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Das schönste, was wir entDecken können, ist Das Geheimnisvolle

EinbettenHerunterladen
Forschungsbericht
2009/2010
Das Schönste,
was wir entdecken
können, ist das
Geheimnisvolle
Albert Einstein
Forschungsbericht
2009/2010
,etsnö hcS saD
n e kcedtne riw saw
sad tsi ,nennö k
ellovsinmieheG
n i e ts n i E t r e b l A
Leibniz-Zentrum
für Medizin und
Biowissenschaften
Impressum
Vorwort
Seite 6
Tuberculosis Center Borstel (TBCB)
Seite 8
Borstel Biomedical Research School
Seite 10
OPTOPROBE – Visionen möglich machen
Seite 12
Peptide – Wunderwaffe gegen die Sepsis?
Seite 14
Highlights
2009
Seite 16
Wissenschaftlicher Beirat/Scientific Advisory Board
2010
Seite 18
Prof. Dr. Werner Seeger
Research Reports
Direktor der Klinik für Innere Medizin II,
Justus von Liebig Universität Gießen
Dept. Molecular Infection Biology
(Vorsitzender)
Cellular Microbiology
Medical and Biochemical Microbiology
Structural Biochemistry
Immunochemistry
Biophysics
Immunobiophysics
Molecular Inflammation Medicine
Infection Immunology
Microbial Interface Biology
Models of Inflammation
Prof. Dr. Barbara Wollenberg
Direktorin der Klinik für Hals-, Nasen- und Ohrenheilkunde,
Universität zu Lübeck
(stv. Vorsitzende)
Prof. Dr. Christoph Dehio
Dept. Immunology and Cell Biology
20
22
24
26
28
30
32
34
36
38
Immunobiology
Molecular Immunology
Biochemical Immunology
Biological Chemistry
Veterinary Infection Biology
and Immunology
Immune Cell-Analytics
Innate Immunity
Fluorescence Cytometry
Infection Biology, Biozentrum, Universität Basel
Prof. Dr. Thomas Hünig
Institutsvorstand des Institutes für Virologie und Immunologie,
Bayerische Julius-Maximilians-Universität, Würzburg
Prof. Dr. Erika Jensen-Jarolim
Head of the Department of Pathophysiology and Allergy Research,
Center for Pathophysiology, Infectiology & Immunology
Medical University of Vienna
Prof. Dr. Thisbe Lindhorst
Direktorin des Institutes für Organische Chemie,
Christian-Albrechts-Universität zu Kiel
Prof. Dr. Klaus Dieter Pfeffer
Direktor des Institutes für Medizinische Mikrobiologie
und Krankenhaushygiene, Universität Düsseldorf
40
42
44
46
48
50
52
54
Dept. Pneumology
Inflammation and Regeneration
Cellular Pneumology
Pathophysiology of Inflammation
Barrier-Integrity
National Reference Center
for Mycobacteria
Molecular Mycobacteriology
Clinical and Experimental Pathology
Cellular Allergology
Clinical and Molecular Allergology
Mucosa Immunology
Clinical Infectious Diseases
56
58
60
62
64
66
68
70
72
74
76
Facts & Figures
Funding
Seite 78
Patents
Seite 78
Academic degree and professional qualification
Seite 79
Conferences and workshops
Seite 79
Guest scientists
Seite 79
Peer review publications
Seite 79
Books and book articles
Seite 79
National networks
Seite 80
International networks
Seite 80
Administrative and technical services
Seite 81
Imprint
Seite 82
Organization Chart
Seite 83
xxxxxxxxxxxxxxxxxxxxx
Inhalt
5
MAGAZIN
Impressum
Der vorliegende Forschungsbericht informiert in kurzer, kompak-
es den Bogen von der physikalisch-chemischen und molekular-
ter Form und in neuer Aufmachung über die wissenschaftlichen
biologischen Grundlagenforschung über den Einsatz komplexer
Aktivitäten des Forschungszentrums Borstel in den Jahren 2009
Modellsysteme und epidemiologischer Studien hin zu translati-
und 2010. Der Magazinteil berichtet über wichtige Forschungs-
onaler klinischer Forschung und Patientenversorgung. Die lang-
ergebnisse, Verbundaktivitäten und neue Entwicklungen an
fristigen Ziele des FZB – Verbesserung bestehender und die
unserem Zentrum. Exemplarisch ausgewählte Projekte der For-
Entwicklung neuer Methoden zur Erkennung, Vermeidung und
schergruppen und Serviceeinheiten zeigen sowohl die Kernkom-
Behandlung von Lungenerkrankungen – basieren auf dem inter-
petenz auf als auch die erbrachte Leistung.
disziplinären Forschungsansatz des Zentrums, der multidisziplinären Gruppenstruktur und der breit angelegten methodischen
Auf diese Weise möchten wir nicht nur versierten Kollegen, son-
Expertise.
dern auch einer breiteren Öffentlichkeit unsere wissenschaftli-
I m m u n o lo gy a n d C e l l B i o lo gy
chen Fragestellungen und Ziele näher bringen, aber auch die
Stolz sind wir auf die hervorragende Arbeit aller unserer Mit-
Fortschritte aufzeigen, die wir in den beiden vergangenen Jah-
arbeiter, ein herzlicher Dank gilt allen Freunden und Förderern
ren bei der Lösung gesellschaftlich relevanter medizinischer
aus Wissenschaft, Politik und Wirtschaft. Mit Ihrer Unterstützung
Probleme erzielt haben.
blicken wir optimistisch in die Zukunft!
Das FZB erforscht entzündliche Erkrankungen infektiösen und
Peter Zabel
nicht-infektiösen Ursprungs vor allem der Lunge. Dabei schlägt
Ulrich Schaible
PREFACE
The present research report informs about the scientific ac-
lung. In doing so, the center forges a bridge from physico-
tivities of the Research Center Borstel (RCB) in the years
chemical and molecular-biological basic research, via the
2009 and 2010 in a short, compact form and in a new lay-
use of complex model systems and epidemiological studies
out. The magazine section reports on important research re-
to translational medicine and patients’ care. The long-term
sults, cooperation activities, and new developments at our
goals, i. e. the improvement of existing methods and the
Center. Exemplarily chosen projects of the research groups
development of new and innovative ones for diagnosis,
and service oriented units demonstrate both the core com-
prophylaxis, and treatment of lung diseases, are based on
petence and the achieved accomplishments.
the interdisciplinary approach of the Center, the multidisciplinary group structure, and the broad spectrum of method-
By this means we would on the one hand like to give an
ological expertise.
understanding of our research approach and objectives
P n eu m o lo gy
not only to experienced colleagues but also to a broader
We are proud of the excellent work of all our colleagues,
public and on the other hand to show the progress we have
and we are deeply grateful to all friends and supporters
made during the last two years in contributing to solving
from science, politics, and industry. With your support we
medical problems relevant to society.
are looking optimistically to the future!
The RCB performs research on inflammatory diseases of
Peter Zabel
infectious and non-infectious origin, with a focus on the
Ulrich Schaible
7
xxxxxxxxxxxxxxxxxxxxx
Vorwort • Preface
M o l ecu l a r I n f ect i o n B i o lo gy
VORWORT
reportage
dizinischen Klinik und dem Nationalen Referenzzentrum, um die
TB-Forschung in Deutschland in internationalen Forschungsver-
Diagnostik, Epidemiologie, Infektionsüberwachung und Thera-
bünden zur Eindämmung der TB-Epidemie repräsentiert.
pieberatung nachhaltig zu verbessern. Damit würden Ergebnis-
TBCB als Kern-Bestandteil des DTZ soll den wirksamen Schutz
se der Grundlagenforschung zeitnaher in die klinische Anwen-
der Bevölkerung vor der Tuberkulose befördern, insbesondere
dung gebracht. Eine solche strategische Weiterentwicklung am
durch die Entwicklung und den Aufbau
FZB wäre gesundheitspolitisch hochrelevant für die Prävention
•epidemiologischer Frühwarnsysteme besonders für mehr-
und Behandlung der Tuberkulose.
fach-resistente Erreger,
•schnellerer und automatisierter Verfahren in der Tuberkulo-
Tuberculosis Center Borstel (TBCB)
sediagnostik mit molekularbiologischen Techniken,
•einer international ausgerichteten Zentralstelle zur mole-
der TB-Forschung arbeiten zusammen geschlossen. Diese neue,
kularen und klinischen Überwachung der Ausbreitung re-
programmatisch als Querschnittsbereich ausgerichtete Struktu-
sistenter Tuberkuloseerreger mit zentraler und gesicherter
Wissenschaftler gestalten Zukunft!
