected publications
BMBF: German Center of Lung Research, co-coordinator of Dis-
IL-17A, TH17 cells appear to act as general promoters of chronic
Wegmann M, Lunding L, Orinska Z, Wong DM, Manz RA,
ease Area Asthma/Allergy (H.F.);
inflammatory responses. TH17 cells are not only characterized
Fehrenbach H. Long-Term Bortezomib Treatment Reduces Aller-
co-cordinator of Platform Imaging (H.F.)
by the expression of specific cytokines, but also by the exclu-
gen-Specific IgE but Fails to Ameliorate Chronic Asthma in Mice.
BMBF: Consortium “Carbon Black” (FKZ 03X0093A), coordina-
sive expression of the t-splice variant of the transcription factor
INTERNATIONAL ARCHIVES OF ALLERGY AND IMMUNOLOGY
tor (H.F.)
RORγ, which is a homologue to RORγ. Therefore, in another proj-
2012;158(1):43-53.7
DFG: Cluster of Excellence “Inflammation at Interfaces”
Figure 1: Regional differences in mucus production in a mouse model
ect, which is conducted in co-operation with the research group
of allergic bronchial asthma. Whereas in proximal airways, which are
“Mouse Models of Asthma”, we are investigating the suitability
Kılıç A, Sonar SS, Yildirim AO, Fehrenbach H, Nockher WA,
characterized by significant expression of IL-13Ra1, marked production
of RORγt as a therapeutic target in severe asthma.
Renz H. Nerve growth factor induces type III collagen produc-
down-stream transcription factors Spdef and FoxA3, distal airways
Medicine, Hannover), Matthias Ochs (Institute for Applied and
sitive control and culture medium without particles as negative control.
ma towards a severe phenotype. Characterized by the produc-
of mucus is seen to be associated with increased expression of the
Armin Braun (Fraunhofer Institute of Toxicology and Experimental
(EXC306-DW2, EXC306-STI, EXC306CL10)
tion in chronic allergic airway inflammation. JOURNAL OF AL-
show minimal expression of all these components of the IL-13 signaling
Exposure to airborne ultrafine or nanoparticles is typically
LERGY AND CLINICAL IMMUNOLOGY 2011 Nov;128(5):1058-
cascade with no mucus production despite equivalent levels of IL-13.
related to urban environments or specific occupational envi-
U510.
55
Priority Research Area Asthma & Allergy • Experimental Pneumology
Prof. Dr. Heinz Fehrenbach
Molecular Immunology
Influenza virus
induced airway
inflammation
Andreas Baier • Weijia Cheng • Dr. Niko Föger • Martina Hein • Manuel Hein • Azhar Kamal • Lekha Nath
Patricia Prilla •Jin-Bo Yu
TNFα signaling
Kyeong-Hee Lee, PhD
MISSION
Nguyen XH, Lang PA, Lang KS, Fattakhova G, Adam D, Föger
Our overall research goal is to explore novel regulatory themes in inflammatory signal transduction. In a multidisciplinary approach we aim at identifying molecular and cellular key mechanisms underlying the development of immune disorders, such as inflammatory disease, as well as determining
perspectives for clinical interventions
N, Kamal A, Prilla P, Mathieu S, Wagner C, Mak T, Chan
most important findings
Our group has successfully generated the first TOSO-deficient
Lang K, Lang P, Meryk A, Pandyra A, Boucher LM, Pozdeev
The role of anti-apoptotic molecule TOSO in controlling
mice. TOSO-/- T cells show defects in TNFα-mediated functions,
V, Tusche M, Göthert J, Haight J, Wakeham A, You-Ten A,
adaptive immune responses to viral infection and the
such as increased sensitivity to TNFα-induced apoptosis and
McIlwain D, Merchers K, Khairnar V, Recher M, Nolan G,
development of virus induced airway inflammation
reduced TNFα-mediated survival signaling. An in vitro analysis
Hitoshi Y, Funkner P, Navarini A, Verschoor A, Shaabani N,
The regulation of lymphocyte apoptosis is critical to immune cell
of B cell proliferation and survival has revealed dramatically
Honke N, Penn L, Ohashi P, Häussinger D, Lee KH* and
homeostasis and normal functioning of the immune system. Even
reduced antigen receptor-mediated cell proliferation, which
Mak TW* (*shared senior authorship). Involvement of Toso in
a small imbalance in the regulation of lymphocyte survival and
is associated with a significantly higher degree of cell death
activation of monocytes, macrophages and granulocytes. Proc
apoptosis can have severe pathological consequences, such as
in B cell cultures from TOSO-/- mice compared to WT cultures
Natl Acad Sci U S A. 2013 Jan 28. [Epub ahead of print].
inflammatory disease or autoimmunity. The recently described
(Figure A, B). Consistent with an anti-apoptotic function of TOSO,
surface molecule TOSO (Faim3), which is highly expressed in
increased cell death was also observed in cultures of activated
internal and external collaborations
B cells and activated T cells, has been identified as an anti-
TOSO-/- T cells (Figure. C).
Dr. Tak Mak, Campbell Family Institute for Breast Cancer
and non-apoptotic death receptor signaling by facilitating RIP1
ubiquitination. BLOOD, 2011 Jul 21;118(3):598-608.
Research, Toronto, Canada
TOSO exhibits its anti-apoptotic effects by a novel regulatory
Given the anti-apoptotic function of TOSO and the critical role
Prof. Dr. Dieter Adam, University of Kiel, Germany
mechanism involving the adaptor kinase RIP1 and an ubiquitin
of Fas and TNFα for adaptive immune responses, we have
Dr. Karl Lang, Humboldt Reserch Group, University of Düsseldorf,
pathway. Ultimately, TOSO exhibits its anti-apoptotic activity
investigated the overall role of TOSO in controlling the survival of
Germany
by facilitating death receptor-induced pro-survival signals. Our
effector T and B cells during viral infection. Our data indicate that
Dr. Andrew C. Chan, Vice President Research Immunology,
data on the mechanistic analysis of the anti-apoptotic function of
TOSO contributes to T and B cell expansion and maintenance
Genentech Inc., USA
TOSO and the critical role of TOSO in death receptor signaling
of germinal center (GC) B cells during the expansion phase
Prof. David Hildemann: Cincinnati Childrens‘s Hospital Medical
resulted in two Blood paper publications (Nguyen et al., Blood,
upon infection with lymphocytic choriomeningitis virus (LCMV)
Center, USA
2011; Nguyen et al., Blood, 2012).
Armstrong as well as clone 13 (unpublished data). Preliminary
studies also indicate that TOSO-/- mice are largely resistant to
Grant support
influenza A induced airway inflammation. Upon influenza A
Marie Curie International Reintegration Grant: FP7-PEOPLE-
infection TOSO-/- mice exhibit impaired immune cell recruitment
2007-4-3-IRG (2007-2011)
to the lung and show substantially reduced production of pro-
Fritz Thyssen Research Foundation: Az. 10.08.2.155 (2009-2011)
inflammatory cytokines (unpublished data). Moreover, our lab
German Research Foundation (DFG): LE 1254/2-1 (2008-2011)
has successfully developed a set of TOSO-specific monoclonal
German Research Foundation (DFG): LE 1254/2-2 (2011-2014)
antibodies that specifically block the anti-apoptotic function of
German Research Foundation (DFG): LE 1254/3-1 (2012-2015)
TOSO and inhibit TNFα-mediated survival signaling pathways.
Cluster of Excellence in Inflammation Research (2009-2012)
Due to these unique features, these TOSO specific blocking
antibodies have great potential to lead to the development
Figure 1: Defective cell proliferation and increased cell death in
of novel clinical applications for the treatment of inflammatory
cultures of lymphocytes from TOSO mice. (A) CFSE dilution analysis
disorders.
-/-
of B cells from WT and TOSO-/- mice after stimulation with anti-IgM. (B) B
cells from WT and TOSO-/- mice were stimulated with anti-IgM or IL-4 and
cell death was assessed after 48h by staining with the cell viability stain
7-AAD. (C) T cells from WT and TOSO-/- mice were activated by anti-CD3/
selected puBlications
Nguyen XH, Fattakhova G, Lang PA, Lang KS, Adam D,
anti-CD28 stimulation and cell death was monitored over time by staining
Föger N, Lee KH. Antiapoptotic function of Toso (Faim3) in
with 7-AAD
death receptor signaling. BLOOD, 2012 Feb 16; 119(7):1790-1 .
57
Priority Research Area Asthma & Allergy • Molecular Immunology
apoptotic regulator of death receptor (Fas and TNFRI) signaling.
AC, Lee KH. Toso regulates the balance between apoptotic
Biochemical Immunology
Mast cell-neutrophil interaction
asthma
Christine Engellenner • Cindy Hass • Diana Heinrich • Gabriele Huss • Dr. Brigitte Kasper • Anika Kasprick
Junie Tchudjin Magatsin • Reza Akbarzadeh Najar • Carola Schneider • Xinhua Yu , PhD
autoimmunity
tissue damage
MISSION
temic passive model of EBA. After repeated administration of
ase reporters. These molecules contain a lipid anchor for mem-
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 dermatoses.
pathogenic anti-murine collagen 7-IgG, mice developed the typi-
brane integration and a protease-specific cleavage sequence
cal phenotype of EBA already within four days after first antibody
conjugated to a FRET-pair of fluorochromes (Fig.2). Cleavage by
injection. However, unlike our hypothesis, none of the MC--strains
corresponding proteases results in the separation of both fluoro-
were protected from disease. Unexpectedly, EBA was dramati-
chromes leading to the loss of FRET, which can be analyzed by
cally increased in C57BL/6J-kitW-sh/kitW-sh while the disease states
confocal and multiphoton microscopy. We will use this technique
to visualize neutrophil protease (neutrophil elastase and MMP9)
most important findings
sors results in neutropenia due to death during differentiation. If
of WBB6F1/J-kit /kit
Allergic asthma appears as a chronic eosinophilic inflamma-
successful, this model will be used in acute and chronic models
comparable. To clarify these opposing results possibly caused
tion of the airways which was, until recently, seen as directly
of asthma as well as in our models of autoimmune dermatoses.
by unknown side effects of the KIT mutations, the role of MC
causative for the asthma pathology in terms of airway hyperreac-
The question whether mast cells could be involved in the recruit-
was further investigated in the transgenic Mcpt5-Cre iDTR strain.
selected puBlications
tivity and remodelling. However, the role of eosinophils appears
ment of neutrophils and eosinophils into the allergic lung was
Thereby, no significant difference between MC-deficient and
Kasper B, Petersen F. Molecular pathways of platelet factor 4/
to be less clear today. Eosinophilia is rarely observed in severe
addressed first by a series of experiments in vitro using human
MC-sufficient groups could be observed confirming the findings
CXCL4 signaling. European Journal Of Cell Biology. 2011; 90:521-
asthma and, according to most recent studies, nearly 50 % of the
primary lung mast cells. We could show that mast cells secrete
of the WBB6F1 KitW/KitW-v mice. These results obtained in the sys-
526.
mild to moderate asthmatics are characterized by normal levels
immediately after stimulation high levels of LTB4 and PGD2
temic passive model of EBA could be confirmed in a recently
of eosinophils in their sputum. Since asthma and several other
which induces chemotaxis, transmigration, adhesion, and ROS-
established local model of the disease, which was used previ-
Schwartzkopff F, Petersen F, Grimm TA, Brandt E. CXC chemo-
chronic airway inflammations are associated with an increase of
production in neutrophils and eosinophils. However, after 18 to
ously to analyze the role of MC in experimental BP. In summary,
kine ligand 4 (CXCL4) down-regulates CC chemokine receptor
neutrophils, we investigated the pathophysiological role of these
24h of stimulation a second neutrophil- and eosinophil-activating
our results indicate that, in contrast to BP, in EBA MC do not
expression on human monocytes. Innate Immunity. 2012; 18:124-
cells in two different mouse models of acute allergic asthma.
mediator became apparent in the supernatants. Due to its bio-
contribute to the pathogenesis of the disease. Although closely
139.
Depletion of neutrophils by antibodies was accompanied by a
chemical properties, we could characterize this mediator as a
related, treatment of both diseases may require principally dif-
reduced eosinophilia in the BAL and less inflammatory cellular
protein with a molecular weight of 30-50 kD which consists of at
ferent therapeutical strategies.
infiltrates in the lung tissue. Furthermore, goblet cell hyperplasia
least two different subunits and does not belong to the family of
and airway hyperreactivity were significantly decreased in the
chemokines. After having identified this molecule we will evalu-
Neutrophils proteases are also known to promote tissue dam-
depends on the localization of the affected tissue. Genes & Im-
latter animals (Fig.1).
ate both mediators in sputum and BAL samples derived from
age and remodeling in chronic inflammatory processes like
munity. 2012 Dec;13(8):641-52.
asthma patients as well as in the lungs of mice with experimen-
asthma, COPD, or autoimmunity of the skin. Until now, analysis
tal allergic lung inflammation.
of neutrophil activation in terms of protease release is limited to
internal and external collaborations
the determination of the enzymes or the corresponding enzyme
Mast cell-neutrophil interaction in asthma - E. Brandt, O. Holst, B.
Epidermolysis bullosa acquisita (EBA) and Bullous pemphi-
activity in solution. To enable a site-specific analysis of neutro-
Lindner, A. Petersen, Research Center Borstel; H. Renz, H. Garn,
goid (BP) are acquired chronic subepidermal blistering disease
phil degranulation on single-cell level in diseased tissue, organs,
L. Marsh, University of Marburg; J. Köhl, ISEF, P. König, Institute
of skin and mucous membranes characterized by circulating and
or living animals we are currently establishing a method using
of Anatomy, University of Lübeck. S100A8/A9 proteins in asthma
tissue-bound autoantibodies against type VII (EBA) or type XVII
lipidated FRET (fluorescence resonance energy transfer) prote-
and autoimmunity - T. Vogl, University of Münster. FRET protease
Figure 1: Neutrophil-depletion by Gr-1 antibodies reduces allergic
airway inflammation and goblet cell hyperplasia.
W
W-v
mice and their wild type controls were
activity in experimental models of autoimmunity and asthma.
Zheng J, Ibrahim S, Petersen F, Yu X. Meta-analysis reveals an
association of PTPN22 C1858T with autoimmune diseases, which
collagen (BP). In our studies we analyze the contribution of mast
reporters - T. Scholzen, Research Center Borstel; C. Schultz, EMBL
cells and neutrophils to the pathogenesis of the diseases.
Heidelberg. Autoimmune dermatoses - D. Zillikens, R. Ludwig,
The role of mast cells (MC) in autoimmune diseases has become
Institute of Dermatology, University of Lübeck.
apparent only recently. Using MC-deficient mouse strains (MC-strains), several studies could allocate a disease-promoting func-
Grant support
Surprisingly, the reduced asthma pathology was not associated
tion of MC in experimental models of multiple sclerosis, rheuma-
DFG SFB/TR 22 “Allergische Immunantworten der Lunge“, Teil-
by changes in TH2 cytokine expression, indicating that TH2 cells
toid arthritis, and bullous pemphigoid (BP). However, most recent
projekt A11.
may not be involved in this process. Since depletion of cells
results suggest that the outcome of such experiments strongly
DFG Cluster of Excellence „Inflammation at Interfaces”; Research
by antibodies is always limited by their specificity, we are cur-
depends on the individual MC--strains used. In order to evaluate
Area H: IRN Autoimmunity to Type VII Collagen.
rently establishing a animal model of neutropenia using LysM-
MC as potential therapeutical targets we have analyzed their
DFG GRK 1727 „Modulation von Autoimmunität“ Teilprojekt 12
Cre Mcl-1fl/fl transgenic mice. Mcl-1 represents an anti-apoptotic
role in the pathogenesis of EBA. To determine their role in EBA in
BMBF Deutsches Zentrum für Lungenforschung, ARCN
factor which is involved in the terminal differentiation of many
general, three mouse strains (WBB6F1 KitW/KitW-v, C57BL/6J kitW-sh/
SPP AI „Mechanismen und innovative Modulation von organspe-
cell types. Lineage-specific deletion of Mcl-1 in myeloid precur-
kit
, Mcpt5-Cre iDTR) were investigated comparatively in a sys-
W-sh
Figure 2: The elastase-specific lipidated FRET reporter NEmo2.
zifischer Autoimmunität“, Teilprojekt B4.
59
Priority Research Area Asthma & Allergy • Biochemical Immunology
Prof. Dr. Frank Petersen
Invertebrate Models
Drosophila
Carina Krützmann
Stress
neuroendocrine system
innate immune response
asthma model
MISSION
be investigated that might have a systemic effect on the innate
Grant support
to elucidate the interaction between stress, innate immune system, and its systemic effect on the respiratory defense response in order to better understand to which extend stress contributes to the
development, aggravation, and pathogenesis of chronic inflammatory diseases in humans such as
bronchial asthma and chronic obstructive pulmonary disease (COPD).
immune response in general and that of the airways in particu-
German Research Foundation (DFG): WA 2972 2/2
lar. Subsequently, neuronal and endocrine transmitters will be
deciphered that transfer neuronal and endocrine signals to immune-competent cells. Relating to this issue, different transgenic Drosophila models should be generated to map the most
most important findings
naling pathway in airway epithelial cells and mainly involved in
important physiological stress axis (e. g. neuronal pathways or
Neuronal and endocrine transmitters modulate a variety of
the expression of antimicrobial peptide genes. By using the so-
hormonal circuits) that produce the newly identified mediators
physiological functions including the immune system. Stress
called targeted gene expression system Gal4/UAS by Andrea
and secret them through innervation after stress exposure. A
has been identified as a central factor influencing these highly
Brand and Norbert Perrimon we could show additionally that
more detailed analysis of the neuroendocrine-immune crosstalk
complex interrelationships, which may influence in long-term
strong ectopic activation of this signaling pathway by overex-
will be supplemented by mechanistic analysis at the molecular
the initiation, progression, and aggravation of chronic inflamm-
pression of the pattern recognition receptor PGRP-LC or PGRP-
and cellular level of immune competent-cells in order to figure
atory diseases in humans such as COPD and bronchial asthma.
LE leads beside AMP expression to phenotypic alterations of
out which second messenger molecules (e. g. Ca2+ or cAMP)
In humans, extreme or repeated stress markedly affects these
the entire airway epithelium, as they could also partly be ob-
or immune signaling pathways (NF-κB, JNK) trigger the immune
immune inflammatory disorders by activating the hypothala-
served in the airways in response to strong bacterial infections.
alterations in the corresponding immune cells.
mic-pituitary-adrenal axis as well as the sympathetic-adrenal-
Phenotypic characteristics of ectopic NF-κB activation are me-
medullary system. The latter in particular is responsible for the
taplastic (thickening of airway epithelial cells) and hyperplastic
selected puBlications
synthesis and secretion of catecholamines (adrenaline, norepi-
(epithelial cell proliferation) alterations of the airway epithelium
Roeder T, Isermann K, Kallsen K, Uliczka K, Wagner C.
Figure 1: Structural alterations of the whole airway epithelium can be
nephrine) upon excitability, which in turn orchestrates the pro-
which also occur in the entire airways by overexpression of the
(2011) A Drosophila asthma model – what the fly tells us about
induced by ectopic activation of the NF-κB signaling pathway IMD.
duction and secretion of proinflammatory mediators through
transcription factor dFoxO. Downregulation of foxO via RNAi
inflammatory diseases of the lung. Adv Exp Med Biol. 710:37-47
immune-competent cells of the innate immune system. Since the
leads generally to the reduction of the epithelial thickness in
neuroendocrine as well as the immune system of vertebrates
an NF-κB active background supporting strongly the conclusi-
Roeder T, Isermann K, Wagner C, Warmbold C. (2011) Fruit
are very complex biological systems, the interactions of the
on that foxO is a downstream target of NF-κB. Interestingly, a
flies as a model in biomedical research – a Drosophila asthma
neuroendocrine, the respiratory and innate immune systems
comparable phenotype of airway epithelial cells is seen in the
model. Insect Biotechnology. 2:15-27
are very difficult to elucidate. In this context, the apparent sim-
pathological state of chronic inflammatory lung disease such
ple fruit or vinegar fly Drosophila melanogaster turns out as an
as bronchial asthma. Unfortunately, the molecular and cellular
Wagner C, Isermann K, Roeder T. (2009) Infection induces a
informative model since analogue and homologous structures
mechanisms that lead to these characteristic phenotypic altera-
survival program and local remodelling in the airway epitheli-
to the vertebrate neuroendocrine, respiratory as well as inna-
tions in humans are still not sufficiently understood. In this con-
um of the fly. FASEB J. 23:2045-54
te immune system exist and its genome largely lacks genetic
text Drosophila is a promising model to rapidly and specifically
redundancy.
decipher how these phenotypic alterations of airway epithelial
Wagner C, Isermann K, Fehrenbach H, Roeder T. (2008) Mo-
Microscopic (A,B) and electron microscopic (C,D) analyses of primary
airways from control animals (A,C) and those where the NF-κB signaling pathway IMD was ectopically activated by overexpression of the
pattern recognition receptor PGRP-LE (B,D).
cells might develop.
lecular architecture of the fruit fly‘s airway epithelial immune
Drosophila in asthma research: Since a few years it is well
Due to these findings and the fact the stress contributes to
system. BMC Genomics. 9: 446
known that in humans the airway epithelium plays a key role in
the development, exacerbation, and pathogenesis of bronchi-
the development, persistence, and pathogenesis of bronchial
al asthma in humans, we broadened our research field and
internal and external collaborations
asthma. In this context Drosophila is an ideal model organism
concentrate right now on the interaction between stress, innate
at the Research Center Borstel we cooperate with die Divisions
express dFoxO ectopically (B). Quantification of epithelial thickness in
not only in terms of its simplicity or easy genetic manipulation,
immune system, and its systemic effect on the innate immune
of Innate Immunity (PD Dr. Holger Heine), Experimental Pneu-
larva with different transgenic backgrounds (C).
but also due to the fact that the airway system of Drosophila
response of the airways.
mology (Prof. Dr. Heinz Fehrenbach), Asthma Mouse Models
consists primarily of airway epithelial cells. Therefore, Droso-
61
(Dr. Michael Wegmann), Microbial Interface Biology (PD. Dr.
phila is used as an ideal model for the investigation of the
Aims 2013/2014: In the following two years we would like to
Norbert Reiling)
epithelial immunity exclusively in-vivo.
investigate the interaction between stress and innate immune
External cooperations has been established with Prof. Dr. Tho-
Using this model we could demonstrate very strikingly, that the
system in more detail. First of all, we would like to identify stress
mas Roeder, Zoological Institute, CAU Kiel and Prof. Dr. Ulrich
NF-κB signaling pathway IMD (Immune Deficiency), a homo-
factors with an immune modulatory effect. In this context basic
Theopold, Department of Molecular Biology and Functional
logue of the TNF-α signaling pathway, is the central NF-κB sig-
stress factors such as starvation, heat, and cold stress should
Genomics, Stockholm University, Sweden
Figure 2: FoxO is mainly responsible for the remodeling of the airway
epithelium. Structural analyses of terminal airways by electron microscopy (A,B) that originate either from control animals (A) or animals that
Priority Research Area Asthma & Allergy • Invertebrate Models
Dr. Christina Wagner
Immune Cell-Analytics
immune cell regulation
Franziska Daduna
allergy
IFN-γ Elispot
mycobacterial infection
MISSION
Herzmann C, Lange C, Schaberg T, Ernst M, Ehlers S; TB or
We examined phenotypic and functional changes in human cells of the innate immune system (monocytes, basophils) and in cells of the adaptive immune system (memory T-lymphocytes) after in vitro
exposure to antigens or allergens. We aim to determine early signs of disturbed immunoregulation in
healthy individuals and in patients with infectious and / or inflammatory lung diseases as well as in
allergy patients.
Not TB Consortium.
Immunological evidence of incipient pulmonary tuberculosis. J
Infect Dis. 2012 Nov 15;206(10):1630-1; author reply 1631-2. doi:
10.1093/infdis/jis557. Epub 2012 Sep 10
Heinbockel L, Sánchez-Gómez S, Martinez de Tejada G,
most important findings
Dömming S, Brandenburg J, Kaconis Y, Hornef M, Dupont
Increased frequencies of pulmonary regulatory T-cells in latent
A, Marwitz S, Goldmann T, Ernst M, Gutsmann T, Schürholz
Mycobacterium tuberculosis infection. (in collaboration with
T, Brandenburg K. Preclinical Investigations Reveal the Broad
the Division of Clinical Infectious Diseases [C. Lange] and the
Spectrum Neutralizing Activity of Peptide Pep19-2.5 on Bacterial
Center for Clinical Studies at the Research Center Borstel [C.
Pathogenicity Factors. Antimicrob Agents Chemother. 2013 Jan
Herzmann])
14. [Epub ahead of print]
Background: Regulation of specific immune responses
internal and external collaborations
following exposure to Mycobacterium
Mycobacterial infections related projects are performed in close
tuberculosis in humans and the role of regulatory T (Treg) cells
internal collaboration with the Division of Clinical Infectious
in the immune control of latent infection with M. tuberculosis
Diseases (C.Lange), the Clinical Study Center (C.Herzmann) and
are incompletely understood.
the Outpatient Clinic (U.Greinert) and external collaborations
Methods: Latent infection was assayed by an interferon-
with members of the TBNET.
gamma release assay (IGRA) in peripheral blood mononuclear
Member of GOLDnet ( German Network for Diffus Parenchymal
cells (PBMC) of healthcare workers regularly exposed to
Lung Disease) with a focus on immunodiagnostics of berylliosis
tuberculosis (TB) patients and in household TB contacts in
(collaboration with GOLDnet members K.Gaede [Research
Germany.
Immunophenotypes
of
bronchoalveolar
lavage
(BAL)
Figure 1: Frequency of Treg cells in BAL-lymphocytes stratified by
PBMC IGRA status
Center Borstel] and G.Zissel [University of Freiburg])
Allergy related projects are performed in close collaboration
IGRA was performed in PBMCs. Black dots: paired PBMC and BAL
with the Division of Clinical and Molecular Allergology (U.Jappe,
cytometry in a FACS Calibur cytometer.
samples available. White dots: additional unpaired BAL and PBMC
A.Petersen)
Results: All TB contacts with latent infection (n=15) had
samples. Bars: median and interquartile range
The project C6 (Sleep supports the adaptive immune response to
mononuclear cells and PBMCs were analysed by flow
increased (p <0.0001) frequencies of CD4+CD25+ CD127- Treg
vaccination) of the Collaborative Research Center 654 Plasticity
cells (median 2.12 %, interquartile range (IQR) 1.63–3.01 %)
selected puBlications
and Sleep is jointly headed by T. Lange (UKSH Campus Lübeck)
among BAL mononuclear cells compared with contacts (n=25)
Jafari C, Kessler P, Sotgiu G, Ernst M, Lange C. Impact of
and M.Ernst (Division of Immune Cell-Analytics, Borstel)
with negative IGRA results (median 0.68 %, IQR 0.32–0.96 %).
a Mycobacterium tuberculosis-specific interferon-γ release
No differences were seen when PBMC immunophenotypes of
assay in bronchoalveolar lavage fluid for a rapid diagnosis
Grant support
IGRA+ and IGRA- TB contacts were compared (IGRA+ median
of tuberculosis. J Intern Med. 2011 Sep;270(3):254-62. doi:
Collaborative Research Center 654 Plasticity and Sleep
9.6%, IQR 5.9–10.1 %; IGRA- median 7.7 %, IQR 4.6– 11.3 %;
10.1111/j.1365-2796.2011.02378.x. Epub 2011 Apr 11. Erratum in: J
Project C6 (together with T.Lange, UKSH Campus Lübeck) Sleep
p=0.47). Five out of 25 contacts with negative blood IGRAs
Intern Med. 2011 Nov;270(5):500.
supports the adaptive immune response to vaccination
showed a positive IGRA from BAL cells, possibly indicating a
limited local immune response.
Herzmann C, Ernst M, Ehlers S, Stenger S, Maertzdorf J, Sotgiu
Conclusion: In Germany, latent infection with M. tuberculosis,
G, Lange C. Increased frequencies of pulmonary regulatory
as defined by a positive M. tuberculosis-specific IGRA
T-cells in latent Mycobacterium tuberculosis infection. Eur Respir
response in cells from the peripheral blood, is characterised
J. 2012 Dec;40(6):1450-7. doi: 10.1183/09031936.00214611. Epub
by an increased frequency of Treg cells in the BAL.