rierung der infektiologischen Expertise am FZB soll den Aus-
Patientendatenbank und Biobank,
von Prof. Dr. Stefan Ehlers
um folgende Zielsetzungen zu erreichen:
stoffe und immunmodulatorischer Substanzen unter Nut-
•Identifikation von Virulenz- und Persistenzmechanismen
zung der internationalen Netzwerke,
tausch zwischen Grundlagen- und klinischer Forschung fördern,
des Tuberkuloseerregers und ihrer molekularen GrundlaTuberkulose-Forschung in Borstel – ist das nicht schon ein alter Hut? In der Tat ist Borstel bereits 1947 als Tuberkulose-Forschungsinstitut gegründet worden. In den fünfziger Jahren wurden z. B. im Herrenhaus die •von Weiterbildungsmaßnahmen von medizinischem Fachpersonal and Kliniken und Gesundheitsämtern über Aus-
•Aufklärung der Entwicklung und Ausbreitung von Antibioti-
breitung, Diagnose und Therapie von Mykobakterieninfekti-
ersten Antibiotika, die gegen TB-Bakterien wirksam waren, im Reagenzglas und im Tiermodell getestet. Später •Identifikation von Biomarkern, die bei latent infizierten Per-
kam eine renommierte Lungen-Fachklinik hinzu – ein bisschen Sanatoriums-Charakter à la Thomas Manns Zau-
sonen die Entwicklung einer aktiven Tuberkulose besser
vorhersagen als aktuelle diagnostische Verfahren,
berberg hat die Borsteler Einöde ja immer noch ...
•von klinischen Studien zur Wirksamkeitstestung neuer Wirk-
gen, um Zielstrukturen für neue Wirkstoffe zu definieren,
ka-Resistenzen für globale Überwachungsstrategien,
•Durchführung von Klinischen Studien mit dem Ziel die Dau-
onen (Tuberkulose und nichttuberkulöse Mykobakteriosen)
auch unter Gesichtspunkten der Gesundheitsökonomie,
•eines Konsiliarsystems zur Absprache mit nationalen und
internationalen Kollegen für die Behandlung von Tuberkulosepatienten mit komplizierten Krankheitsverläufen
•einer international angelegten Beratungsstelle für Ärzte,
Inzwischen haben sich jedoch sowohl die Tuberkulose in ihrer
grundlagenorientierten Struktur-, Molekular-, Zell- und Immun-
er der präventiven Tuberkulosetherapie zu verkürzen und
Erscheinungsform, als auch das FZB in seiner Forschungskompe-
biologie über die molekulare Pathogenese- und präklinische
wirksam auch Erkrankungen durch multiresistente Tuberku-
Kliniken und Patienten zur Behandlung resistenter Tuberku-
tenz dramatisch verändert. Weltweit erkranken etwa 9 Millionen
Therapieforschung bis hin zu klinischen Studien, Diagnostik, Pa-
losebakterien zu verhindern,
losen über Videokonferenzen.
Menschen neu an der Lungentuberkulose – etwa 2 Millionen
tientenversorgung und Leitlinienerstellung. Zu den zum Teil in
•Charakterisierung schützender und pathologischer Immu-
Es dürfte klar geworden sein: dies ist kein alter Wein in neuen
sterben daran. Hierzu trägt die Koinfektion mit dem HI-Virus
Deutschland einzigartigen Infrastrukturen, die das FZB für die
nantworten zur Entwicklung immunmodulatorischer Strate-
Schläuchen! Man könnte eher sagen, dass eine solch umfassen-
vor allem in Afrika und Südasien erheblich bei. Relativ neu ist
TB-Forschung vorhält, gehören:
gien und Wirkstoffe, um die „richtige“ Immunantwort z. B.
de strategische Neuausrichtung der TB-Forschung in Borstel ein
das Auftreten von multiresistenten (MDR) und extrem resistenten
•gentechnische Anlagen der Sicherheitsstufe 3 für umfangrei-
(XDR) Tuberkulose-Erregern vor allem in Asien und Osteuropa:
che analytische Arbeiten am Tuberkuloseerreger, gentech-
Die Wissenschaftler im TBCB haben sich zum Ziel gesetzt, den
se Vorhaben der Erweiterung des S3-Laborbereichs zur Entwicklung und Testung neuer Antiinfektiva und Immunmodulatoren in
im Rahmen einer Impfung zu erzielen.
ganz neues Fass aufmacht. In der Tat bedarf das TBCB für die-
erstere sind gegen die beiden besten TB-Medikamente resis-
nisch veränderten Varianten und infizierten Tieren,
Handlungsstrang in Form eines „bottom-up“ Prozesses zu or-
tent (Isoniazid und Rifampicin), letztere sind gegen die meisten
•das kliniknahe Aerosolinfektionsmodell in der Maus,
chestrieren, in dem sie sich Zielvorgaben und Kooperationspro-
kliniknahen Modellen sowie erweiterter Sicherheitsmaßnahmen
antituberkulotisch wirksamen Medikamente unempfindlich. Dies
•innovative Bildgebungsverfahren zur Darstellung der Infekti-
jekte erarbeiten und daraus die Projektfinanzierung begründen.
im Nationalen Referenzzentrum zur Arbeit mit und Lagerung von
führt zu extrem langen Behandlungszeiten und deutlich höheren
MDR und XDR Tuberkuloseerregern. Eine Epidemiologie- und
onsdynamik in komplexen Modellsystemen,
Sterblichkeiten. In einigen Ländern der ehemaligen Sowjetunion
•eine wachsende Plattform für globale Lipidanalysen (Lipidom),
Biostatistik-Gruppe zur Unterstützung der Planung, Durchfüh-
liegt die Rate von MDR-Tuberkulose bei Ersterkrankungen be-
•ein hochauflösendes NMR zur Analyse kleinster Proben-
rung und Auswertung nationaler und internationaler Studien-
reits bei über 15 %! Hierbei handelt es sich u. a. um Länder wie
Estland und Moldawien, die bereits den Euro eingeführt haben
bzw. in naher Zukunft eine EU-Mitgliedschaft beantragen wer-
vorhaben muss aufgebaut und ein international ausgerichtetes
mengen aus Mykobakterien,
Schulungszentrums für Tuberkulosebelange muss eingerichtet
•innovative Sequenzier- und Transkriptomtechniken zur Erre-
werden. Schließlich müssen Anstrengungen intensiviert werden,
gertypisierung,
den. Eine schnellere Diagnostik und Resistenztestung ebenso
•die weltweit größte Mykobakterien-Stammsammlung,
die ein internationales Netz von Labors und Feldstationen zur
wie die Neuentwicklung von Therapeutika, die die Behandlung
•das Nationale und Supranationale Referenzzentrum für TB
TB-Forschung in Osteuropa und Afrika entstehen lassen, die mit
der TB effizient beschleunigen, sind dringend erforderlich. Ein
neuer Wettlauf mit der Zeit gegen die Macht der, inzwischen
multiresistenten, weißen Pest hat begonnen!
Und das FZB? Es hat sich in den letzten 15 Jahren zu einem international anerkannten Kompetenzzentrum der Tuberkuloseforschung entwickelt, das dieses Schwerpunktthema molekular-epidemiologisch, infektionsbiologisch und klinisch breit abbildet.
(NRZ, SRZ),
Dies spiegelt die Entwicklung zu einem von den Wissenschaft-
dem FZB durch Kooperationsverträge verbunden sind.
•nationale und internationale Verbünde (TBornotTB, TBNET,
lern im diskursiven Abstimmungsprozess selbst definierten „Pro-
TB-PAN-NET) mit nachgewiesenem Rekrutierungspotenzial
grammbudget“ wider, das Forschungs- und Strukturdefizite ana-
Das FZB hat sich positioniert: Tuberkuloseforschung ist ein trag-
für klinische Studien,
lysiert und Förder-Prioritäten setzt.
fähiger Schwerpunkt der Mission des Zentrums geworden. Mit
Das TBCB ist Kernstück der FZB-Beteiligung am Deutschen
der Einbindung in das DZIF und dem erklärten Ziel, ein Deut-
Zentrum für Infektionsforschung. Hier koordiniert das FZB den
sches Zentrum für Tuberkuloseforschung zu gründen, erfolgt
•ein klinisches Studienzentrum, das für Klinische Infektiologie
zertifiziert ist,
•Vertretungen in nationalen und internationalen Gremien
(RKI, DGI, DZIF, WHO, ERS).