2012 Mar 22.
63
Priority Research Area Asthma & Allergy • Immune Cell-Analytics
Dr. Martin Ernst
Innate Immunity
Innate immunity
Suhad Al-Badri • Katrin Böhnstedt • Dr. Erwin Duitman • Ina Goroncy • Kimberley Kallsen • Dr. Thomas
Scholzen • Dr. Karina Stein • Anna Störmer • Dr. Karin Uliczka • Prof. Artur J. Ulmer
allergy
Drosophila
MISSION
The impact of environmental stress especially on barrier epi-
involves FoxO in an inhibiting function. With regard to our recent
The mission of the division of Innate Immunity is the investigation, analysis and characterization of
activation mechanisms of the innate immunes system through microbes, microbial structures and allergens and the interplay and intercommunication with the adaptive immune system. In particular, we
are interested in the consequences of this activation for lung diseases such as allergies and asthma.
thelia is a more and more approved cause of various epithe-
results, FoxO seems to be a key protein in cellular response
lial diseases such as chronic inflammation, COPD or Asthma
against stress and a link between stress input and immune reac-
bronchiale. Elucidating the reactions environmental stressors
tion. (supported by DFG, SFBTR22, project A07).
induce within an epithelial cell is an important step on the way
to understand the pathogenesis of epithelial diseases and their
selected puBlications
most important findings
treatment. Recent findings revealed FoxO, a member of the
Hagner S, Harb H, Zhao M, Stein K, Holst O, Ege E, Mayer
Allergy and asthma: Different mode of action of allergy-
forkhead box family of transcription factors, as a potential key
M, Matthes J, Bauer J, von Mutius E, Renz H, Heine* H, Pfef-
protective cowshed bacteria
regulator of stress in cellular processes. In contrast to humans,
ferle* PI, Garn* H (* equal contribution). Farm-derived Gram
The incidence of allergic disorders has strongly increased in
who possess four FoxO proteins, the fruit fly Drosophila melano-
positive bacterium Staphylococcus sciuri W620 abolishes asth-
industrial countries in the last decades. This is thought to be
gaster has only one homologue, making Drosophila an excel-
ma phenotype in HDM and OVA treated mice. Allergy. 2013 Feb
due, among others, to lesser exposure to microbes/microbial
lent model to analyze its functions.
1. doi: 10.1111/all.12094. [Epub ahead of print]
compounds as it is the case in urban environment and western
We succeeded in identifying FoxO as a central mediator, which
lifestyle. Evidence from several epidemiological studies show
is activated by various environmental stressors. Furthermore, this
Warmbold C, Uliczka K, Rus F, Suck R, Ulmer AJ, Petersen
an inverse relationship between farming environment in early
stress responsive function of FoxO is highly conserved in the
A, Silverman N, Heine H, Roeder T. Der p1-induced immune
childhood and the occurrence of allergic diseases in later life.
respiratory epithelia of flies and mammals (Fig.2).
Recently, we demonstrated that the Gram+ cowshed isolate
responses in the fruit fly Drosophila melanogaster. J Immunol
2013; 190, 1:366-371.
Lactococcus lactis G121 was able to prevent allergic immune
responses in a mouse model of acute allergic inflammation and
Heine H. TLRs, NLRs and RLRs: Innate sensors and their impact
this protection was due to the induction of a Th1 shift. Another
on allergic diseases – A current view (review). Immunology Let-
cowshed isolate, Staphylococcus sciuri W620, has also been
ters 2011; 139: 14-24.
shown to be protective in vivo using OVA and HDM mouse mod-
internal and external collaborations
els. However, the mechanism of cell activation by both bacteria
differs especially with regard to their ability to induce a Th1-po-
Figure 1: S. sciuri W610 induces the release of IFN-γ in human moDC /
larizing immune response. In vitro data of transiently transfected
T cell coculture experiments.
HEK293 cells showed that both bacteria activate cells via the
Figure 2: Oxidative stress induces the translocation of FoxO in Droso-
Otto Holst, Arnd Petersen, Uta Jappe, Helmut Haas, Nicolas
phila as well as the translocation of FoxO proteins in human epithelial
Gisch, Helmut Brade
airway cells.
Thomas Roeder, CAU Kiel
intracellular Nod2 receptor but only S. sciuri W620 displayed a
of IFN-γ in the coculture experiments. Further supporting the dif-
strong TLR2-dependent activity. Stimulation of human dendritic
ference in the mode of cell activation by both bacteria we also
An important line of defense against microbial invasion in
Peter König, University Lübeck
cells (DCs) with both bacteria led to a substantial upregulation
found an upregulation of IFN-β mRNA as well as IL-1β mRNA
Drosophila is the synthesis of antimicrobial peptides (AMPs).
Harald Renz, Holger Garn, Petra Pfefferle, University Marburg
of CD40, CD80, CD83 and MHCII on the cell surface. However,
and cytokine release only in L. lactis-treated DCs. S. sciuri W620
The induction of AMP gene expression was thought to be com-
Carsten Schmidt-Weber, TU and Helmholtz Center München
only L. lactis G121 induced a characteristic Th1-polarizing pro-
treatment did not induce IFN-β mRNA and although IL-1β mRNA
pletely dependent on NF-κB signaling in response to pathogen
file in DCs such as the mRNA upregulation of the costimulatory
was upregulated, no protein was detectable in the supernatant.
challenge. We demonstrate that stress conditions are also ca-
Grant support
Th1-associated molecule Delta4 and downregulation of the Th2-
Overall, we found that the cowshed bacteria L. lactis G121 as
pable to induce an immune response in a pathogen- and NF-
SFB/TR22, project A02
associated Jagged-1-mRNA as well as the release of IL-12p70
well as S. sciuri W620 were allergy-protective; however, the
κB-independent manner but under the participation of FoxO.
BMBF DZL, disease area AA2.2
and TNF-α. Except for low amounts of TNF-α in the supernatants
mechanisms seem to differ profoundly. While L. lactis G121 in-
On the basis of current findings we conclude that as a result
a stimulation of DCs with S. sciuri W620 did not induce any of
duced a characteristic Th1-polarizing immune response in DCs
of starvation, FoxO is activated via the insulin pathway and
these Th1 modulators. Despite the lack of induction IL-12p70 re-
S. sciuri W620 showed a rather generalized suppression of T
can induce AMP gene expression independently of the NF-
lease, coculture experiments with human DCs and naïve CD4+
cell activation in vivo (supported by DFG, SFB/TR22, project A2).
κB-regulated innate immune pathways, thus revealing not only
T-cells revealed that also S. sciuri W620 led to a robust release
65
Ruth Schmitz-Streit, CAU Kiel
a new mechanism to activate a basic immune response but
of the Th1 cytokine IFN-γ (Fig.1). The explanation for this effect
Characterization of the transcription factor FoxO as a stress
also a new connection between metabolism and immunity. An-
might be that albeit its failure to induce IL-12p35 mRNA expres-
mediator in the respiratory epithelium of the model organism
other immune activating inducer is oxidative stress. The oxida-
sion, S. sciuri W620 does induce IL-12p40 as well as IL-23p19
Drosophila melanogaster (in collaboration with Prof. Thomas
tive stress mediated AMP activation seems to be regulated by
mRNA expression. Thus, IL-23 could replace IL-12 in the induction
Roeder, CAU Kiel)
other signaling mechanisms than the Insulin/IGF pathway and
Priority Research Area Asthma & Allergy • Innate Immunity
Prof. Dr. Holger Heine
Mouse Models in Asthma
airway inflammation
Linda Lang • Lars Lunding
asthma phenotypes
lung function
animal models
Dr. Michael Wegmann
MISSION
melanocortin receptors is significantly altered in mice with
To establish mouse models of different asthma phenotypes 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 these respective diseases.
experimental asthma: While healthy animals exclusively express
67
the melanocortin receptor 5 (MC5-R) in the airway epithelium,
animals
with
experimental
asthma
revealed
increased
most important findings
By using these mouse models it became clear that the TH1/TH2
in infiltrating eosinophils. Furthermore, they also express MC1-R
Allergic bronchial asthma is a complex syndrome comprising
balance is critically important for the formation and initiation
in infiltrating neutrophils and fibroblasts. The altered expression
a variety of different phenotypes, which differ in points of
of the allergic immune response, but that it is not the only level
profile of α-MSH and its respective receptors by structural
severity and course of disease, corticosteroid responsiveness,
regulating airway inflammation. The balance between pro-
cells of the airways and infiltrating cells of the immune system
and airway inflammation. The patho-mechamisms underlying
inflammatory mediators (e.. g. IL-1β, IL-6, NGFs, VEGF) and anti-
suggest a regulatory role for this mediator-receptor-axis in
the formation of these asthma phenotypes remains entirely
inflammatory factors (e. g. IL-10, TGF-β) appears to be another
asthma pathogenesis and will be investigated in the next future.
enigmatic, but imply an imbalance of different regulatory levels
regulatory level that has marked impact on the maintenance
during initiation, progression and chronification of allergic
of airway inflammation and, thus, chronification of allergic
selected puBlications
airway inflammation. Elucidating these processes is the key to
asthma. Since viral infections of the upper respiratory tract
Wegmann M, Lunding L, Orinska Z, Wong DM, Manz RA,
OVA were exposed to OVA-aerosl, while the midexpiratory airflow
understanding the pathogenesis of allergic asthma.
have been identified to be the major cause for acute asthma
Fehrenbach H. Long-term Bortezomib treatment reduces
(EF50) was continually assessed.
exacerbations, which in turn are the main risk factor for the
allergen-specific IgE but fails to ameliorate chronic asthma in
One of these regulatory levels is the balance between T helper
progression towards severe asthma phenotypes, we further
mice. Int Arch Allergy Clin Immunol 2012; 158: 43-53.
1 (TH1) and TH2 type immune responses, which bases on the
established a mouse model for asthma exacerbation. By local
fact that TH1 and TH2 polarized immune responses control
application of viral Toll-like receptor ligands to mice already
Wegmann M. The role of neutrophils and TH17 cells in the
each other. The last two decades figured out that in allergic
suffering from acute experimental asthma, we mimicked viral
immunopathology of severe asthma. Anti-Inflammatory & Anti-
diseases such as bronchial this balance is shifted towards
infection and set a strong pro-inflammatory stimulus. This indeed
Allergy Agents in Medicinal Chemistry 2011;10(6):427-41.
a predominance of TH2 cells, which are the main leukocyte
led to induced/provoked exacerbation of acute allergic asthma
subpopulation orchestrating the allergic immune response –
as characterized by pronounced infiltration of neutrophils and
Wegmann M. Targeting Eosinophil’s Biology in Asthma Therapy.
and mouse models of experimental asthma contributed
eosinophils into the airways, increased mucus production,
Am J Respir Cell Mol Biol. 2011;45(4):667-74.
substantially to that knowledge.
and markedly impaired lung function. Interestingly, this was
Figure 1: Acute broncho-obstruction after antigen-aerosol inhalation.
Mice sensitized to and long-term challenged with the model-allergen
associated with enhanced production not only of typical TH2
internal and external collaborations
Figure 2: Lung function and airway inflammation in mice with acute
However, the “classic mouse models of experimental asthma”
type cytokines (e.g. IL-4, IL-5, IL-13) but also of generally pro-
At the Research Center Borstel we cooperate with the Divisions
asthma exacerbation. Mice were sensitized to and challenged with
were limited in reflecting the human disease phenotype due
inflammatory cytokines (e.g. IL-1β, IL-6, TNF-α), neutrophilatactic
of Biochemical Immunology, Cellular Microbiology, Experimental
to their acute character or their artificial route of sensitization.
mediators (e.g. IL-8) and increased infiltration of activated TH17
Pneumology, Fluorescence Cytometry, Immune Cell Analytics,
Thus, we have improved the “classic mouse models”: On the
cells. How this process is mediated and whether TH17 cells play
Infection Immunology, and Invertebrate Models. External
ased leukocytic influx into the airway as assessed by broncho-alveolar
one hand models are now available that use adjuvant-free
a central role in that will be subject to future research.
cooperations have been established with Prof. Rudolph Manz
lavage (BAL) (2A) associated with increased airway hyperresponsive-
from the Institute of Systemic Inflammation Research (ISEF) at the
ness as assessed by metacholin (MCh) provocation testing with headout plethysmography (2B). The dose of MCh that is required to reduce
sensitization and on the other hand we have established
a mouse model of chronic experimental asthma. This is
While this project focuses on the role pro-inflammatory factors
University Lübeck, and Porf. Markus Böhm from the Department
characterized by chronic inflammation of the airway and signs
in asthma pathogenesis, another project addresses potentially
of Dermatology at the Hospital of the University Münster.
of airway remodeling (e. g. subepithelial fibrosis, smooth
counterbalancing anti-inflammatory mediators. One candidate
muscle hyperplasia, goblet cell metaplasia) and displays
could be the α-melanocyte stimulating hormone (α-MSH), a
Grant support
for the first time acute broncho-obstruction in response to
neuro-peptide hormone that was originally found in the pituary
BMBF DZL AA 2.1
allergen inhalation. While these models aim to mimic the
gland and stimulates the tanning reaction of the skin in response
EXC 306 STI
phenotype of eosinophil-dominated allergic asthma, we
to UV-irradiation. We found that this growth factor is produced
further established a mouse model of neutrophil-dominated
by the airway epithelium in response to allergic inflammation
asthma, which displays increased numbers of infiltrating TH17
and that the α-MSH levels detected in BAL fluid correlate with
cells and high levels of interleukin (IL) 17 in broncho-alveolar
the degree and duration of the inflammatory response in the
lavage fluid (BAL).
airways. Furthermore, the expression pattern of its respective
the model-allergen OVA to induce experimental asthma (OVA).
Subsequent local application of the TLR-3 ligand poly (I:C) (OVA + pIC)
induced acute exacerbation of the disease as characterized by incre-
the midexpiratory airflow (EF50) of about 50 % is expressed as MCh50
and indicates responsiveness of the airways.
Priority Research Area Asthma & Allergy • Mouse Models in Asthma
expression of MC5-R not only in the airway epithelium but also
Barrier-Integrity
Inflammation & Sepsis
Dr. Heike Dombrowsky • Tom Räwel • Jürgen Sarau • Yuk Lung Wong
Intestine & Barriers
Microcirculation & Fluid Balance
Molecular Mechanisms
Dr. Ingmar Lautenschläger
selected puBlication
the interaction of microbiocides and bacterial remnants in the
In septic and other intensive care patients, mesenteric microcirculation and intestinal barrier function are critical for survival yet very difficult to maintain. Inflammation induces endothelial and
epithelial barrier dysfunction, ultimately leading to gut failure, loss of fluid and protein from the
circulation and translocation of bacteria and remnants. Our goals are to explore the molecular
mechanisms involved and to identify relevant therapeutic targets.
Dombrowsky H , Lautenschläger I, Zehethofer N, Lindner B,
intestine.
most important findings
We have improved the technology of the isolated perfused rat
Schultz H, Uhlig S, Frerichs I, Weiler N. Ingestion of n-3 fatty
acids augments basal and platelet activating factor-induced
We thank PD Dr. T. Goldmann and Dr. H. Schultz (Division of Cli-
permeability to dextran in the rat mesenteric vascular bed.
nical and Experimental Pathology, Research Center Borstel) for
JOURNAL OF NUTRITION 2011 Sep;141(9):1635-1642).
the cooperation and help with the histopathological analysis.
ced vascular permeability as a side effect of fish oil supple-
internal and external collaborations
Grant support
mentation. Furthermore, they suggest – in line with the above
We collaborate extensively and discuss all projects with Prof.