9
Forschungsbereich „Tuberkulose“ mit Kooperationspartnern in
der konsequente nächste Schritt, die Diagnostik, Therapie und
Hamburg, Köln/Bonn, Hannover/Braunschweig, Tübingen und
Prävention der Tuberkulose nicht nur in Deutschland signifikant
Die Forschung am FZB an dieser hochrelevanten bakteriellen
Was noch fehlt, ist eine am Patienten orientierte engere Zu-
München. Ziel ist es, ein international erkennbares Deutsches
zum Wohle der Patienten zu verbessern. Für die TB-Forschung in
Lungeninfektion des Menschen schlägt den Bogen von der
sammenarbeit der Borsteler Grundlagenforschung mit der Me-
Tuberkulose-Zentrum (DTZ) aufzubauen, das die translationale
Borstel gilt mehr denn je: Think globally, act locally!
TB Center
Im neu gegründeten Tuberculosis Center Borstel haben sich daher insgesamt 12 Laborgruppen des FZB, die überwiegend in
Interview
Die Planung der eigenen wissenschaftlichen Karriere Dorothee: Ich arbeite in der Gruppe von Prof. Grassl in Bors-
Summa summarum klingt es, als wenn die Graduate School
ist eine groSSe Aufgabe. Es stellen sich viele Fragen –
tel und bin auch Teilnehmer der BBRS, denn das Forschungs-
ein voller Erfolg ist. Gibt es auch Verbesserungsvorschläge?
etwa nach dem richtigen Konzept und den verschiede-
zentrum Borstel bietet ein Fortbildungsprogramm und ein Be-
Daniel: Die Graduate School ist für alle Doktoranden des FZB
nen Möglichkeiten. Das FZB bietet seinen Doktoranden*
treuungsangebot in Form von regelmäßigen, zielorientierten
konzipiert. Für mich persönlich ist es ein wenig schade, dass
seit 2010 im Rahmen der BBRS nicht nur ein thematisch Mentorengesprächen. Dadurch bekommt meine Promotionszeit
die Tumorbiologe nicht mehr zu den Schwerpunkten der FZB
fokussiertes Forschungsprogramm an, sondern auch Struktur, was ich als sehr vorteilhaft empfinde.
Mission gehört und daher auch nicht im Programm der BBRS
Dorothee, entschuldige, aber fühlt Ihr Euch nicht ständig
diesen Themenbereich fällt, bewegt sich somit leider mehr im
Im Interview berichten Daniel Kähler und Dorothee
kontrolliert?
Randbereich der Themen der BBRS. Für die meisten Teilnehmer
Schultz von ihren bisherigen Eindrücken und Erfah-
Nein. Struktur erfordert einfach gewisse Absprachen und es
trifft dies allerdings nicht zu, schließlich arbeite ich ja in der
rungen. Daniel promoviert in der Gruppe ‚Klinische handelt sich ja hier um eine beiderseitige Verpflichtung, die von
Pathologie, die mehr als nur eine forschende Arbeitsgruppe
und Experimentelle Pathologie‘ (Prof. Vollmer), ist im den Dozenten genauso wahrgenommen wird, z. B durch die
ist – Stichwort Diagnostik – und das Thema „Tumor“ daher zur
dritten Jahr seiner Promotion und gehört somit zu Qualität ihrer Seminare oder der Aufarbeitung einer Thematik.
täglichen Arbeit dazugehört.
den Quereinsteigern in die Graduate School, wäh-
Mir gefällt es, dass so viele unterschiedliche Mitarbeiter des
rend Dorothee seit Beginn ihrer Promotion in der FZB an dem Programm beteiligt sind: Zum Beispiel Laborgrup-
Als zukünftige akademische Führungskräfte, vermisst Ihr
Gruppe ‚Entzündungsmodelle‘ (Prof. Grassl) dabei ist.
penleiter oder die Bibliothekarin. Das schafft Abwechslung! Alle
Themen wie Personalführung, Führungsstrategien etc.?
Dozenten sind mit Herzblut dabei, von den Grundlagen ausge-
Daniel: Also ich persönlich finde diesen ganzen Themenbe-
hend die aktuellen und relevanten Aspekte eines Themas dar-
reich spannend und auch sehr wichtig. Ich habe an der HAW
Mit welchen Erwartungen seid Ihr an die Grduate School
zulegen und mit den wesentlichen Kernaussagen zu verknüpfen.
(Hamburger Hochschule für Angewandte Wissenschaften) mal
herangegangen?
Daniel: Mir gefällt auch der seminaristische Charakter der
einen exzellenten Kurs zur Personalführung belegt: hoch interes-
Dorothee: Die Graduate School ist eine sinnvolle Einrichtung.
Veranstaltungen, die überschaubare Gruppe – wir sind aktuell
sant, besonders auch der Teil zu Motivationsstrategien. Sicher-
Sie hat ein durchdachtes, klares Konzept. Gleich zu Beginn
etwa 15 Teilnehmer – und es bleibt immer ausreichend Zeit für
lich ist das für uns irgendwann von großer Bedeutung.
berücksichtig wird. Meine Doktorarbeit, die aber genau in
meiner Promotion bekam ich so einen Überblick über die Forschungsschwerpunkte des FZB und somit einen Blick über den
thematischen Tellerrand meiner Promotionsarbeit.
Kanntest Du das FZB schon vorher?
Ich habe in Hamburg studiert aber über Bekannte schon vom
FZB gehört.
dical
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i
B
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B ors te
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h
Sc
h
c
r
Resea
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a
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i
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BBRS: er
Daniel, wie ist das bei Dir, Du arbeitest ja schon länger in
ttina Brand
e
B
r.
D
e
rt
w füh
as Intervie
Borstel?
Genau, ich bin jetzt im dritten Jahr meiner Promotion und habe
schon am Mentorenprogramm, dem Vorläufer der Graduate
D
Fragen und Diskussionen.
Dorothee: Aber dafür sorgt die BBRS jetzt schon, dass wir hier
Dorothee: So wird uns ermöglicht die Gruppenleiter in einer
nicht als introvertierte Einzelgänger über den Campus laufen.
anderen, entspannten Atmosphäre kennen zu lernen. Das baut
Barrieren ab, erleichtert den Kontakt eben auch in wissenschaft-
Apropos Karriere – habt Ihr schon Pläne für die Zeit mit
lichen Fragen, die das eigene Projekt betreffen.
Doktor-Titel?
Daniel: Nun, es ist nicht von der Hand zu weisen, dass die
Wie hoch ist denn der zeitliche Aufwand für die Graduate School?
Industrie viele attraktive Angebote auch in der Forschung bietet,
Daniel: Das umfangreiche Programm und die intensive Betreu-
und sich teilweise finanziell unabhängiger darstellt. Die akade-
ung erfordern schon einen recht hohen Einsatz von beiden Seiten.
mischen Einrichtungen haben es daher etwas schwerer in Punk-
Dorothee: Es sind Zwischenberichte zu schreiben, wir organisie-
to Stellenangebote. Oftmals scheint man da um die Einwerbung
ren eine eigene Tagung mit internationalen Sprechern. Das ist
von Drittmitteln nicht herum zu kommen. Aber ich möchte hier
sicher zeitintensiv, aber es macht auch Spaß.
nicht falsch verstanden werden, es geht nicht nur ums Geld. Ich
würde z. B. gern in der Forschung bleiben, die akademische
School, teilgenommen. Ich habe dann beschlossen, das volle Programm der Graduate School ebenfalls mitzumachen. Inzwischen
Kommt da nicht die Laborarbeit zu kurz? Ich denke da ins-
Laufbahn bietet da jedoch nicht viel Spielraum: es gilt hopp
oder topp, d.h. in vollem Tempo den Post-Doc und die Habilitati-
bin ich Doktorandensprecher, Dorothee ist übrigens meine Stell-
besondere an die Endphase der Promotion, wenn es an
vertreterin, ich empfinde das Programm, das durch ein Zertifikat
das Zusammenschreiben geht.
on durchziehen. Da herrscht ein enormer, ganz spezieller Druck
bestätigt wird, als echte Bereicherung meiner Ausbildung.
Dorothee: Ich kann nur jedem empfehlen das Graduate Pro-
auch aufgrund der vielen Drittmittel-finanzierten Verträge. Und
gramm an den Beginn der Promotionszeit zu legen. Gerade in
da nicht jeder von uns eine Professur bekommen kann oder
Zertifikat? Was ist das genau?
der Anfangszeit profitiert man am meisten von den Seminaren.
möchte, ist eine langfristige Perspektive ohne die stramme Lauf-
Daniel: Nun, im Prinzip ist das nur eine schriftliche Bestätigung,
Wir hatten kürzlich unser erstes eigenes Retreat. Das hat mit
bahn unwahrscheinlich. Dann muss man überlegen, wie man
dass ich an einem von der Leibniz-Gemeinschaft entworfenen,
sehr gut gefallen.
Familie und Privatleben mit dem Beruf vernünftig in Einklang
strukturierten Doktorandenprogramm in vollem Umfang teilge-
Daniel: Mir auch, besonders, weil man Gelegenheit hatte, mit
bringen kann. Ich könnte mir z. B. eine Art Probezeit an einer
nommen habe. Art und Anzahl der Veranstaltungen sind in ei-
allen Kollegen mit und ohne Wissenschaft Zeit zu verbringen.
Universität oder einem Forschungszentrum vorstellen, der dann
nem ‚course record book‘, das auch unseren Vertrag mit dem
Man lernt sich kennen, kann sich austauschen und geht nicht nur
aber langfristige Verträge folgen. Mit 2-Jahres-Verträgen auf
FZB enthält, festgehalten. Die jeweiligen Veranstaltungen wer-
einfach nett grüßend auf dem Campus aneinander vorbei. Die
Dauer ist eine langfristige Planung sehr schwierig!
den von den betreuenden Dozenten abgezeichnet. Im Prinzip
Kommunikation unter den Doktoranden hat sich mit der Gradu-
Dorothee: Zum jetzigen Zeitpunkt halte ich mir alle Möglich-
verpflichtet sich jeder Teilnehmer*, das vollständige Programm
ate School spürbar verbessert.
keiten offen, die sich für mich nicht nur auf den Bereich Wissen-
zu absolvieren, es steht aber jedem frei, in welchem Zeitraum
Dorothee: Das Retreat hatte den Charakter einer kleinen Kon-
schaft und Forschung beschränken.
er/sie das bewerkstelligt. Hat man mal keine Zeit, kann die Ver-
ferenz. Wir haben in der Vorbereitung unsere Abstracts einge-
anstaltung in einer anderen Einheit nachgeholt werden. D. h. 15
reicht und dann – wie immer auf englisch – unsere Vorträge
ganztägige Veranstaltungen in drei Jahren sollte man schaffen.
gehalten. Es gab wirklich gute Diskussionen.
*Alle Amts- und Funktionsbezeichnungen gelten in gleicher Weise für weibliche und
männliche Personen
11
Biomedical Research School
ein strukturiertes Qualifizierungskonzept.
reportage
Visionen möglich machen: von der Idee zum Produkt
Konsortium aus Wissenschaft und Wirtschaft entwickelt neue optische
Sonden für die Tumordiagnostik von Dr. Barbara Frey, Dr. Bettina Brand und PD Dr. Andreas Frey
Licht für die Gesundheit: das ist die Mission der Biophotonik – „bios“
für Leben und „phos“ für Licht. Licht misst schnell, empfindlich und
berührungslos – ein ideales Werkzeug für die Aufklärung von biologischen Prozessen und Krankheitsursachen, eine Chance, Veränderungen frühzeitig zu erkennen und gezielte Therapien zu entwickeln.
www. biophotonik.org
Das hochgesteckte Ziel des OPTOPROBE Konsortiums ist die
für die nächste Evolutionsrunde vorschlägt, welche eine große
Entwicklung neuartiger Kontrastmittel und ihre Einbindung in
Wahrscheinlichkeit besitzen, die gewünschten Eigenschaften in
ein Endoskop-basiertes diagnostisches System, das neben der
die nächste Generation weiterzutragen. Der „Züchtungsvorgang“
Krebsfrüherkennung auch die simultane, laserinduzierte Zerstö-
erlaubt neben der gezielten Kreuzung auch Punktmutationen
rung des Tumors während des endoskopischen Eingriffs ermög-
durch zufällig gesteuerten Aminosäureaustausch und entspricht
lichen soll. „Wir sind sehr glücklich darüber, dass wir unsere er-
damit dem biologischen Evolutionsprinzip vom Überleben der Fit-
folgreichen Vorarbeiten aus dem Vorläufer-Verbund PLOMS, der
testen. Durch die Anwendung dieses gerichteten Evolutionsverfah-
sich mit der Darmkrebs-Früherkennung beschäftigt hat, in das
rens gelingt es im Verlauf weniger Generationsrunden, optimierte
erweiterte OPTOPROBE Konsortium überführen konnten. Dieses
Tumorsonden durch Synthese und Testung einer überschaubaren
interdisziplinäre Vorhaben ist nicht nur technologisch wegwei-
Anzahl von Peptiden zu entwickeln. Die optimierten Peptide wer-
send, sondern wird dem Patienten von großem Nutzen sein,“ so
den anschließend mit molekularen Schaltern versehen, die z.B.
Dr. Andreas Frey, Koordinator des Konsortiums am Forschungs-
die Aktivierung dieser Sonden erst nach Bindung an die Zielstruk-
zentrum Borstel. Die neue Technologie soll die Krebsfrühdiag-
tur ermöglichen. Durch die Kopplung an Fluoreszenzfarbstoffe
nostik erheblich verbessern und damit die Chancen erhöhen,
wird die Detektion der so entstandenen Kontrastmittel durch ein
Patienten bereits im frühen Stadium einer Krebserkrankung
angepasstes Mehrkanalfluoreszenzendoskop ermöglicht.
erfolgreich zu behandeln. Derzeit lassen sich Tumore endoskopisch erst erkennen, wenn sie eine gewisse Größe erreicht ha-
Das langfristige Ziel von OPTOPROBE ist die Verfügbarkeit von
ben. Das bedeutet aber auch, dass sich die Tumorzellen bereits
peptidbasierten Kontrastmitteln mit einem darauf abgestimmten
stark vermehrt und vielleicht schon weiter ausgebreitet haben.
Endoskop für die Detektion von kleinsten (< 1 mm Durchmesser)
Wenn man bedenkt, dass heute die Fünf-Jahres-Überlebensrate
Darmtumoren, Kehlkopf- und Lungentumoren sowie OP-begleiten-
bei Darmkrebs bei 60 Prozent liegt, kann man davon ausgehen,
de Schnelldiagnostik. Die weiteren Planungen sehen die Kombina-
dass durch eine rechtzeitige Erkennung des Tumors die Überle-
tion des Systems mit einer thermischen Therapie (laserinduzierte
benschance auf über 90 Prozent ansteigt.
Zerstörung des Tumors) vor, wodurch dem Patienten eine Operation mit größerem Risiko erspart wird. Und was sagt das Zeitma-
Das Verbundvorhaben beinhaltet die translationale Entwicklung
nagement? Dr. Frey rechnet damit, dass, wenn alles gut läuft, in
eines neuen diagnostischen Verfahrens. Voraussetzung ist das sy-
etwa fünf Jahren erste Geräte und in etwa fünf bis acht Jahren die
nergistische Zusammenwirken der individuellen Partnerexpertisen:
entsprechenden Peptidsonden auf dem Markt sein können.
von der Identifizierung einer Zielstruktur, über die Bereitstellung
der benötigten Reagenzien und Diagnostika bis hin zur Geräteent-
PD Dr. Andreas Frey
wicklung sowie der Erprobung, Anwendung und Markteinführung.
Förderung des Projekts
Das Bundesministerium für Bildung und Forschung fördert das
Projekt mit 2,6 Mio. Euro im Rahmen des Forschungsschwer-
Das Peptid Puzzle
punkts Biophotonik. Die Wirtschaftspartner investieren zusätzlich
Kernstück des Projekts ist die Entwicklung neuartiger, „intelligen-
1,75 Mio. Euro in das Projekt.