Medical Faculty of the Christian-Albrechts-University, Kiel
small bowel model and further adjusted it to the mouse ana-
mentioned findings – that mediators other than eicosanoids
N. Weiler and Prof. I. Frerichs (working group “Intensive Care
tomy and physiology. It enables us for the first time to analyze
are responsible for the immediate vasoconstriction and edema
Medicine”, Department of Anesthesiology and Intensive Care
Leibniz-Vorhaben im wettbewerblichen Verfahren im Rahmen
the intestinal pathophysiology, in particular the intestinal micro-
formation in response to PAF and open up the possibility of
Medicine, University Medical Center Schleswig-Holstein, Cam-
des Paktes für Forschung und Innovation
circulation and its barriers in detail, in comparison of different
novel therapeutic approaches.
pus Kiel) and Prof. S. Uhlig (Institute of Pharmacology and Toxi-
species and in a compartment related fashion. Using this model we have shown that platelet activating factor (PAF) as an
archetypical mediator of inflammation induces vascular and
barrier dysfunction as well as changes in fluid homeostasis
cology, Medical Faculty of RWTH Aachen University).
Inflammation induced by the
platelet activating factor (PAF)
likely seen in acute intestinal failure.
the effects of different intracellular metabolites and mediators
rendering the following results: First, thromboxane and leukotrienes seem not to play a major role in PAF-mediated intestinal
microcirculatory disturbances and, second, arachidonic acid
more than 40 different molecular species, demonstrating individuality of phospholipid classes composition and high flexibility
G-Prot.
lipids as well as of intracellular kinases. They led to the hy-
barrier dysfunction. We consequently moved on and analyzed
position by HPLC-MS. The state of the art technique identified
PAF-R
ment with antagonists of e. g. the eicosanoids, the sphingo-
responsible for the microcirculatory disturbances and for the
chemistry, Research Center Borstel) we have examined the
influence of dietary n-3 FA on the intestinal phospholipid com-
Our studies of the intestinal PAF effects included the pre-treat-
pothesis that there were different cellular signaling pathways
In cooperation with PD Dr. B. Lindner (Division of Immuno-
of the intestinal lipid pool.
PLA2
AC-inhibition
Arachidonic acid
cAMP-depletion
Thromboxane,
Leukotrienes,...
PD Dr. A. Frey (Division of Mucosal Immunology and Diagnostics, Research Center Borstel) cooperates with us to explore
the processing of antigens at mucosal surfaces. In these experiments, the isolated intestine provides an environment for
the reproducible application of antigens and identification of
routes across the intestinal epithelium.
metabolites are only partially responsible for PAF-mediated
gut injury. Furthermore, the anti-inflammatory agent quinidine
as well as elevated intracellular cAMP are able to protect the
gut from PAF, probably by ion channel regulation and/or a calcium dependent pathway (manuscript in preparation).
The goal of the collaboration with Dr. C.E. Boyle (Department of
Dysperfusion and Hyperpermeability
Figure 1: Proposed relevant mechanisms of platelet activating factor
(PAF) in the intestine“
We published our studies on the effects of dietary n-3 fatty
Medical Microbiology, Virology and Hygiene, UKE, Hamburg)
and Prof. G. Graßl (Models of Inflammation, Research Center
Borstel) is the exploration of the interaction of yersinia and
salmonella with the intestinal mucosa. With a laboratory fulfilling the S2/L2 criteria we are able to safely conduct isolated
acids (FA) of PAF induced vasoconstriction and hyperperme-
To further demonstrate the utility of our isolated organ model
perfusion experiments with pathogenic strains and genetically
ability. Despite incorporation of n-3 FA into intestinal phos-
we established a novel ex vivo infection model of Yersinia and
modified organisms. These experiments will elucidate routes of
pholipids and PAF-induced eicosanoid generation we found
Salmonella. In this collaboration project we aimed to charac-
Yersinia invasion and thus provide insight in the pathophysiolo-
no alleviation of vasoconstriction and even elevated intestinal
terize the interactions of these pathogens with the epithelium
gy of the disease.
permeability in the isolated intestine. These data indicate that
as well as their effects on intestinal fluid balance (manuscript
the beneficial antiinflammatory effects of n-3 FA in critically ill
in preparation).
patients need to be balanced against the possibility of enhan-
Together with Prof. K. Brandenburg and Dr. L. Heinbockel (Division of Biophysics, Research Center Borstel) we investigate
69
Priority Research Area Asthma & Allergy • Barrier-Integrity
MISSION
Structural Biochemistry
Lipopolysaccharide
glycolipids
polysaccharides
Dr. Katarzyna Duda • Dr. Uwe Mamat • PD Dr. Sven Müller-Loennies • Dr. Young-Hwa Song • Anna Czabanska • Sandra Kurdewan • Olga Neiwert • Ute Agge • Rainer Bartels • Helga Bartels Petra Behrens
Sylvia Düpow • Regina Engel • Dörte Grella • Volker Grote • Katharina Jakob • Gudrun Lehwark-Yvetot
Veronika Susott • Christine Schneider • Kathleen Wilke
hygiene hypothesis
Prof. Dr. Otto Holst
MISSION
directed mutagenesis experiments determined critical roles
Fischer K, Vinogradov E, Lindner B, Heine H, Holst
The Division of Structural Biochemistry is devoted to the isolation, structural analysis and molecular
biology of compounds from natural material, i.e. carbohydrates and lipids from microbes and environmental samples.
for glycine 30 and glutamate 31 in Kdo transfer. This data
O. The structure of the extracellular teichoic acids from
represent the structural basis of a critical reaction in LPS
the allergy-protective bacterium Lactococcus lactis G121.
biosynthesis and allows the development of a detailed model
BIOLOGICAL CHEMISTRY 2012;393:749-755.
71
most important findings
outer membrane identified sphingolipids as major lipid class,
The Gram-positive cowshed bacterium Lactococcus lactis
together with ornithine-containing lipids (OL) and etherlipids.
G121 modulates the immune system resulting in allergy
protection. Lipoteichoic acids (LTA) represent one major Gram-
internal and external collaborations
The farm isolate and Gram-positive bacterium Lactococcus
Internally, we are collaborating with the Divisions of Innate
More than 50 structural variants within these three classes
lactis G121 was shown to prevent mice from ovalbumin-induced
Immunity (H. Heine), Cellular Microbiology (U. Schaible), and
were found which possessed several interesting properties
asthma. The isolation and characterization of cell envelope
Clinical and Experimental Pathology (E. Vollmer, T. Goldmann).
positive cell envelope component which is considered a
concerning LPS replacement, mediators in myxobacterial
constituents represents a prerequisite for the understanding of
Externally, we are collaborating with the Dipartimento di
pathogen-associated molecular pattern. The isolation as well
differentiation as well as potential bioactive properties. The
the molecular mechanisms of such allergy-protective properties.
Chimica e Biochimica, Universitá di Napoli Federico II, Naples,
as structural and functional analyses of the LTA of L. lactis
sphingolipids with the basic structure C9-methyl-C20-sphingosine
The structural analyses of the extracellular teichoic acid (EC
Italy (A. Molinaro, C. De Castro), Experimental Pneumology,
G 121 were performed. Butan-1-ol extraction and purification
possessed a C9-methylation which is common to fungi but have
TA) from L. lactis G121 were performed. Extraction with 0.9 %
Ruhr-University
by hydrophobic interaction chromatography yielded pure LTA.
not been found in bacteria before. They may have a function
saline yielded a crude teichoic acid fraction which was purified
Children Hospital, LMU, Munich, Germany (E. von Mutius),
Structural investigations utilized chemical analytical methods,
in myxobacterial development. The OL, also identified in
by size exclusion chromatography on Biogel P60 and P10
Chair of Animal Hygiene, TUM, Munich, Germany, Institute
nuclear magnetic resonance spectroscopy and high resolution
myxobacteria for the first time, contained acyloxyacyl residues
matrices. Compositional analyses, high resolution electrospray
for Laboratory Medicine and Pathobiochemistry, University of
electrospray ionization Fourier-transformed ion cyclotron mass
which are also characteristic for LPS and might replace these
ionization Fourier-transformed ion cyclotron mass spectrometry
Marburg, Germany (H. Renz), Haartman Institute, Department
spectrometry. The LTA isolate contained a heterogeneous
molecules in certain functions. The etherlipids are considered
and nuclear magnetic resonance spectroscopy were performed
of Bacteriology and Immunology, Helsinki, Finland (M. Skurnik),
mixture of molecules composed of a 1,3 poly(glycerol
biomarkers in myxobacterial development.
for structural elucidation. The EC TA was a poly(glycosylglycerol
Institute of Microbiology, University of Silesia, Katowice,
Figure 2: The crystal structure
phosphate) molecule comprising a repeating unit of -6)-[β-D-
Poland (J. Radziejewska-Lebrecht), Department of Genetics
of the free WaaA from Aquifex
Glcp-(1→3)-][α-D-GlcpNAc-(1→4)-]α-D-GalpNAc-(1→3)-β-D-
and Microbiology, Maria Curie-Skłodowska University, Lublin,
aeolicus.
GlcpNAc-(1→2)-glycerol-(1-P-.
Poland (A. Choma), Institute of Nanobiotechnology, University
phosphate) backbone that was randomly substituted at C-2
by D-alanine and α-D-Galp (see structure below). The lipid
anchor constituents were kojibiose linked to a heterogeneous
Bochum,
Germany
(A.
Bufe),
University
of Natural Resources and Life Sciences, Vienna, Austria (C.
diglyceride comprising a total of six different fatty acid
compositions. This LTA possessed TLR2- and TLR4-independent
selected puBlications
´
Schäffer), Faculty of Biology, University of Bielefeld, Germany
cytokine-inducing activity in human mononuclear cells.
Theilacker C, Kaczynski Z, Kropec A, Sava I, Ye L, Bychowska
(K. Niehaus).
A, Holst O, Huebner J. Serodiversity of opsonic antibodies
against Enterococcus faecalis-glycans of the cell wall revisited.
Grant support
PLoS ONE 2011;e17839.
Our research has been supported by the DFG (SFB-TR 22, project
A2, FOR 585), the BMBF (DZL), the European Union (GALTRAIN)
Keck M, Gisch N, Moll H, Vorhölter F-J, Gerth K, Kahmann
U, Lissel M, Lindner B, Niehaus K, Holst O. Unusual outer
WaaA is a key enzyme in the biosynthesis of LPS, and is
Figure 1: Structure of the lipoteichoic acid from L. lactis G121.
membrane lipid composition of the Gram-negative, lipopoly-
embedded in the cytoplasmic face of the inner membrane. It
saccharide-lacking myxobacterium Sorangium cellulosum
catalyzes the transfer of 3-deoxy-D-manno-oct-2-ulosonic acid
So ce56. THE JOURNAL OF BIOLOGICAL CHEMISTRY 2011;
(Kdo) to the lipid A precursor of LPS. The crystal structures
286:12850-12859.
of the free (structure see below) and CMP-bound forms of
Sorangium cellulosumSo ce56 is a Gram-negative myxobacterium
WaaA from Aquifex aeolicus, an ancient Gram-negative
Schmidt H, Hansen G, Singh S, Hanuszkiewicz A, Lindner
that bears the largest bacterial genome sequenced so far, with
hyperthermophile,
structures
B, Fukase K, Woodard RW, Holst O, Hilgenfeld R, Mamat
some 10,000 genes. Analysis of this genome revealed that
revealed details of the CMP-binding site and implicated a
U, Mesters JR. Structural and mechanistic analysis of the
several genes coding for the biosynthesis of lipopolysaccharides
unique sequence motif (GGS/TX5GXNXLE) in Kdo binding.
membrane-embedded glycosyltransferase WaaA required for
(LPS) were missing. Biochemical analysis confirmed this in
A cluster of highly conserved amino acid residues was
lipopolysaccharide synthesis. PROCEEDINGS OF THE NATIONAL
giving no evidence for the presence of LPS in the membranes
found representing the putative membrane-attachment and
ACADEMY OF SCIENCES 2012;109:6253–6258.
of So ce56. Analysis of the complete lipid composition of the
acceptor-substrate WaaA binding site. A number of site-
were
characterized
which
and industry.
Priority Research Area Asthma & Allergy • Structural Biochemistry
of WaaA’s catalytic mechanism.
Cellular Allergology
Anti-inflammatory
worm factors
Beate Höschler • Dr. Arne Homann • Katrin Knuhr • Kristina Langhans • Sandra Nyenhuis • Dr. Natalie
Reimers • Dr. Gabriele Schramm
In vitro worm culture as
a universal tool
PD Dr. Helmut Haas
vitro cultured worms represents a remarkable complement to
J. Müller-Dieckmann, EMBL, Hamburg
To identify and characterize anti-inflammatory molecules from parasitic worms, namely Schistosoma
mansoni, and to employ them for protection against asthma, allergies and autoimmune disorders. Our
motto: “Learn from the Worm!”
conventional HTS. As a “reductionist animal model” it allows
C.G. Grevelding, Universität, Giessen
studying (e.g. conserved) biological events in a relevant in vivo
M. J. Doenhoff, University of Nottingham, UK
context ensuring relevant results. Thus, HTS on in vitro cultured
A. Cooke, University of Cambridge, Cambridge, UK
worms will compress and accelerate the preclinical part of
F.H. Falcone, University of Nottingham, UK
In vitro culture of schistosomes as a universal tool, e. g. for
the drug discovery process by making various procedures
M. Böhm, Universität Münster
Omega-1, a Schistosoma egg glycoprotein, requires both
high through-put compound screening (HTS)
superfluous. This will save mammalian animal lives and
E.J. Pearce, Washington University School of Medicine; St. Louis,
its glycan elements and its RNase activity to condition
Schistosoma mansoni is a parasitic worm that kills about
drastically reduce the cost for launching a new drug.
USA
dendritic cells (DC) for Th2 priming
200 000 people per year in sub-Saharan Africa. Since
Site-directed mutagenesis of omega-1, a glycosylated T2
resistances to praziquantel, the present drug of choice, are
selected puBlications
Grant support
ribonuclease (RNase) secreted from Schistosoma mansoni
being observed, the search for alternative drugs against
Everts B, Hussaarts L, Driessen NN, Meevissen MH, Schramm
BMBF 0315277 „In vitro culture of Schistosoma mansoni – Culture
eggs, revealed that both the glycosylation and the RNase
schistosomiasis is urgently needed. As our lab is the only lab
G, van der Ham AJ, van der HB, Scholzen T, Burgdorf S, Mohrs
dish instead of mammalian host“, September 2008 – February
activity are essential for this molecule to condition DC for
worldwide where S. mansoni is grown in vitro from the larval
M, Pearce EJ, Hokke CH, Haas H, Smits HH, Yazdanbakhsh
2013, 1 post-doc, 1 technician
Th2 polarization. Morphologically, omega-1 was found to be
to the adult stage including egg deposition, we have adapted
M. Schistosome-derived omega-1 drives Th2 polarization by
EU-FP7, grant agreement no. 242107 „The targeted development
bound and taken up by DC (Fig. 1). This process depends
our protocol to high through-put screening (HTS) as a tool for
suppressing protein synthesis following internalization by the
of a new generation Vaccine for Schistosomiasis” (TheSchistoVac;
on the presence of glycans on omega-1 and the mannose
identifying new drugs against the killer. In that context, the
mannose receptor. J Exp Med 2012; 209:1753-67, S1.
work package 5), March 2010 – February 2014, 1 Post-Doc
receptor on DC, respectively. Functionally, omega-1 impairs
in vitro culture was adapted to 384-well plates including the
protein synthesis by degrading both ribosomal and messenger
presence of DMSO in the culture medium. The usefulness and
Meevissen MH, Driessen NN, Smits HH, Versteegh R,
excretory/secretory protein from Schistosoma mansoni eggs”,
RNA. Thus, omega-1 acts via a pathway involving mannose
reliability of our procedure was unequivocally demonstrated
Van Vliet SJ, van KY, Schramm G, Deelder AM, Haas H,
October 2012 – September 2015, 1 PhD student
receptor and interference with protein synthesis to condition
by a “blinded” study using various test compounds with
Yazdanbakhsh M, Hokke CH. Specific glycan elements
DC for Th2 priming.
differing schistosomacidal capacities (Fig. 2).
determine differential binding of individual egg glycoproteins of
most important findings
DFG SCHR 608/4-1 „Immunomodulation by IPSE/alpha-1, an
the human parasite Schistosoma mansoni by host C-type lectin
receptors. Int J Parasitol 2012; 42:269-77.
Kaur I, Schramm G, Everts B, Scholzen T, Kindle KB, Beetz
C, Montiel-Duarte C, Blindow S, Jones AT, Haas H, Stolnik
S, Heery DM, Falcone FH. Interleukin-4-inducing principle from
Schistosoma mansoni eggs contains a functional C-terminal
nuclear localization signal necessary for nuclear translocation
Figure 2: Effect of praziquantel on schistosome larvae cultured 10
in mammalian cells but not for its uptake. Infect Immun
days in vitro. Normal larval development under medium control con-
2011;79:1779-88.
ditions (left). In the presence of praziquantel (10 µM; right) the larvae
are dead and shrunk.
internal and external collaborations
Internal:
Notably, beyond the parasite-specific application, the in vitro
Thomas Scholzen
culture of worms offers a highly attractive tool for compound
Andreas Frey
screening in general. Up to now, most conventional HTS
Guntram Grassl
procedures are based on biochemical assays focusing on the
Holger Heine
interaction between single isolated molecules in an artificial
Norbert Reiling
context. This, however, can only distantly reflect what is going
External:
on in the complex in vivo context explaining a high rate of
Maria Yazdanbakhsh, C.H. Hokke, Leiden University Medical
microscopical section through life cells (Th. Scholzen, RC-Borstel); FITC-
biologically irrelevant results and, consequently, the high
Center, Leiden, NL
labeled omega-1: green, nucleus: blue (lysosomes: red).
cost of conventional HTS procedures. In contrast, HTS on in
M. Sattler, Technische Universität, München
Figure 1: Omega-1 is taken up by dendritic cells. Laser scanning
73
Priority Research Area Asthma & Allergy • Cellular Allergology
MISSION
Clinical and Molecular Allergology
from clinic
to research
Marisa Böttger •Gerda Finnern • Dr. Skadi Kull • Maren Hohn • Dr. Sandra Minge • Prof. Dr. Arnd Petersen • Sandra Rennert • Simone Ross • Dr. Frauke Schocker • Daniela Warneke • Lisa BleSS • Arne Cernik
Caroline Morawski • Janna Müller • Daniel Rosero
and back to clinic
Prof. Dr. Uta Jappe
consumption of red meat. In a prospective multicenter study
internal and external collaborations
• 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
we investigated the prevalence and clinical relevance of anti-
Inhouse Divisions
alpha-GAL-IgE in German patients with meat/innard allergy (30
Mucosal Immunol. & Diagnostics; Clin. and Exp. Pathology, Exp.
min to 8 h after ingestion) and severe urticaria/anaphylaxis due
Pneumology, Innate Immunity; Structural Biochemistry, Biophysics
to unknown causes. Sera were analyzed by immunoblotting and
National
compared with the alpha-GAL ImmunoCAP (Platts-Mills, Virginia,
O. Janßen, Univ. of Kiel: 2D DIGE and mass fingerprinting; A.
USA). The ImmunoCAP was shown to be more specific for meat/
Kromminga, IPM Biotech, Hamburg: IgE-binding epitopes on
innard anaphylaxis than the cetuximab based immunoblot.
biological; H. Notbohm, University of Lübeck: Secondary structure
Thus, alpha-GAL seems to be strongly associated with severe
of allergens; J. Saloga, University of Mainz: Modulation of the
and partly delayed allergic reactions. Regarding urticaria
human immune response using allergens on APCs; S. Schillberg,
Translational aspects
• Ascertaining the acquisition of well-defined patient subgroups
• Establishing registers for allergens
• Performing provocation tests
by close attachment to two allergy outpatient clinics, headed by Prof. Dr. Jappe
patients, our immunoblot is by far more sensitive, indicating a
Fraunhofer Inst., Aachen: Expression of allergens in tobacco; C.
most important findings
peanuts at first exposure. Breast milk samples were collected
broader spectrum of epitopes. These assays are useful tools for
Traidl-Hoffmann (ZAUM, Munich): Allergen uptake via human skin;
Two new peanut allergens
from 32 lactating women prior and after consuming roasted
risk management of target treatments in the future.
B. Przybilla, (LMU, Munich): Allergens in delayed anaphylaxis
Peanut is one of the most hazardous food allergen sources.
peanuts at different time points. The samples were analysed
Because of the high lipid content of peanut, allergens might be
by SDS-PAGE, immunoblotting and by a sandwich ELISA using
lost in the aqueous diagnostic extract. Therefore, the focus is
the commercially available Neogen Veratox ELISA® and an
on the purification of hydrophobic allergens to determine their
ELISA against digestion-resistant Ara h 2 (DRP-Ara h 2), a marker
relevance. In 2012 we detected IgE-reactive proteins of about 6
allergen of peanut for severe allergic reactions. Only by use of
kDa in the effluent. Allergenicity was confirmed by IgE detection
the latter assay DRP-Ara h 2 was detected in 25 % of the samples
(immunoblot) and by basophil activation test (CD203c). Protein
ranging from 50 to 2500 ng per ml breast milk, while the two
sequencing and homology search identified the isolated
other tests were too low in sensitivity. These results indicate
components as defensins. The two peanut defensins were
that peanut protein is secrected into breast milk after maternal
accepted by the IUIS allergen nomenclature subcommittee as
peanut ingestion.
two novel peanut allergens, Ara h 12 and Ara h 13.
IgE to certain epitopes as biomarker for risk assessment
Hypersensitivity reactions to biologicals seem to be increasing.
International
J. Baumert, University of Nebraska, USA): Detection of peanut
Anti-α-GAL-IgE
Galactose-α1,3-Galactose = α-GAL
Meat allergy:
Center, Amsterdam: Stimulation of the innate immune system;
Anti-α-GAL-IgE bind to
several different proteins of
non-primate mammals
Parasites
(Arthropods, Helminths)
Ticks, Nematodes
Delayed anaphylaxis
Development of symptoms
often at night
No association with asthma
Glycosylation of
recombinant therapeutic
molecules:
safety risk for severe
allergic reactions?
Possible wrong diagnosis:
Idiopathic anaphylaxis
Conventional commercially
available diagnostic extracts
mostly insufficient
Studies
M. Gavrovic-Jankulovic, University of Belgrade, Serbia: structural
Investigation of the
prevalence and clinical
relevance of
analysis of allergens; A. Jacquet, Chulalongkorn University,
α-GAL-IgE
in A. German patients
with meat/innard allergy
and severe urticaria /
anaphylaxis
in B. patients with allergy
to biologicals
Thailand: Protein expression in Pichia pastoris; J. Lidholm,
Uppsala, Sweden: Peanut and lupine allergy; A. Mari, Rome,
Italy: Identification of food allergens considering geographical
aspects; T.M. Platts-Mills (Charlottesville, USA): cross-reactive
carbohydrate determinants; A. Sette, San Diego, USA: T cell
epitope recognition
Jappe U, Der Hautarzt 2012, modified
Grant support
The therapeutic antibodies infliximab, cetuximab, adalimumab
and rituximab have been identified as allergenic. Anaphylaxis
allergens/traces by ELISA; C. van Drunen, Academic Med.
Figure 2: Sensitization routes and clinical impact of a glycan epitope
to cetuximab, a chimeric mouse-human IgG1 monoclonal
SFB/Transregio 22 „Allergic immune responses of the lung“,
Projekt Z1 (Lindner B., Petersen A.)
antibody approved for oncological target therapy, is caused
selected puBlications
DFG Scho 828/2-1 „Untersuchung von Erdnussantigenen/-
by pre-existing IgE to the disaccharide galactose-[alpha]-
Heydenreich B, Bellinghausen I, König B, Becker WM,
allergenen in der Muttermilch nach Prozessierung an den
1,3-galactose (alpha-GAL). Therefore, an assay is needed to
Grabbe S, Petersen A,* Saloga J.* Gram-positive bacteria on
Grenzflächen und ihre Interaktion mit dem Immunsystem“
identify IgE-binding epitopes on biologicals as a prerequisite
grass pollen exhibit adjuvant activity inducing inflammatory T
DFG
for IgE detection before and during target treatment as a
cell responses. CLIN. EXP. ALLERGY 2012; 42:76–84.
Funktionsuntersuchungen von lipophilen Erdnussallergenen,
measure of risk management. We developed a sensitive and
JA1007/2-1
und
DFG
PE
491/10-1
“Struktur-und
insbesondere den Olesosinen“
specific method for the detection of anti-alpha-GAL-IgE by use
Möbs C, Ipsen H, Slotosch C, Petersen A, Würtzen PA, Hertl
Member of the German Center for Lung Research and Airway
of alpha-GAL coupled to human serum albumin and cetuximab
M, Pfützner W. Loss of allergen-specific T cell responses and
Research Center North (Jappe U., Petersen A.)
as target in an immunoblot. For detailed studies the therapeutic
prolonged synthesis of inhibitory IgG antibodies is linked to
Bundesministerium
Breast milk: Sensitization route for peanut allergy?
antibodies were separated into Fab and Fc, and the responsible
long-term efficacy of birch pollen immunotherapy. J. ALLERGY
(BMWi)
Since allergic reactions to peanut seem to occur at the first
IgE-binding epitopes seem to be located on the Fab regions.
CLIN. IMMUNOL. 2012; 130:1108–1116.
Innovationsprogramm
known ingestion, the exposure to peanut allergens must have
Interestingly, the same IgE-binding alpha-GAL epitope, which
taken place much earlier probably during lactation predisposing
is highly immunogenic because it does not occur in humans,
Jappe U. Update on meat allergy. α-Gal: a new epitope, a new
gegen therapeutische monoklonale Antikörper (Jappe U.)
babies to experience life-threatening allergic reactions to
has been associated with delayed anaphylactic reactions after
entity? DER HAUTARZT 2012; 63:299-306.
BMBF, Verbundprojekt BASALIT (Jappe U.)
Figure 1: Two new peanut allergens: Defensins – function and clinical impact
für
Wirtschaft
KF2784701AJ0:
und
Förderprogramm
Mittelstand:
Technologie
Zentrales
Entwicklung
von
Testsystemen zum Nachweis von spezifischen IgE-Antikörpern
75
Priority Research Area Asthma & Allergy • Clin. & Molecular Allergology
MISSION
MUCOSAL IMMUNOLOGY AND DIAGNOSTICS
Molecular Imaging
Mucosal Barriers
Dr. Barbara Frey • Dr. Maik Henkel • Özge Kök • Dr. Thorsten Krause • Iryna Kruse • Dr. Katrin Ramaker
Dr. Niels Röckendorf • Dr. Heike Sinnecker • Alheidis von Quast • Imke Wysokinski
High Throughput
Peptide Assays
Molecular Evolution
MISSION
they encounter the epithelial surfaces in the intestine. Such
SPP Biomedizintechnik: University of Lübeck
The Division of Mucosal Immunology and Diagnostics focusses its research on the molecular and
cellular events at the mucosal surfaces of the respiratory system and the gastrointestinal tract with
the goal to better understand, diagnose, prevent and treat inflammatory, neoplastic and allergic
disorders.
modifications and agglomerations can reduce the binding to
DZL German Center for Lung Research: LMU München
and uptake by epithelial cells in vitro. In an ex-vivo rat intestinal
University of Kiel
system we could show that transport of undestroyed particulate
Veterinary Research Institute, Brno, Czech Republic
material and macromolecules, such as proteins or complex
carbohydrates, across the healthy epithelium is a rare event.
Grant support
most important findings
of only 4400 peptides synthesized and tested, a more than
Thus, the pathological processes observed in allergic asthma
Bundesministerium für Bildung und Forschung (BMBF), grant
The activities of the Division of Mucosal Immunology and
100fold improvement in target recognition could be achieved
or food allergies most likely are caused by smaller entities such
13N10505
Diagnostics can be divided into two key projects: The
(Figure 1), rendering the resulting peptides valuable candidates
as protein fragments or peptides.
Deutsche Forschungsgemeinschaft (DFG), grant Fr 958/5-2
“Molecular Imaging Project” deals with the diagnosis of
for the specific targeting of a reporter unit to the cell surface.
University of Lübeck, grant F249607
pathological conditions at mucosal surfaces. We want to create
For the in vivo validation of such novel contrast agents we
selected puBlications
smart contrast agents and probes for in-vivo molecular imaging
have established an orthotopic tumor model which allows
Gorris HH, Bade S, Röckendorf N, Franek M, Frey A. Pushing
and in-vitro point of care diagnostics. Targeted disease areas
the detection of cancer cells by fluorescence endoscopy with
antibody-based labeling systems to higher sensitivity by linker-
include neoplasia of the colon and the lung and inflammatory
hitherto unmatched sensitivity.
assisted affinity enhancement. BIOCONJUGATE CHEMISTRY.
pulmonary diseases such as COPD and asthma.
German Center for Lung Research (BMBF) AA2.2 & COPD-2
2011; 22:1619-1624.