ter“ Kontrastmittel, die geeignet sind, das endoskopische Diagnoseverfahren zu einer bisher unerreichbaren Empfindlichkeit
Partner
voranzutreiben. Als Basis für derartige Kontrastmittel sollen Pep-
ATTO-TEC GmbH, Siegen, Produktion von Fluoreszenzfarbstoffen
tidsonden dienen, da sich Peptide (Aminosäureketten) in ihrer
für die biomedizinische Diagnostik, www.atto-tec.com
biologischen Wirkung durch exzellente Spezifität und hohe Effek-
GeSiM (Gesellschaft für Silizium Mikrosysteme mbH), Großerk-
tivität auszeichnen. Das Problem besteht darin, bei all den mögli-
mannsdorf/Dresden, Produktion von Nanoliter-Dosiertechnik für
chen Peptidvariationen - ein Peptid einer Kettenlänge von nur 10
die Herstellung von Bio-Chips, www.gesim.de
Bausteinen, das aus den 20 natürlichen Aminosäuren zusammen-
LMTB (Laser- und Medizin-Technologie GmbH Berlin), Entwick-
gesetzt ist, erlaubt bereits 10 Billionen Kombinationsmöglichkei-
lung optischer Geräte und Verfahren für die Medizin und Bio-
ten – geeignete Kandidaten zu identifizieren: die sprichwörtliche
technologie, www.lmtb.de
Suche nach der Nadel im Heuhaufen. Daher wurde speziell für
R-BIO (R-Biopharm AG), Darmstadt, Lebens- und Futtermittelana-
diese Fragestellung ein molekulares Evolutionsverfahren entwi-
lytik und klinische Diagnostik, www.r-biopharm.com
ckelt, bei dem Peptide mit maßgeschneiderten Eigenschaften in
KST (Karl Storz GmbH & Co KG), Tuttlingen, Weltmarktführer im
einer hochautomatisierten Syntheseplattform „gezüchtet“ werden
Bereich starre Endoskope, www.karlstorz.de
können. Hierzu wird zunächst eine Startpopulation an Peptiden
auf definierte Eigenschaften wie z. B. Bindung an Tumorzellen,
Kontakt
Stabilität des Peptids im menschlichen Körper oder Zellgängig-
PD Dr. Andreas Frey, afrey@fz-borstel.de, Tel. 04537.188562
keit analysiert. Die Testergebnisse werden in ein Computerpro-
Bildmaterial © FZB/Frey
gamm eingespeist, das die Ursprungspeptide in Abhängigkeit
Mehr zum Thema
von ihrer Qualität untereinander kreuzt und neue Peptidmoleküle
Biophotonik – Licht für die Gesundheit www.biophotonik.org
13
optoprobe
OPTOPROBE –
reportage
Wunderwaffe gegen
die Sepsis?
Ein Bilderbuchbeispiel
der translationalen
Forschung.
von Dr. Bettina Brand
Peptide – die neue Wunderwaffe?
soll schließlich die therapeutische Wirksamkeit des neuen Medi-
„Dass ausgerechnet Medikamente wie Antibiotika, die zur The-
kamentes an ca. 200 bis 10.000 Patienten signifikant belegt wer-
rapie der Sepsis eingesetzt werden, die Freisetzung des LPS
den (pivotal study). Nach einem erfolgreichen Abschluss einer
fördern anstatt es zu binden, ist die eigentliche Krux an der Ge-
Klinischen Phase III Studie kann die Marktzulassung für Europa
schichte“ sagt Klaus Brandenburg. Bereits in den 90er Jahren
bei der „European Medicines Agency“ (EMEA) oder für die USA
fanden Wissenschaftler Sequenzen in Proteinen, die über diese
bei der „Food and Drug Administration“ (FDA) beantragt wer-
wertvolle Bindungsfunktion verfügen. Infolge der Charakterisie-
den. Die Marktchancen sind gewaltig: allein in den USA schätzt
rung der Binderegion des Lactoferrins gelang die Synthese von
man die Ausgaben zur Behandlung der Sepsis auf 17 Milliarden
LPS-bindenden Peptiden, die allerdings nur in hohen Konzentra-
Dollar. Aufgrund der zunehmenden Antibiotikaresistenzen könnte
tionen und begleitet von einer Reihe von Nebenwirkungen wirk-
die Bedeutung der Peptidtherapie noch weit darüber hinausge-
sam waren. Das Ziel von Klaus Brandenburg: Peptide effektiver
hen. So scheint es auch für das grosse Problem der multiresisten-
zu machen, um die Endotoxin-Flut bei einer Sepsis zu neutralisie-
ten Keime – Stichwort MRSA = Methicillin-resistente Staphylococ-
ren und somit die Entzündungsreaktion einzudämmen. „Auf Basis
cus aureus - eine Lösung zu geben, da einige der entwickelten
unserer Forschungsergebnisse mussten wir ein multi-tasking Pep-
Peptide sehr gute Abtötungseigenschaften aufweisen. „Das heißt
tid entwickeln: es musste positiv geladen sein, um das negativ
nicht, dass die klassischen Antibiotika vor der Ablösung stehen.
geladene LPS binden zu können, es musste, wie das LPS, am-
In der direkten Bekämpfung bakterieller Erreger sind Antibiotika
phiphil sein d.h. sowohl wasser- als auch fettlöslich, um wirkungs-
immer noch effektiver“, so Klaus Brandenburg. Bleibt die Frage,
voll zu reagieren.“ Mit diesen Ansätzen gelang es, synthetische
wann mit der endgültigen Verfügbarkeit für die Patienten zur
Peptide zu entwickeln, die bereits in einem Verhältnis von 10:1
rechnen ist. Klaus Brandenburg ist da optimistisch, “Wenn wir
(Peptid:LPS) wirksam sind. Dass es sich um vielseitige Peptide
die klinischen Phasen erfolgreich abschliessen sollten, könnte
handelt, zeigten Untersuchungen mit Gram-positiven Bakterien,
die Therapie spätestens 2014 zur Verfügung stehen.“
die in ihrer Zellwand anders aufgebaut sind als Gram-negative.
Bei ihnen lösen die Murein-Lipoproteine Entzündungsreaktionen
aus. „Wir haben bereits einige Lipoproteine getestet und gesehen, dass es zu einer effektiven Bindung kommt. Somit sind wir
optimistisch, dass auch Gram-positive Sepsis mit diesen Peptiden behandelbar sein wird.“ Die Peptide können darüber hinaus
auch bei Vireninfektionen wirksam sein, die das Immunsystem
Das PeptidTeam (v. li.):
K. Brandenburg,
Y. Kaconis,
T. Gutsmann,
I. Kowalski.
initial schwächen und erst durch eine bakterielle Superinfektion
zu einer schweren Erkrankung führen. Eine direkte Schädigung
der Viren konnte bisher nicht gezeigt werden, jedoch werden
die humanen Andockstellen der Viren blockiert und dadurch die
Virusvermehrung erheblich unterdrückt.
Förderung des Projekts
Wie kommt es zu einer Sepsis?
Man spricht von einer Sepsis, wenn die Erreger oder von ihnen
From bench to bedside.
Bundesministerium für Bildung und Forschung. Verbundprojekt ‚In-
produzierte Gifte bei einer Infektionskrankheit den ursprüngli-
Nachdem die Wirksamkeit der Peptide sowohl in vitro als auch
novative Therapieverfahren auf molekularer und zellulärer Basis‘
chen Entzündungsherd verlassen und sich über das Blut im
in vivo nachgewiesen ist, laufen zur Zeit Zytotoxizitätstests, die
www.bmbf.de
Organismus ausbreiten. Dadurch entwickelt sich im gesamten
eine mögliche zellgiftige Wirkung der Peptide auf menschliche
Körper eine heftige Immunreaktion, die außer Kontrolle gerät.
Immunzellen nachweisen sowie weitere vorklinische Studien,
Projektentwicklung
Folge kann ein septischer Schock sein, bei dem das Kreis-
z. B. zur chemischen und physikalischen Stabilität der Peptide.
in Zusammenarbeit mit Ascenion www.ascenion.de
laufsystem, die Blutgerinnung und die Organe versagen. Am
In 2011/2012 sollen die klinischen Phasen beginnen: In Phase
häufigsten lösen Bakterien eine Sepsis aus, seltener auch Pilze
I wird die Verträglichkeit eines Wirkstoffes zunächst an einer
Kontakt
oder Parasiten, meistens als Folge einer Lungenentzündung, Er-
kleinen Gruppe von gesunden Studienteilnehmern überprüft.