The “Mucosal Interface Project” explores the (immuno-)
The use of contrast agents can considerably improve the
physiologiocal status at mucosal surfaces. Here, we analyze,
Ramaker K, Röckendorf N, Frey A. Engineering contrast
performance of certain imaging modalities, such as magnetic
how the microenvironment of mucosal surfaces such as
agents for gastrointestinal Magnetic Particle Imaging: the
resonance imaging (MRI) or endoscopy. If highly specific
mucus, immunoglobulins and proteases influences processing,
biological perspective. In: TM Buzug & J Borgert (Eds): Magnetic
contrast agents that label definite target structures or molecules
transport, presentation or interception of substances that come
Particle Imaging, SPRINGER PROCEEDINGS IN PHYSICS. 2012;
are applied, the visualization of a disease marker down to the
into contact with the mucosae, and we investigate if alterations
140:205-210.
(sub)cellular level may be possible, thereby greatly enhancing
in this status occur in disease and may be employed for
the diagnostic power of these imaging applications. In general,
diagnostic purposes and therapeutic interventions.
such specific contrast agents combine a reporter unit, e. g. a
77
Röckendorf N, Borschbach M, Frey A. Molecular evolution
of peptide ligands with custom-tailored characteristics for
fluorophore or an MRI label, with a specific probe binding unit
We have established several model systems with which we
targeting of glycostructures. PLOS COMPUTATIONAL BIOLOGY.
- the “targeting ligand” - that can direct the probe to the target
can study the different aspects of relevant interactions on the
2012; 8:e1002800.
of interest. Our research goal is to provide suitable targeting
molecular, cellular and tissue-level: In vitro assays with intestinal
ligands for specific diagnostic purposes on the basis of peptide
and bronchoalveolar fluids yield information on the proteolytic
internal and external collaborations
molecules. In order to identify peptides that can act as targeting
stability / half lifes of ingested or inhaled substances and on
Inhouse:
Figure 1: Improvement of the target binding capability of peptide
ligands in a given setting we devised a special strategy for the
the potential biochemical modifications which such substances
Divisions of:
probes over 10 rounds of evolutionary optimization (Röckendorf et al.,
directed “evolution” of peptidic molecules. The methodology
undergo in this environment. Cell culture studies reveal which
Barrier Integrity
PLoS Comput Biol 2012; 8:e1002800).
combines in silico evolution with in vitro testing to quickly obtain
properties of the investigated substances are relevant for cell
Biophysics
promising candidates with desired properties. The process
binding and transport processes. Animal studies and organ
Clinical and Experimental Pathology
starts with the synthesis of a population of lead peptides which
explant models disclose the fate and final tissue distribution of
Clinical and Molecular Allergology
are tested in appropriate laboratory assays for the required
taken-up material and shed light on the molecular and cellular
properties, ranked accordingly and crossed and mutated using
events – e. g. cytokine release and immune cell activation - and
External:
a genetic algorithm to create a filial generation. With this joint
the biological consequences – e. g. immune defense or allergic
DFG-funded research cluster PARENTRY: University of Lübeck;
in silico / in vitro optimization procedure stepwise improvement
reactions – ensuing from the interaction between foreign matter
University of Hamburg; University Duisburg-Essen; MPI for
of the peptides is achieved until a ligand is obtained that
and mucosal surfaces. By in-vitro simulating the oro-gastro-
Polymer Research, Mainz
meets the requirements for the application of interest. We
intestinal passage of ingested nanoparticulate material, for
BMBF-funded research consortium OPTOPROBE: ATTO-TEC,
have employed this evolution process for the identification of
example, we found that, depending on their size and surface
Siegen; University of Applied Sciences, Paderborn; GeSiM,
a peptide probe that can bind to the cell surface glycolipid
properties, nanoparticles experience different modifications by
Dresden; Karl Storz, Tuttlingen; LMTB, Berlin; R-Biopharm,
ganglioside GM1. After 10 “generations” and a total number
intestinal juices and frequently build large agglomerates before
Darmstadt
Priority Research Area Asthma & Allergy • Mucosa Immunology
PD Dr. Andreas Frey
Immunobiology
Mast cell biology
Cytokines
Andreas Baier • Dr. Rajia Bahri • Katharina Biethahn • Ann-Kathrin Brückner • Hanno Ewers • Dr. Orietta D’Orlando • Dr. Niko Föger • Dr. Nestor González-Roldán • Diego Goyeneche-Patiño • Manuel Hein
Martina Hein • André Jenckel • Frauke Koops • Rabea Langenhaun • Dr. Farhad Mirghomizadeh
Dr. Zane Orinska • Dr. Julia Polansky • Gesine Rode • Katrin Streeck • Anna Stubbe • Katrin Westphal
T cells
allergy
MISSION
reducing the clinical score and delaying disease occurrence.
Die Rolle von CD8+ regulatorischen T-Zellen und Mastzellen
Over the last few years the Labgroup Immunobiology has been focused on investigating mechanisms
controlling mast cell degranulation and CD8+ T cell effector functions.
EAE suppression is enhanced by pre-treatment of CD8(+)
bei der Induktion und Chronifizierung von allergischem Asthma
CD38(high) T cells with IL-15. These findings add evidence
FP7-Health-2007-A 201461
that the expression of ectoenzyme receptor family members
European initiative to improve knowledge, treatment and survi-
most important findings
revealed that STX11 is not only widely expressed in different
positively correlates with suppressor functions and identifies
val of haemophagocytic syndromes in children (CureHLH)
Differential regulation of mast cell degranulation versus cy-
immune cells, but also induced upon LPS or IFN-γ treatment.
CD8(+)CD38(high) T cells as potential inhibitors of excessive
FP7-Health-2007-B223151
tokine secretion by the actin regulatory proteins Coronin1a
However, Stx11 deficiency does not affect macrophage phago-
immune responses.
Understanding inflammation-associated tumorigenesis for the
and Coronin1b.
cytic function and cytokine secretion, mast cell activation, or
Föger N, Jenckel A, Orinska Z, Lee KH, Chan AC, Bulfone-Paus
antigen presentation by DCs. Instead, STX11 selectively con-
selected puBlications
S. J Exp Med. 2011, 208:1777-87.
trols lymphocyte cytotoxicity in NK and activated CD8(+) T cells
Föger N, Jenckel A, Orinska Z, Lee KH, Chan AC, Bulfone-
Mast cell (MC) activation via aggregation of the high affini-
and degranulation in neutrophils. Stx11(-/-) NK cells and CTLs
Paus S. Differential regulation of mast cell degranulation versus
ty IgE receptor (FcεRI) causes degranulation and release of
show impaired degranulation, despite a comparable activati-
cytokine secretion by the actin regulatory proteins Coronin1a
proinflammatory mediators in a process that involves the re-
on, maturation and expression of the complex-forming partners
and Coronin1b. J Exp Med. 2011, 208:1777-87.
organization of the actin cytoskeleton. However, the regula-
MUNC18-2 and VTI1B. In addition, Stx11(-/-) CTLs and NK cells
rational design of novel anti cancer therapeutic strategies
tory pathways and the molecular links between cytoskeletal
produce abnormal levels of IFN-γ. Since functional reconstituti-
D’Orlando O, Zhao F, Kasper B, Orinska Z, Müller J, Her-
changes and MC function are incompletely understood. In this
on rescues the defective phenotype of Stx11(-/-) CTLs, we sug-
mans-Borgmeyer I, Griffiths GM, Zur Stadt U, Bulfone-Paus
study, we provide genetic evidence for a critical role of the
gest a direct, specific and key role of STX11 in controlling lym-
S. Syntaxin 11 is required for NK and CD8(+) T-cell cytotoxicity
actin-regulatory proteins Coronin1a (Coro1a) and Coro1b on
phocyte cytotoxicity, cytokine production and secretion. Finally,
and neutrophil degranulation. Eur J Immunol. online 2012, print
exocytic pathways in MCs: Coro1a(-/-) bone marrow-derived
we show that these mice are a very useful tool for dissecting
2013, 43:194-208.
MCs exhibit increased FcεRI-mediated degranulation of secre-
the role of STX11 in vesicular trafficking and secretion.
Bahri R, Bollinger A, Bollinger T, Orinska Z, Bulfone-Paus
tory lysosomes but significantly reduced secretion of cytokines.
Hyperdegranulation of Coro1a(-/-) MCs is further augmented
Ectonucleotidase CD38 demarcates regulatory, memory-
S. Ectonucleotidase CD38 demarcates regulatory, memory-like
by the additional loss of Coro1b. In vivo, Coro1a(-/-)Coro1b(-/-)
like CD8+ T cells with IFN-γ-mediated suppressor activities.
CD8+ T cells with IFN-γ-mediated suppressor activities. PLoS
mice displayed enhanced passive cutaneous anaphylaxis.
Bahri R, Bollinger A, Bollinger T, Orinska Z, Bulfone-Paus S.
One. 2012,7:e45234.
Functional reconstitution assays revealed that the inhibitory
PLoS One. 2012,7:e45234.
effect of Coro1a on MC degranulation strictly correlates with
Regulatory CD8(+) T cells are critical for self-tolerance and
internal and external collaborations
cortical localization of Coro1a, requires its filamentous actin-
restricting excessive immune responses. The variety of immu-
Inger Gjertsson, University of Gothenburg, Sweden
binding activity, and is regulated by phosphorylation of Ser2 of
ne functions they fulfill, the heterogeneity of their phenotype,
Udo zur Stadt, University of Hamburg
Coro1a. Thus, coronin proteins, and in turn the actin cytoskele-
and the mechanism of action are still poorly understood.
Peter Katsikis, Dept. of Microbiology and Immunology, Drexel
ton, exhibit a functional dichotomy as differential regulators of
Here we describe that regulatory CD8(+) T cells exhibiting
University, Philadelphia, USA
immunosuppressive actions in vitro and in vivo are recogni-
Elena Vigorito, Babraham Research Center, Cambridge, England
zed as CD38(high) T cells and present in naive mice. CD38
Elena Voronov, Ron Apte, Institut of Immunology, Beer Sheva,
Syntaxin 11 is required for NK and CD8(+) T-cell cytotoxicity
is a glycosylated membrane protein with ectonucleotidase
Israel
and neutrophil degranulation.
properties. CD8(+)CD38(high) (CD44(+)CD122(+)CD62L(high))
Gillian Griffiths, Cambridge Institute for Medical Research,
D‘Orlando O, Zhao F, Kasper B, Orinska Z, Müller J, Hermans-
lymphocytes suppress CD4(+) effector T-cell proliferation in an
Addenbrooke‘s Hospital, England
Borgmeyer I, Griffiths GM, Zur Stadt U, Bulfone-Paus S. Eur J
antigen-non specific manner via IFN-γ. While direct cell-to-cell
Erietta Stelekati, Penn Institute of Immunology, Pennsylvania, USA
Immunol. online 2012, print 2013, 43:194-20.
contact is needed for this suppressor activity, it is independent
Hyu-Dong Chang, Deutsches Rheumaforschungszentrum, Berlin
Syntaxin 11 (STX11) controls vesicular trafficking and is a key
of membrane-bound TGF-β and granzyme B release. IL-15 po-
Dunja Bruder, Helmholtz Center for Infection Research
player in exocytosis. Since Stx11 mutations are causally asso-
tentiates the suppressive activity of CD8(+)CD38(high) T cells
ciated with a familial hemophagocytic lymphohistio-cytosis,
and controls their survival and expansion. In humans CD8(+)
Grant support
we wanted to clarify whether STX11 is functionally important
CD38(high) T cells inhibit CD4(+) effector T cell proliferation. In
SPP 1394 DFG/OR 101/2-1
for key immune cell populations. This was studied in primary
vivo, CD8(+)CD38(high), but not CD8(+)CD38(-) T cells mitigate
Mast cells as linkers of antiviral defence and autoimmunity
cells obtained from newly generated Stx11(-/-) mice. Our data
murine experimental autoimmune encephalomyelitis (EAE) by
DFG/TR-SFB22/A14
degranulation versus cytokine secretion in MC biology.
(INFLA-CARE)
Figure 1: Subcellular localization of the actin-regulatory protein Coronin1a and F-actin in mast cells (N.Föger)
79
Priority Research Area Asthma & Allergy • Immunobiology
Prof. Silvia Bulfone-Paus
National Reference Center for Mycobacteria
rapid detection of
multi drug resistance (MDR)
Tuberculosis
Dr. Sönke Andres • 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 Stephan • Petra Vock • Birgit Voss • Ann-Kathrin Witt
lab strengthening, external quality assurance
non tuberculous
mycobacteria
MISSION
polymorphism (RFLP) and pulsed-field gel electrophoresis are
Hepple P, Novoa-Cain J, Cheruiyot C, Richter E, Ritmeijer
The German National Reference Laboratory for mycobacteria (NRL) is part of a worldwide laboratory
network by being one of the Supranational Reference Laboratories since 1995. The NRL provides samples
and coordinates external quality assurance in the field of mycobacterial diagnostics, organized by
INSTAND e-V. Düsseldorf, since 2008. Research of the NRL focusses on the topics “evaluation of new techniques for the improvement of diagnostics”, “resistance of tuberculosis”, and “variability of clinical
mycobacterial isolates”.
technically demanding and not easy to standardize, multilocus
K. Implementation of liquid culture for tuberculosis diagnosis in
sequence typing (MLST) and mycobacterial interspaced
a remote setting: lessons learned. Int J Tuberc Lung Dis. 2011
repetitive units-variable number tandem repeats (MIRU-VNTR)
Mar;15(3):405-7.
were established and applied to characterize a set of M. avium
hominissuis strain isolated from humans (children and adults)
Hillemann D, Ruesch-Gerdes S, Boehme C, Richter E.
and from slaughter pigs. The methods were optimized in the
Rapid molecular detection of extrapulmonary tuberculosis by
sense that the loci with the highest discriminatory power were
automated GeneXpert(R) MTB/RIF system. J Clin Microbiol.
2011, 49: 1202-1205.
most important findings
MIC levels of >20.0 μg/ml (53 of 56). Three strains harbouring
chosen and used for the investigation of the M. avium strains.
Primarily the NRL is a routine laboratory to isolate mycobacteria
the mutation rpoB D516V were intermediate or susceptible at
Interestingly, it could be shown that several allelic profiles
with all recently worldwide established methods, identify
a rifampicin concentration of 20 μg/ml. The mutations katG
were present in both human and pig isolates, which clearly
internal and external collaborations
mycobacterial species with classical and molecular methods,
S315T and inhA -15C/T were most frequent in strains that were
indicates that the infections with M. avium strains are caused by
International collaborations have been established with several
and perform susceptibility testing. On a daily basis consultancy
resistant to isoniazid, at which the katG S315T mutation was
environmental strains also present in the environment and not
countries e.g. Austria, Armenia, Azerbaijan, Bosnia-Herzegowina,
is provided for laboratories and clinics concerning diagnostics
strongly associated with high-level and the inhA -15C/T mutation
specifically by highly virulent clones.
Croatia, Ghana, Kyrgyzstan, Kazakhstan, Moldova, Serbia,
and therapy of mycobacterial diseases.
was associated with low-level resistance.
In the last years new diagnostic tools for TB diagnostics
Among mycobacterial opportunistic pathogens, M. avium
selected puBlications
and the USA. The NRL consults ICRC, MSF, WHO, diagnostic
have been developed to revolutionize the rapid detection of
hominissuis is the most important for humans, but also can
Sanchez-Padilla E, Dlamini T, Ascorra A, Rüsch-Gerdes
laboratories/health authorities, trains more than 80 persons per
tuberculosis with molecular methods such as the Xpert® MTB/
be isolated from pigs. Until now, reports addressing the issue
S, Tefera ZD, Calain P, de la Tour R, Jochims F, Richter E,
year, and cooperates with national (RKI, Robert Koch Institute;
Sierra Leone, Slowenia, Turkmenistan, Uganda, Uzbekistan,
RIF assay. The assay detects M. tuberculosis and rifampicin
whether specific strains are responsible for human infection
Bonnet M. High prevalence of multidrug-resistant tuberculosis,
public health offices; public health offices) and international
resistance in parallel and can be carried out nearly fully
are scarcse. The assumption that environmental strains and not
Swaziland, 2009-2010. Emerg Infect Dis. 2012 Jan;18(1):29-37.
centers/organizations (WHO; MSF, Médecins Sans Frontières;
automated, integrating bacterial lysis, nucleic acid extraction,
a subgroup of highly virulent strains are the causative agents
amplification, and amplicon detection using a disposable plastic
is not proven yet. Prerequesite for the clarification of these
cartridges. The NRL evaluated the assay for extrapulmonary
specimens, for which the method was not yet evaluated in
ICRC, International Committee of Red Cross; USAID, U.S. Agency
Kurbatova EV, Cavanaugh JS, Shah NS, Wright A, Kim H,
for the International Development; FIND; The Foundation for
questions is the establishment of reliable, easy to perform and
Metchock B, Van Deun A, Barrera L, Boulahbal F, Richter E,
Innovative New Diagnostics).
standardizable typing method. Since restriction fragment lentghs
Martín-Casabona N, Arias F, Zemanova I, Drobniewski F,
To improve the quality standard of diagnostic laboratories in
clinical studies and could show that the combined sensitivity
Santos Silva A, Coulter C, Lumb R, Cegielski JP. Rifampicin-
Europe the NRL coordinates a EU project for external quality
and specificity of the Xpert assay were calculated to be 77.3 %
resistant Mycobacterium tuberculosis: susceptibility to isoniazid
control for drug susceptibility testing. Additionally the NRL was
and 98.2 %, respectively.
and other anti-tuberculosis drugs. Int J Tuberc Lung Dis.
appointed member of the „Management Team“ by the European
In recent years the global situation concerning drug resistance has
2012;16(3):355-7.
Centre for Disease Prevention and Control (ECDC) program to
improve TB-diagnostic in Europe. Since 2010 samples for quality
changed dramatically. In order to combat this, the NRL focused
to research on the development of resistance and epidemiology
Siddiqi S, Ahmed A, Asif S, Behera D, Javaid M, Jani J,
control for 31 countries were produced, sent, and the incoming
of TB, combining classical and advanced microbiological as
Jyoti A, Mahatre R, Mahto D, Richter E, Rodrigues C,
data analysed.
well as molecular methods. Drug susceptibility testing (DST) of
Visalakshi P, Rüsch-Gerdes S. Direct drug susceptibility testing
resistant Mycobacterium tuberculosis (Mtb) strains is usually
of Mycobacterium tuberculosis for rapid detection of multidrug
Grant support
based on the analysis of growth in culture media containing
resistance using the Bactec MGIT 960 system: a multicenter
BMG, DZK-SH, ECDC, EU TB PAN-NET, INSTAND, RKI (National
defined drug concentrations (critical concentrations). Different
study. J Clin Microbiol. 2012 Feb;50(2):435-40.
Reference Laboratory and network respiratory infections), WHO
levels of drug resistance are not regarded. The correlation
of M. tuberculosis isolates with different resistance profiles
Blakemore R, Nabeta P, Davidow AL, Vadwai V, Tahirli R,
and resistance levels were investigated by genetic mutation
Munsamy V, Nicol M, Jones M, Persing DH, Hillemann D,
screening, fingerprint methods and semi-quantitative DST the
Ruesch-Gerdes S, Leisegang F, Zamudio C, Rodrigues C,
BACTEC MGIT 960 system monitored by using EpiCenter TBeXiST
Boehme CC, Perkins MD, Alland D. A Multi-Site Assessment of
software. We analysed strains from Germany isolated in 2011
the Quantitative Capabilities of the Xpert(R) MTB/RIF Assay. Am
that were either multidrug resistant (MDR), or monoresistant for
Figure 1: MLST Analysis for M. avium strains show great variability
isoniazid or rifampicin. Most strains resistant to rifampicin had
of patterns
81
J Respir Crit Care Med. 2011, 184: 1076.
Medical Infrastructure • National Reference Center for Mycobacteria
Dr. Sabine Rüsch-Gerdes
Clinical and experimental pathology
transcriptome /
Validation
Human Ex Vivo Model
Prof. Dr. Dr. Ekkehard Vollmer
Bettina Baron-Lühr • Niklas Bauer • Arne Cerny • Gabriele Cornehls • Steffi Fox • Dr. Jürgen Galle
Dr. Dariimaa Ganbat • Ghazaal Rezaie • PD Dr. Karoline Gaede • PD Dr. Torsten Goldmann • Lydia Guse
Iris Jonas • Dipl. Ing. Daniel Kähler • Angela Kelp • Anke KuhfuSS • Dr . Dagmar S. Lang • Maria Lammers
Milena Lemburg • Carmen Licht • Belinda Lie • M. Sc. Sebastian Marwitz • Christine Möller • Frauke
Pedersen • Romina Pritzkow • Heidi Scheibl • Anna Schiller • Carmen Schöne • Dr. Holger Schultz
Dipl. Biol. Sophie Seehase • Dr. habil. Bernard Schmitt • Dr. Florian Stellmacher • Gabriele Tänzer • Jasmin Tiebach • Rolf Warnecke • Kristin Wiczkowski • Monika Wolgast • Leyla Ziga
MISSION
Moreover, we have provided proof that pHp is part of the
internal and external collaborations
Central experimental tool is a functional human tissue culture model for different diseases of the
lung, based on the HOPE-technique. For screening purposes and biomarker identification we apply
transcriptome analyses, which are complemented by high throughput validation tools such as tissue
microarrays. We deliver state-of-the-art diagnostics focusing on diseases of the lung and develop new
markers for personalized medicine.
surfactant system in the human lung. Ongoing studies focus
Internal collaborations: NRZ (S. Rüsch-Gerdes, E. Richter, D.
towards the role of pHp in COPD and Asthma.
Hillemann), Molecular Mycobacteriology (S. Niemann), Microbiology and Infectiology (U. Schaible), Microbial Inflammation
Research (S. Ehlers, N. Reiling), Cellular Allergology (H.Haas),
Clinical and Molecular Allergology (A. Petersen, U. Jappe),
Mucosa Immunology (A. Frey, K. Ramaker), Clinical Infectious
most important findings
MAdL: The antibody was initially raised against a cytoplasmic
Diseases (C. Lange, U. Greinert, B. Karlsdorf), IExp. Pneumolo-
During the past two years we continued the work on two immuno-
fraction of primary human alveolar epithelial cells type II. Im-
gy (H. Fehrenbach, Ch. Vock), Barrier Integrity (I. Lautenschlä-
regulatory molecules, which were discovered in the human lung
munohistochemistry using MAdL and tissue microarrays from
ger), Cellular Pneumology (H. Haas), Medical and Biochemi-
by our group, BAMBI (Bone morphogenetic protein and Activin
HOPE-fixed Lung cancer specimens showed reactivity with
cal Microbiology (H. Brade), Structural Biochemistry (O. Holst),
Membrane-Bound Inhibitor) and pulmonary haptoglobin (pHp).
adenocarcinomas. A protocol for formalin-fixed-tissues was
Immunochemistry (U. Zähringer, C. Alexander), Biophysics (T.
developed and a broad series of stainings was performed
Gutsmann, J. Andrä, K. Brandenburg, B. Lindner), Biochemical
Together with the group of Johannes Gerdes (Borstel) and
comparing MAdL with the established markers. We demons-
Immunology (F. Petersen), Vet. Infection Biology and Immunolo-
Gernot Zissel (University Freiburg) we developed an antibody
trated that an additional diagnostic benefit was achieved
gy (J. Ahmed, U. Seitzer), Cellular Pneumology (C. Stamme), In-
designated MAdL (Marker for Adenocarcinomas of the Lung),
using MAdL in 16 % of the cases; due to the large number of
nate Immunity (H. Heine), Fluorescence Cytometry (T. Scholzen)
which came to the market this year.
patients with NSCLC this would make about 132.000 cases per
External collaborations: Airway research Center North (ARCN)
year worldwide (Figure 1). We conclude that MAdL is a useWe developed a procedure which allows for HOPE-preservati-
ful marker in the diagnostics of NSCLC which is commercially
Figure 2: Immunohistochemical detection of pHp in a HOPE-fixed human
on and paraffin-embedding of BAL-samples.
available since 2012.
lung which has been stimulated ex vivo with dexamethasone. Signals are
visible (red color) in alveolar epithelial cells type II (200x).
in the German Center for Lung research (DZL). University of
Lübeck, Med Clinic III (K. Dalhoff, D. Drömann). University of
Lübeck, Medical Microbiology (J. Rupp). LungenClinic Grosshansdorf (Rabe K, Reck M, H. Watz, Ch. Kugler, F. Pedersen, L.
We have established a facility which provides primary human
selected puBlications
Welker). University of Dohuk, Iraq (M. Abdullah). Tblisi Cancer
Schultz H, Marwitz S, Baron-Lühr B, Zissel G, Kugler C, Rabe
Center, Georgia (A. Mariamidze). Yerevan State Medical Univer-
Since the start of the German Center for Lung Research (DZL)
KF, Zabel P, Vollmer E, Gerdes J, Goldmann T. Generation
sity, Armenia (A. Mkhitarian). University of Freiburg, Pneumology
we are engaged in multiple research projects concerning
and evaluation of a monoclonal antibody designated MAdL as
(J. Müller-Quernheim, G. Zissel, A. Prasse). BMBF Medizinische
COPD, Asthma, and Lung Cancer. Funded by the DZL, we
a new specific marker for adenocarcinomas of the lung. British
Infektionsgenomik: University of Braunschweig, Robert Koch Ins-
have now the possibility to perform transcriptome analyses in-
Journal of Cancer (2011) 105(5), 673 – 681
titute Wernigerode University of Greifswald (M. Steinert, A. Flie-
alveolar epithelial cells type II and other primary cells.
ger, M. Hecker, L. Jänsch).
house with an Agilent-platform which nicely complements the
Abdullah A, Kähler D, Vock C, Kugler Ch, Drömann D, Rupp
methodological spectrum of the Research Center. The Clin. &
Exp. Pathology is responsible for many biobanking activities
(Biomaterialbank Nord) covering diverse projects in the DZL
and others.
Figure 1: Expression patterns of IHC markers in 167 cases of adenocarcinomas of the lung. The diagnostic benefit of MAdL is displayed with “#” and
accounts for 27 cases (16 %)[from Schultz et al., Br J Cancer 2011; 83:6].
83
J, Hauber HP, ReilingN, Dalhoff K, Zabel P, Vollmer E, Gold-
Grant support
mann T. Pulmonary haptoglobin and CD163 are first line immuno-
BMBF: DZL Airway Research Center North: Funding for projects
regulatory elements in the human lung. Respiration (2012) 83(1):6
in the areas of COPD, Lung Cancer, Asthma and Biobanking.
BMBF project on the ex vivo infection of human lung tissues with
We have established a branch in the LungenClinic Grosshans-
pHp: After the initial description of pHp we showed that pHp
Abdullah M, Goldmann T. Pulmonary haptoglobin (pHp) is
Legionella pneumophila (Legioprotect). Roche: Co-funding in a
dorf which serves for both, routine diagnostics and a large
and its receptor CD163 are expressed and regulated by micro-
part of the surfactant system in the human lung. Diagn Pathol
project on the development of a procedure to acquire broncho-
variety of research activities in the DZL and others.
bial antigens, IL-6 and glucocorticoids in the human lung (Figu-
(2012) 7:158
alveolar – lavages and sputum with the HOPE-technique.
re 2). Further, pHp and CD163 are actively secreted from lung
Furthermore, in close cooperation with the Nationales Refe-
tissues upon stimulation and enhance the secretion of soluble
Marwitz S, Abdullah M, Vock C, Fine JS, Visvanathan S, Ga-
renzzentrum für Mykobakterien, we expanded the range of
mediators. Php also induces the release of immunomodula-
ede K, Hauber HP, Zabel P, Goldmann T. HOPE-BAL: Impro-
molecular diagnostics in the lung by implementation of EML4/
tory molecules in lung cells. As a translational approach we
ved Molecular Diagnostics By Application Of A Novel Technique
Alk -rearrangement and B-RAF-mutations, which are decision-
showed that the immunohistochemical detection of pHp can
For Fixation And Paraffin-Embedding. J Histochem Cytochem
making in targeted therapies of lung cancer.
be used to identify adenocarcinomas in routine diagnostics.
(2011) 59 (6):601 – 614
Medical Infrastructure • clinical and
xxxxxxxxxxxxxxxxxxxxx
experimental pathology
HOPE technique
Center for Clinical Studies
Allergy
Johanna Döhling • Andrea Glaewe • Lenka Krabbe
Bronchial Carcinoma
MRCP(UK)
MISSION
for bronchoalveolar lavage. This study resulted in two papers on
The most innovative clinical project is ExplainTB! that aims to
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.
the regional pulmonary immune response in healthy subject that
bridge the language barrier faced by up to 50% of tuberculosis
are latently infected with M. tuberculosis.
patients in Europe (Figure 2). Making use of smartphones, QR-
The allergy focus of the Center for Clinical Studies was
codes and educational videos, a powerful tool for point-of-
addressed by the multicenter BASALIT study investigating
care patient education is under development in co-operation
the associations between soy and birch allergens (principal
with the Deutsches Zentralkomitee zur Bekämpfung der
Development
investigator Uta Jappe; FKZ 01KG0911). The trial is ongoing.
Tuberkulose and TB-NET (producer: Stefan Gieren, Fiction2.0).