Prof. Dr. Klaus Brandenburg
krankungen im Bauchraum, Infektionen der Harnwege und der
Ermittelt werden die Wirkungen und Nebenwirkungen des Me-
kbrandenburg@fz-borstel.de, Tel. 04537.188235
Haut. Schlüsselfiguren sind die Lipopolysaccharide(LPS), auch
dikamentes sowie die geeignete Dosierung. In Phase II wird
Prof. Dr. Thomas Gutsmann
Endotoxine genannt. Sie sind Zerfallsprodukte der Zellmemb-
die Wirksamkeit des Therapiekonzepts für eine spezifische oder
tgutsmann@fz-borstel.de, Tel. 04537.188291
ran Gram-negativer Bakterien wie Salmonellen, Escherichia coli
mehrere Indikationen an 50- 200 erkrankten Patienten überprüft
oder Legionellen. Freigesetzt werden Endotoxine bei der Zelltei-
(proof-of-concept), der optimale Dosierungsbereich ermittelt so-
Mehr zum Thema
lung der Bakterien, bei Angriffen auf das Immunsystem oder bei
wie Verträglichkeiten und Nebenwirkungen untersucht. Zur Zeit
Leibniz Journal zum Thema Gesundheit
Antibiotikabehandlungen. Der menschliche Körper leitet gegen
wird im Gespräch mit der deutschen Zulassungsbehörde, dem
www.leibniz-gemeinschaft.de/journal
die als Feind erkannten Moleküle eine Phalanx von Abwehr-
Bundesinstitut für Arzneimittel und Medizinprodukte (BfArM), ge-
biotechnologie.de – die Informationsplattform
reaktionen ein, die aufgrund der hohen LPS-Konzentration bei
prüft, ob die Phasen I und II nicht zusammengelegt werden kön-
www.biotechnologie.de
einer Sepsis verheerend sind.
nen, um das Zulassungsverfahren zu beschleunigen. In Phase III
Deutsche Sepsisgesellschaft www.sepsis-gesellschaft.de
15
SEPSIS
Peptide –
In Deutschland gibt es einen
weitgehend unbekannten Killer:
Nach einer Studie des Kompetenznetzes Sepsis (SepNet) sterben
hierzulande jeden Tag durchschnittlich 162 Menschen an
einer Sepsis (Blutvergiftung)
– den Folgen einer auSSer Kontrolle geratenen Infektion durch
Bakterien oder andere Mikroorganismen. Der Biophysiker Klaus
Brandenburg und sein Team sind
diesem Killer seit Jahren auf den
Fersen und haben nun erfolgreich Wirkstoffe entwickelt, um
ihn dingfest zu machen. Dazu
hat er ein BMBF-gefördertes
Konsortium mit Thomas Gutsmann (FZB) und zwei weiteren, in
der Sepsisforschung erfahrene
Gruppen aus Hannover (Mathias
Hornef) und Aachen (Tobias
Schürholz) gegründet.
Highlights
Stapellauf der Cluster Lecture, ein prestigeträchtiges Diskussionsforum des Exzellenzclusters
‚Inflammation at Interfaces‘. International renommierte Forscher sprechen über ihre wegweisenden
The Collaborative Research Centers 654 ‘Plasticity and sleep’ and TR22 ‘Pulmonary allergies’
were positively
Arbeiten und diskutieren mit Studenten, Kollegen und Gästen im Herrenhaus Borstel.
Start der ersten
re-evaluated and will be funded for another period of four years.
Graduate School
The award for the best doctoral thesis of the year (Modellsysteme für Infektionskrankheiten) des Leibniz Center Infection.
University of Lübeck offered a
professorship (W3)
(Promotionspreis des Kreises Segeberg) was given to Claudia Plinke.
in biophysics to Thomas Gutsmann.
Beste Diplomarbeit 2009: Corinna Bang.
OPTOPROBE - Joint venture of nonprofit-scientific institutions
auf die W3 Professur für Biophysik
der Universität zu Lübeck.
The Young
Wolf-Meinhard Becker und Jochen Artur Ulmer. Beides Forscher der ersten Stunde,
die Borstel mitgeprägt und vorangebracht haben.
Willkommen geheißen wurde Uta Jappe, ihres Zeichen Allergologin
und Nachfolgerin von Wolf-Meinhard Becker.
(BMBF), to develop new optical probes for diagnostics in cancer.
17
Researcher Award in Tuberculosis was granted
to Claudia Jafari by the European Respiratory Society.
HIGHLIGHTS
Abschied und BegrüSSunG: zwei Borsteler Urgesteine werden mit spannenden und ungewöhnlichen Abschiedssymposien in den Ruhestand entlassen:
and business companies, funded by the Federal Ministry of Education and Science Ruf an Thomas Gutsmann
The Ranking of the Deutsche Forschungsgemeinschaft (DFG) provides information about the participation
of German research institutions in the funding programmes of the DFG and other national and international research funding institutions. It also reports in differentiated form on the thematic priorities of non-university research institutions with regard
OPTOPROBE - BMBF-gefördertes Konsortium
Der Preis für die beste Doktorarbeit des Jahres
- Promotionspreis des Kreises Segeberg - geht an Claudia Plinke.
aus Wissenschaft und Wirtschaft entwickelt neue
to their third-party funded research activities.
In 2009 the
optische Sonden für die Tumordiagnostik.
Sonderforschungsbereiche (SFB) 654 „Plastizität und Schlaf“ und
werden als
FZB was ranked third in medical sciences.
exzellent beurteilt und für weitere vier Jahre finanziell von der DFG gefördert.
enabled the center to strengthen structure biology Mega Hz NMR. Claudia Jafari erhält den
Young Researcher Award
Das Konjunkturpaket 2009 macht es möglich:
Neues 700
“International Conference on Intracellular Niches of Microbes” in
Mega Hz NMR malaria, but also have, as symbionts, important functions in diverse ecosystems. This meeting brought
verstärkt die Strukturbiologie.
Launch of the Cluster Lecture, a prestigious new forum to discuss ground-breaking developments created by the Excellence Cluster ‚Inflammation
Zentrum für Pneumologie und Thoraxchirurgie.
der Wissenschaften in Hamburg geht an Stefan Ehlers.
formation of the University
research and discuss with students, colleagues and guests in the Manor House Borstel.
Lung Center North comprising the FZB,
International renommierte Wissenschaftler treffen sich am
the University Medical Center Schleswig-Holstein in
FZB, um über
Corinna Bang received the award for the
Kiel and Lübeck as well as the Hospital Großhansdorf, intrazelluläre Überlebensstrategien und Nischen von Mikrorganismen zu diskutieren Center for Pneumology and Thorax Surgery.
best diploma thesis 2009
from the Association for the Study of Inflammatory and Allergic Processes.
(Förderung: Böhringer Ingelheim Stiftung, Organisation: U. Schaible, B. Brand und A. Haas, Bonn).
The first ‘Hamburg Science Award’ of the
‘Akademie der Wissenschaften’ in Hamburg was granted to Stefan Ehlers.
Leistungsranking der DFG: Unter den 20 erfolgreichsten außeruniversitären Einrichtungen belegt das
FZB
at Interfaces‘. Every month, internationally renowned researchers present their seminal work in inflammation Der erste Hamburger Wissenschaftspreis der Akademie
Borstel.
together renowned specialists for individual intracellular microbes who presented their latest research.
den Partnern: FZB; UK-SH Kiel und Lübeck sowie Krankenhaus Großhansdorf,
by the Leibniz Center Infection.
Intracellular microbes are important causes of human diseases such as tuberculosis, typhoid fever and in Tuberculosis der European Respiratory Society.
Gründung des Universitären Lungenzentrums Nord mit
The Böhringer Ingelheim Foudation supported generously the
Graduate School
“Model Systems for Infectious Diseases” organized and managed by purchasing a new 700
SFB-Transregio 22 „Allergische Immunantwort der Lunge“
Start of the first
National economic stimulus package 2009 has Farewell and welcome: Wolf-Meinhard Becker and Jochen A. Ulmer
Rang 13. Im Fachgebiet Medizin konnte sich das Zentrum sogar auf Platz drei – hinter dem Max-Delbrück-Centrum
- veterans in immunology and allergology – went into retirement not without their own and very special farewell-symposia.
f. Molekulare Medizin, Berlin, und dem Deutschen Krebsforschungszentrum in Heidelberg positionieren.
A warm welcome was given to Uta Jappe, allergist and successor of Wolf-Meinhard Becker.
2010
Highlights
Kimberly Kallsen received the award for
Diplomarbeit des jahres: Kimberly Kallsen.
Ariane Langfeldt
the best diploma thesis
Platz 2 erreicht
des bundesweiten
from the Association for the Study of Inflammatory and Allergic Processes.
second!
FZB has received the certificate ‘berufundfamilie’ for
measures
FZB erhält Zertifikat berufundfamilie für die Maßnahmen zur
Gleichstellung von Frauen und Männern
Leibniz-Training Award:
Ariane Langfeldt came in
Leibniz-Ausbildungspreises.
First national
19
to foster gender equality and family-friendly working environment.
Nano particles are frequently used in our daily life. How these molecules interact
Nano-Partikel werden im Alltag vielfältig eingesetzt. Wie die Teilchen aber auf
with body after being inhaled is largely unknown. The BMBF funded network
die Zellen des menschlichen Körpers wirken, wenn sie eingeatmet wurden, ist nicht im Detail geklärt.
“Nanaocare/Carbon Black”, coordinated by the FZB, is determined to define if the hazard
Die BMBF-geförderte Forschungsverbund „Prädiktion humantoxikologischer Wirkung
potential is depended on precisely defined materials properties.
synthetischer Carbon Black Nanopartikel“, der vom FZB koordiniert wird,
(Contact: H. Fehrenbach)
hat es sich zur Aufgabe gemacht, zu klären, inwieweit das Gefährdungspotenzial
der Carbon Black Nanopartikel von klar definierten Materialeigenschaften abhängt.
(Kontakt: H. Fehrenbach)
The Borstel
Die Borstel
biomedical graduate School
biomedical Research School
started with an interdisciplinary training program.
startet ihr interdisziplinäres Ausbildungsprogramm.
Tödliche Krankheiten wie die Theileriose und Babesiose dezimieren
regelmäßig Rinderherden in Afrika und Asien und bedrohen die Existenz ganzer Familien. Das
FZB
Fatal cattle diseases such as theileriosis and babesiosis decimate
herds in Africa and Asia on a regular basis thus threatening the existence of entire families. The
FZB
coordinates two international consortia, funded by the DFG and EU, to develop new vaccines. koordiniert internationale DFG- und EU-geförderte Forschungskonsortien zur Entwicklung von Impfstoffen.
(Contact: U. Seitzer and J. Ahmed)
(Kontakt: U. Seitzer und J. Ahmed)
Preliminaries to concepts for
Vorentscheidung zu neuen
National Health Research Centers. In tight
deutschen Zentren der Gesundheitsforschung:
conjunction with regional Partners FZB
FZB zweimal vertreten. Gemeinsam mit den
could convince the international advisory board
regionalen Standortpartnern konnte das Zentrum eine
with its performance and innovative concept to
The Volkswagen Foundation has promoted the internationally noticed
internationale Gutachtergruppe von der
Volkswagenstiftung fördert international viel beachteten
shape the German Center for Leistungsfähigkeit und der Konzeptionsstärke
conference on tuberculosis research:
Kongress zur Tuberkuloseforschung:
Infectious Diseases as well as the German Center
bei der Gestaltung deutscher Gesundheitszentren
‘Frontiers in TB Research – The Granuloma:
‚Frontiers in TB Research – The Granuloma:
for Lung Research.
überzeugen und gehört in der Infektions- und
where Pathology meets Microbiology’, June 23-26, 2010 in Borstel.
where Pathology meets Microbiology’, 23.-26. Juni 2010 in Borstel.
Lungenforschung zu den Besten.
promotionspreis
des Kreises Segeberg geht an Susanne Homolka.
(Org.: S. Ehlers, U. Schaible, B. Brand, U. Schroer).
(Org.: S. Ehlers, U. Schaible, B. Brand, U. Schroer).
The award
(Promotionspreis des Kreises Segeberg)
for the best doctoral thesis was given to Susanne Homolka.
HIGHLIGHTS
am Arbeitsplatz und ein familiengerechtes Förderprogramm.
Cellular Microbiology
phagosome
innate immunity
Kristine Hagens • Nadine Harmel • Jacqueline Mahlo • Dagmar Meyer • Carlotta Ober-Blöbaum • Natalja
Redinger • Dr. Karsten Stamer • Michael Wöhlke • Dr. Yeojun Yun
Tuberculosis
host-pathogen interaction
MISSION
on in mycobacteria under replicating and non-replicating condi-
Figure 1:
To analyze host-pathogen interactions in tuberculosis on the molecular, cellular and animal model level with a strong focus on bacterial virulence properties and innate host responses within the micro-environment of the infected lung.
tions using optimized far-red reporters. PLoS One 5:e9823.
Interleukin
18 deficient
Andreu N, Zelmer A, Fletcher T, Elkington PT, Ward TH, Ri-
mice are more
poll J, Parish T, Bancroft GJ, Schaible U, Robertson BD, Wi-
susceptible to
most important findings
immunity. Interestingly, a meta analysis revealed that vaccine
les. S. 2010. Optimisation of bioluminescent reporters for use
tubercle bacilli
The causative bacterial agent of tuberculosis (TB), Mycobac-
efficacies are only faintly affected by micro-nutrient deficiencies
with mycobacteria. PLoS ONE 5(5): e10777
as indicated
terium tuberculosis, is a facultative intracellular pathogen of
(Savy et al., 2009). For further experimental studies on the role
resting macrophages. In order to secure survival and growth
of iron in TB, we developed novel sensors for free iron in biolo-
Fakih S, Podinovskaia M, Kong X, Collins HL, Schaible UE,
numbers of
in macrophages, mycobacteria alter their intracellular fate by
gical samples in close collaboration with Robert Hider at Kings
Hider RC. 2008 Targeting the lysosome: fluorescent iron(III)
red-stained
blocking phagosome maturation. We found that the mycobac-
College London (Fakih et al., 2008).
chelators to selectively monitor endosomal /lysosomal labile
acid-fast bacilli
iron pools. J Med Chem. 14; 51(15): 4539-52.
in the infected
terial cell wall lipid trehalose dimycolate is functionally involved
by increased
in decelerating phagosome maturation (Axelrod et al., 2008).
The world-wide TB crisis and the rise of M. tuberculosis strains
In contrast to resting macrophages, interferon-gamma (IFN-γ)
resistant to more than one anti-TB drugs calls for an accelera-
Savy M, Edmond K, Fine PE, Hall A, Hennig BJ, Moore SE,
Schneider et al.,
activated cells are able to restrict mycobacterial growth by
ted search for novel compounds and more effective prophylac-
Mulholland K, Schaible U, Prentice AM. 2009. Landscape
2010).
several bactericidal mechanisms including nitric oxide (NO)
tics as the current BCG vaccine. Through the Bill-and-Melinda-
analysis of interactions between nutrition and vaccine respon-
production. Macrophage activation also promotes phagosome
Gates funded „Imaging TB“ consortium lead by UES, sensitive
ses in children. J Nutr. 139(11): 2154S-218S.
maturation. Interestingly, we found that NO chemically alters
fluorescent and bioluminescent reporter strains of the TB agent
lungs (see
internal and external collaborations
the cell wall lipids and inactivates its virulence property to de-
were established (Carroll et al., 2010; Andreu et al., 2010). The-
celerate phagosome maturation. This finding revealed that a
se reporters are combined with in vivo imaging devices, the
Lipidomics of intracellular mycobacteria and lysosomal mem-
mycobacterial virulence factor is specifically neutralized by a
commercial IVIS system as well as a Fluorescent Molecular
brane degradation defects: Buko Lindner
host defense effector mechanism. We recently also found that
Tomography (FMT) set up custom made through collaboration
Antimicrobial peptides in tuberculosis: Thomas Gutsmann
NO kills intracellular mycobacteria in an indirect manner by
with laser physicists from Crete (FORTH). This innovative ap-
Mycobacterial glycolipids in phagosome maturation: Otto Holst,
driving apoptotic cell death of infected and activated macro-
proach allows direct imaging of experimental TB in living mice
Imaging tuberculosis: Tanya Parish, QMUL; Brian Robertson,
phages (Herbst et al., 2011, under revision). These data further
infected with those reporter strains in order to monitor infection,
ICL; Gregory Bancroft, LSHTM; Jorge Ripoll, FORTH.
confirm IFN-γ as an essential cytokine in protective immunity
i.e. mycobacterial expansion or elimination, in real time and
Center for Nanovaccines: Peter Andersen, Staten Serum Institu-
against TB. IFN-γ is generated secreted by natural killer, CD4
will be used to assess novel anti-TB compounds as well as
tet, Copenhagen Denmark.
T-helper 1 and CD8 T lymphocytes upon instruction by IL-12
vaccine candidates.
and IL-18. Both cytokines are induced by Toll-like-receptor (TLR)
Iron in infection: Robert Hider, Helen Collins, Kings College London
Lung microbiome: Kenneth Bruce, Kings College London; John
signals. Mice deficient for the TLR-adaptor, MyD88, are highly
selected puBlications
Baines, MPI Evolution Biology, Plön; Pamela Ronald, UC Davis.
susceptible to TB (Hoelscher et al., 2008). We revisited the role
Axelrod, S., H. Oschkinat, J. Enders, B. Schlegel, V. Brink-
Pathogen-Host Interactomes (Infection pathogenomics): Thomas
of IL-18 during experimental TB due to shared usage of MyD88
mann, S.H. Kaufmann, A. Haas, and U.E. Schaible. 2008.
Rudel, Würzburg; Hubert Hilbi, Reinhard Haas, Max-von-Petten-
by signaling cascades for TLR and receptors for IL-1 and IL-18.
Delay of phagosome maturation by a mycobacterial lipid is
kofer Institute Munich, Michael Hensel, Osnabrück; Michael Hec-
Surprisingly, mice deficient for IL-18 and MyD88 succumbed to
reversed by nitric oxide. Cell Microbiol 10:1530-1545.
ker, Greifswald; Thomas Meyer, MPI Infection Biology.