One main focus in 2012 was the expansion and consolidation
Two further investigator initiated multicenter trials are also
of our infrastructure. In May 2012, Christian Herzmann was
conducted at the Borstel study site. For COSYCONET, a cohort
selected puBlications
employed as the new head of department. In March 2013,
trial investigating the impact of co-morbidity on COPD, we
Herzmann C, Aagaard CS, Andersen P, Ernst M. Divergent
Johanna Döhling, successfully completed the AGAH study
recruited >20 patients. Unfortunately, we recruited only 1
ESAT-6 epitope recognition in human PBMC and BAL cells.
nurse training. For 2013, an additional part-time study nurse
patient for PROGRESS-CAP aiming to identify predictors
EMBO Conference on Tuberculosis, Institute Pasteur, Paris, Sept
contract was agreed on with Alere Technologies GmbH, Jena.
and risk factors for deterioration in patients with community
11-15, 2012. Poster 287
The collaboration with our librarian Kay Geschuhn lead to
acquired pneumonia (>10 screening failures).
the website for our unit in December (sz.fz-borstel.de). For
Small numbers of patients were enrolled in several non-invasive
Herzmann C, Ernst M, Ehlers S, Stenger S, Maertzdorf J, Sot-
the future, this enables us to establish new scientific links to
studies on the treatment of advanced bronchial carcinoma but
giu G, Lange C. Increased frequencies of pulmonary Treg cells
research groups in Borstel, to regional respiratory physicians
the importance of this disease calls for improvement of our
in latent M. tuberculosis infection. Eur Respir J. 2012 Mar 22.
and to the pharmaceutical industry. As a consequence of the
oncological study performance.
ongoing construction work in the hospital, our unit is spatially
Since May 2012, we intensified patient enrolment into the
Herzmann C et al. Immunological evidence of incipient pulmo-
restricted since mid 2012.
WeanNet registry that captures comprehensive data on
nary tuberculosis. J Infect Dis, Sept 10, 2012.
Most important projects
Figure 1: Fluorescence linked immunosorbent assay used for the detection
of mycobacterial lipoarabinomannan in the urine of tuberculosis patients.
Honouring our traditional research emphasis that was
prolonged weaning from invasive ventilation. Thanks to the
collaboration with Lukas Hundack, Borstel hospital is currently
internal and external collaborations
preparing its accreditation as a certified Weaning Center.
Industry partners: AID – Alere Technologies – Linde
established by our previous head, Christoph Lange, tuberculosis
n=42; NTM infection, n=14; suspected TB with alternative
In December 2012, the sponsor driven PulmonX LIVE study
Therapeutics – Novartis – Pfizer – PulmonX – Siemens
(TB) remained in the focus of our interest. Sponsored by
diagnosis, n=15). The method had a very limited diagnostic
was initiated that serves as a registry for patients treated with
Academic partners: Bernhard-Nocht-Institute – Deutsches
Alere Technologies GmbH, we conducted a pilot study
yield with high false positive rates among patients with
endobronchial valves for emphysematous bullae. Enrolment of
Krebsforschungszentrum – Health authorities Hanover Frankfurt
evaluating the diagnostic use of transrenal mycobacterial
bronchial carcinoma.
patients is pending. A medicinal device by Linde Therapeutics
– Institute for Lung Research – Koch-Metschnikoff-Forum – Max-
DNA fragments (principal investigator Christoph Lange; trial
Supported by a grant from the DFG, the diagnostic value of a
is currently under investigation for the standardised 6-Minutes-
Planck-Institut for Infection Biology – Statens Serum Institut,
identifier NCT01585532). Between February and March 2013
dual IL-2 / IFN-γ fluorescence Elispot was evaluated in various
Walking-Test.
Copenhagen – Tb or not Tb Consortium – TB-NET – University
we recruited about 80 patients for urine and blood sampling.
groups of tuberculosis patients (principal investigator: Barbara
Hospital of Ulm, Institute for Hygiene and Medical Microbiology
Interim analyses are promising.
Kalsdorf). In 2012, 66 participants were enrolled in the trial.
Internal collaborations: Biophysics – Clinical Infectious
A second collaboration on TB urine diagnostics was little
The study is ongoing.
Diseases – Immune cell analytics – Immunobiophysics –
successful. Given the poor sensitivity of ELISAs for the
In December 2012, a further diagnostic pilot study was
Immunochemistry – Microbial interface biology – Biobank
detection of mycobacterial liparabinomannan (LAM) in the
started (principal investigator Christoph Lange; trial identifier
urine, a monocentre clinical trial evaluating the sensitivity
NCT01748357). In collaboration with Siemens, the trial aims to
and specificity of a fluorescence linked immunosorbent
detect a pattern of exhaled volatile organic compounds. Since
assay (FLISA) was conducted (principal investigator Christian
December 2012, about 25 patients have been recruited.
Herzmann; trial identifier NCT01587677). The method that
Our unit also participates in the cohort study of the consortium Tb
was developed at the Deutsches Krebsforschungszentrum,
or not Tb (head: Stefan Ehlers; co-ordinator: Karoline Gaede; both
Heidelberg, allows single-molecule detection of LAM (Figure
Borstel; FKZ 01KI1007D). Close collaboration with the Hamburg
1). In collaboration with Jakko van Ingen, Nimjegen, 116
health authority (Karen Meywald-Walter) and the Bernhard-
Figure 2: Illustration of the ExplainTB! project. Scanning of QR-Codes ena-
patients for 4 study groups were recruited between February
Nocht-Institute (Christian Meyer, Thorsten Thye) facilitated the
bles healthcare providers to supply tuberculosis patients with educational
and October 2012 (confirmed TB, n=45; bronchial carcinoma,
recruitment >50 patients at our study site, including >20 subjects
videos in their own language.
85
Medical Infrastructure • Center for Clinical Studies
Dr. Christian Herzmann,
Tuberculosis
COPD
Weaning
Impressum
Academic degree and professional qualification 2011/2012
total budget 2011
institutional funding ( federal + federal state)
15.722 j
third-party funding
6.684 j
of which DFG
30 %
41 %
2.293 j
34 %
EU project funding
469 j
7 %
industry
785 j
12 %
foundations
268 j
4 %
69 j
1 %
miscellaneous
2012
10
8
Diploma
5
2
Bachelor of Science
1
1
Master of Science
5
3
10
10
1
1
2011
2012
internal
19
12
external
8
25
2011
2012
national
24
19
international
55
60
Dissertation
2.800 j
federal/federal state
2011
Technicians
Animal Keeper
Conferences and Workshops 2011/2012
total budget 2012
institutional funding (federal + federal state)
17.030 j
third-party funding
7.480 j
31 %
Guest scientists 2011/2012
of which DFG
3.050 j
40 %
2.290 j
31 %
EU project funding
930 j
12 %
industry
600 j
8 %
foundations
420 j
6 %
miscellaneous
140 j
2 %
federal
Peer review publications 2011/20120
2011
141
2012
125
Patents and Licenses 2011/2012
Books and book articles 2011/2012
Assignation of patents
1
Stock of patent families
21
2011
5
6
2012
10
Stock of licensing agreements
Income from royalties (T j)
327 (2011)
372 (2012)
87
Facts & Figures
Funding (in thousands)
Impressum
International Networks 2011/2012
Administrative and Technical Services
German Research Foundation ( DFG)
7th EU-Frame Program
Exzellenzcluster ‘Entzündung an Grenzflächen’
GALtrain (Marie Curie EST): Exposure to micro-organisms and
Managing Director
SFB / TR 22, Allergische Immunantwort der Lunge
suppression of allergic diseases: A European research training
Prof. Dr. Stefan Ehlers
SFB 654, Plastizität und Schlaf
scheme
SPP 1313, Biological Responses to Nanoscale Particles
ConFluTech: Capacity building for the control of Avian influenza
Board of Directors
SPP 1394, Mast cells – promotors of health and modulators of
through technology transfer and training [coordination, FZB]
Prof. Dr. Heinz Fehrenbach
disease
Arbo-Zoonet: International network for capacity building for the
Prof. Dr. Ulrich Schaible
SPP 1580 ‘Intracellular compartments as places of pathogen-
control of emerging viral vector borne zoonotic diseases
Prof. Dr. Peter Zabel
host-interactions’
Cure HLH: European initiative to improve knowledge, treatment
GRK 1727, Modulation der Autoimmunität’
and survival of haemophagocytic syndromes in children
Administration
GRK 1743 ‘Gene, Umwelt und Entzündung’
INFLA-CARE: Understanding inflammation-associated tumori-
Jürgen Repp (Head)
genesis for the rational design of novel anti-cancer therapeutic
Klaus Bartels (Technical Building and Construction Management)
Federal Ministry for Education and Research
strategies
Dirk Grünes (Technical Building and Construction Management)
Deutsches Zentrum für Lungenforschung
TB-PAN-NET: Pan-European network for the study and clinical
Sibylle Lenkeit (Financial Management)
Deutsches Zentrum für Infektionsforschung
management of drug resistant tuberculosis
Tanja Schroeter-Ohrendorf (Human Ressources))
CAPNETZ – Ambulant erworbene Pneumonie
POSTICK: Post-graduate training network for capacity building to
Heinz-Jürgen Wefel (Infrastructural Management)
Schwerpunkt Biophotonik - OPTOPROBE
control ticks and tick-borne diseases
Verbundprojekt ‚Suszeptibilität bei Infektionenen: Tuberkulose‘
TheSchistoVac: The targeted development of a new generation
IT
Förderprogramm ‚Ersatzmethoden zum Tierversuch‘
Vaccine for Schistosomiasis
Michael Reinhold (Head)
Innovative Therapieverfahren auf molekularer und zellulärer Ba-
EUMEDNETvsTB: Building a cooperative strategy between Eu-
Arno Gepp
sis mit Schwerpunkt Sepsis
rope and Mediterranean Countries for upgrading tuberculosis
Timo Hammerbacher
Medizinische Infektionsgenomik
research and control
Lars Kagrath
Förderinitiative NanoCare ‚Prädiktion humantoxikologischer Wir-
PiroVac: Improvement of current and development of new vac-
Ivan Montti
kung synthetischer Carbon Black Nanopartikel‘
cines for theileriosis and babesiosis of small ruminants [coordi-
ERA-NET PathoGenoMics
nation: FZB]
Animal Housing
EDENext: Biology and control of vector-borne infections in Europe
Dr. Ilka Monath
PathoNgen -Trace - Next Generation: Genome based high resolution tracing of pathogens.
Research Library and Documentation
Kai Geschuhn (Head)
Others
Christiane Engler
Centre for Nanovaccines, Programkomitteen f. Strategiske Vaekstteknologier, Kopenhagen. ‘Imaging vaccine efficacy’
Works Council
Gates Foundation: Imaging TB
Heiko Käßner (Chairman)
Joint research grant National Research Foundation South Af-
Manuel Hein (Vice Chairman)
rica/DFG “Development and validation of a sensitive LAMP assay for the specific detection of Theileria sp.(sable) in sable
(Hippotragus niger) and roan (Hippotragus equinus) antelope”
Deutsch-Afrikanische Kooperationsprojekte in der Infektiologie
(PAK467) Projekt “Molecular epidemiology network for promotion and support of delivery of live vaccines against Theileria
parva and Theileria annulata infection in Eastern and Northern
Africa” (coordination, FZB)
89
Administrative and Technical Services
National Networks 2011/2012
Organization chart 2013
Board of Curators Chair
Rolf Fischer
Scientific Coordination
Dr. B. Brand
Managing Director
Prof. Dr. S. Ehlers
Scientific Advisory Board Chair
Prof. Dr. K. Pfeffer
Impressum
Herausgeber
Prof. Dr. S. Ehlers
Equal Opportunities Officer
N. Grohmann
Koordination
Dr. B. C. Brand
Bilder
Fotolia
S.4; A. Raths
S.5; PSDESIGN1, Krishncreations
S.6; G. Klein
S.10; the photos
S.18; Irochka
S.19; fotodo
S.20/21; molekuul_be
S.86; Fotolia 2867025
S.87; R. Razvan
S.88; Moodboard
S.89; monropic
FZB
S.7; 15; 16; 22; 23
H. Fehrenbach
S.11
Design
STILPUNKT3 Designbüro
Hamburg
Druck
Partner Werbung & Druck
Pinneberg
Board of Directors/Extended Board
Managing Director: Prof. Dr. S. Ehlers
Program Director: Prof. Dr. U. Schaible, deputy: PD 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
Priority Area Infections
Director: Prof. Dr. U. Schaible
Medical
Infrastructures
Director:
Prof. Dr. P. Zabel
Priority Area Asthma & Allergyy
Director: Prof. Dr. H. Fehrenbach
Works Council
Administration
Head: J. Repp
Central Units
Analytical
Chemistry
Dr. D. Schwudke
Clinical Infectious
Diseases
Prof. C. Lange
Hospital
Prof. P. Zabel
Clin. & Mol.
Allergology
Prof. U. Jappe
Structural
Biochemistry
Prof. O. Holst
Human Ressources
T. SchröterOhrendorf
IT
M. Reinhold
Biophysics
Prof. T. Gutsmann
ImmuncellAnalytics
Dr. M. Ernst
Medical Service
Center
Prof. E. Vollmer
Mucosal Immunology & Diagnostics
PD Dr. A. Frey
Biochemical
Immunology
Prof. F. Petersen
Financial
Management
S. Lenkeit
Sci. Information
Services & Library
K. Geschuhn, MA
Center for
Clinical Studies
Dr. C. Herzmann
Cellular
Allergology
PD Dr. H. Haas
Innate Immunity
Prof. H. Heine
Infrastructural
Management
J. Wefel
Animal Facility
Dr. I. Monath
Immunobiophysics Microbial Interface
Biology
PD Dr. A . Schromm
PD Dr. Reiling
Models of
Inflammation
Prof. G. Graßl
Infection
Immunology
Dr. C. Hölscher
Clin. & Exp.
Pathology
Prof. E. Vollmer
Cellular
Pneumology
Prof. C. Stamme
Cellular
Microbiology
Prof. U. Schaible
Biobank
PD Dr. K. Gaede
Vet.-Infection
Biology &
-Immunology
Prof. J. Ahmed
Molecular
Mycobacteriology
Mycobacteriology
(NRC)
PD Dr. S. Niemann Dr. S. Rüsch-Gerdes
Immunobiology
Experimental
Prof. Bulfone-Paus
Pneumology
Prof. H. Fehrenbach
Invertebrate
Models
Dr. C. Wagner
Molecular
Immunology
K. Lee, PhD
Mouse Models
in Asthma
Dr. M. Wegmann
Fluorescence
Cytometry
Dr. T. Scholzen
Techn. Building
& Construction
Management
D. Grünes
Hospital
Management
n.n.
BBRS
Dr. S. Paetzold
In-Firm Training
Dr. S. Zähringer
Quality
Development
U. Schroer
Leibniz-Zentrum
für Medizin und
Biowissenschaften
Parkallee 1-40
23845 Borstel
Tel.: +49 (0)4537.188 0
fzb@fz-borstel.de
www.fz-borstel.de
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