TB much quicker than wild type or TLR-2/-4 deficient mice indicating that lack of IL-18 contributes to the high susceptibility
Schneider, B.E., D. Korbel, K. Hagens, M. Koch, B. Rau-
Grant support
to M. tuberculosis of MyD88 KO mice. In the absence of IL-18,
pach, J. Enders, S.H. Kaufmann, H.W. Mittrucker, and U.E.
BMBF Pathogen-Host Interactomes (“Infection pathogenomics“)
protective T-helper 1 responses were decreased favoring my-
Schaible. 2010 A role for IL-18 in protective immunity against
DFG (“Role of lysosomal phospholipases in infection“)
cobacterial growth and neutrophil-driven lung pathology most
Mycobacterium tuberculosis. Eur J Immunol 40:396-405.
Danish Research Council (“Center of Nano-vaccines“)
likely the cause of premature death of IL-18 knock out mice
Bill-and-Melinda-Gates-Foundation („Imaging TB“)
(Schneider et al., 2010). Recently, we shifted our research focus
Carroll, P., L.J. Schreuder, J. Muwanguzi-Karugaba, S. Wi-
Wellcome Trust (“M. tuberculosis lipids in phagosome matura-
towards the micro-environment of the infected lung. Micronutri-
les, B.D. Robertson, J. Ripoll, T.H. Ward, G.J. Bancroft, U.E.
tion“)
ents such as iron can affect both, pathogen growth as well as
Schaible, T. Parish. 2010 Sensitive detection of gene expressi-
Medical Research Council (“Role of IL-18 in tuberculosis“)
21
Dept. Molecular Infection biology • Cellular Microbiology
Prof. Dr. Ulrich E. Schaible
MEDICAL AND BIOCHEMICAL MICROBIOLOGY
Lipopolysaccharide
Antibody
Ute Agge • Lena Heinbockel • PD Dr. Sven Müller-Loennies • Cristine Schneider
Veronika Susott
Carbohydrate Recognition
Lectin
Structure
Prof. Dr. Helmut Brade
MISSION
immunizations, binding studies and structural analyses. Dr. Colin
selected puBlications
R. MacKenzie from the Institute for Biological Sciences at the
Gerstenbruch S, Brooks C, Kosma P, Brade L, MacKenzie
National Research Council of Canada, ON, Canada, contributes
C, Evans S, Brade H, Müller-Loennies S. Analysis of
with his expertise in biosensor based binding studies (surface
cross-reactive
antibodies
plasmon resonance, Biacore). Upon the initiative of Prof. Dr.
Chlamydophila psittaci.
Thomas Braulke from the Department of Paediatrics at the
against
and
specific
lipopolysaccharide
anti-carbohydrate
from
GLYCOBIOLOGY 2010 Apr;20(4):461-472.
University Clinic Hamburg Eppendorf, Hamburg, Germany we
have isolated a Man6P-specific antibody and in collaboration
most important findings
of carbohydrate antigens. With the exception of CDR H3, the
Brooks CL, Müller-Loennies S, Borisova SN, Brade L, Kosma
established protocols for translation into the clinic. Our work
We have successfully used phage display of single-chain
antibodies are derived from the same set of germline gene
P, Hirama T, MacKenzie CR, Brade H, Evans SV. Antibodies
has received interest from Dr. Gibbes Johnson, Laboratory of
fragments of antibody variable domains (scFv) to isolate two
segments as previously reported structures of S25-2 and S45-
raised against chlamydial lipopolysaccharide antigens reveal
Chemistry, Division of Therapeutic Proteins, Center for Drug
recombinant antibody fragments that recognize with high
18. Despite this similarity, the mAb differ in specificity and the
convergence in germline gene usage and differential epitope
Evaluation and Research, US Food and Drug Administration,
affinity a branched Kdo oligosaccharide which is present only in
mechanism by which they recognize their cognate antigen. S54-
recognition. BIOCHEMISTRY 2010 Jan;49(3):570-581.
Bethesda, MD, USA, with whom we have embarked on a project
Chlamydophila psittaci but not in other pathogenic Chlamydia.
10 uses an unrelated CDR H3 to bind its antigen in a fashion
Sequence determination, binding data, and X-ray structural
analogous to S45-18; however, S73-2 recognizes the same
Brooks C, Blackler R, Sixta G, Kosma P, Müller-Loennies S,
analysis revealed the basis for the improved discrimination
antigen as S45-18 and S54-10 in a wholly unrelated manner.
Brade L, Hirama T, MacKenzie C, Brade H, Evans S. The
between
to use this novel antibody in the preparation of therapeutic
acid
Together, these antibody-antigen structures provide snapshots
role of CDR H3 in antibody recognition of a synthetic analog
(Kdo) ligands and indicated that the alteration of a stacking
into how the immune system uses the same set of inherited
of a lipopolysaccharide antigen. GLYCOBIOLOGY 2010
interaction from a phenylalanine residue in the center of the
germline gene segments to generate multiple possible
Feb;20(2):138-147.
combining site to a tyrosine residue facing away from the
specificities that allow for differential recognition of epitopes
center favours recognition of branched 2.4[2.8]2.4-linked Kdo
and how unrelated CDR H3 sequences can result in convergent
Müller-Loennies S, Galliciotti G, Kollmann K, Glatzel M,
residues. Immunofluorescence tests of infected cell monolayers
binding of clinically relevant bacterial antigens. The analysis
Braulke T. A Novel Single-Chain Antibody Fragment for Detection
using these antibodies showed specific staining of C. psittaci
of the mAb S25-39 structure revealed four different distinct
of Mannose 6-Phosphate-Containing Proteins. Application in
elementary bodies. In order to explore the structural basis for
conformations for the CDR H3 supporting the proposition that
Mucolipidosis Type II Patients and Mice. AMERICAN JOURNAL
adaptability in near germline monoclonal antibodies (mAb),
to minimize entropic penalties associated with immobilization
OF PATHOLOGY 2010 Jul;177(1):240-7.
we have examined the specificity of the promiscuous mAb
of the loops upon binding, some CDRs adapt to antigens by
similar
3-deoxy-a-D-manno-oct-2-ulosonic
A
S67-27 to both naturally derived carbohydrate antigens and
adopting a limited number of ordered conformations rather
Blackler, R.J., Müller-Loennies, S., Brooks, C.L., Brade, L.,
a variety of synthetic nonnatural antigens based on the LPS
than being fully disordered.
Kosma, P., Brade, H. and Evans, S.V. A common NH53K
component Kdo. One such analog, a Kdo(2→8)-7-O-Methyl-
lysosomal enzymes.
mutation in antibodies raised against Chlamydial LPS
Kdo disaccharide, was found to bind to the antibody with at
We have generated a novel scFv antibody fragment against
glycoconjugates significantly increases avidity, Manuscript
least 30-fold higher affinity than any other antigen tested. The
mannose 6-phosphate (Man6P) to allow the specific, rapid,
submitted
structure of S67-27 in complex with this analog and three other
and convenient detection of Man6P-containing glycoproteins.
naturally occurring Kdo antigens revealed that the enhanced
Newly synthesized soluble lysosomal hydrolases require
internal and external collaborations
affinity of the mAb for the synthetic analog was accomplished
Man6P residues on their oligosaccharides for their transport
Strategic alliances have been established with Prof. Dr. Steven
by the strategic positioning of CDR H3 away from a conserved
to lysosomes. Absent or reduced formation of Man6P residues
Evans from the Institute for Biochemistry & Microbiology at
Kdo binding pocket that allowed the formation of new antibody-
leads to the lysosomal storage disorders mucolipidosis type
the University of Victoria, BC in Canada. Prof. Evans is an
antigen contacts. Furthermore, the comparison of this structure
II (ML II) and ML III. The isolated high-affinity Man6P-specific
internationally recognized expert on X-ray crystal structure
with the structures of related mAb revealed how the position
scFv allows the rapid diagnosis of these disorders by simple
analysis of carbohydrate binding proteins. Further support of
B
and structure of CDR H3 influence the specificity or promiscuity
immunoprecipitation and western-blotting. Application of the
our work comes from Prof. Dr. Paul Kosma of the Institute for
of near-germline carbohydrate-recognizing antibodies by
antibody fragment in immunohistochemistry of human brain
Organic Chemistry at the University for Agricultural Sciences in
Figure 1: Shown in (A) is a superposition of the binding site of germli-
altering the architecture of the combining site. The structures
slices from nonaffected patients showed strong neuronal
Vienna, Austria. Prof. Kosma is an expert in chemical synthesis
ne mAb S25-2 (yellow and green) and S25-39 (orange and blue) and in
of two mAb raised against chlamydial LPS, S54-10 and S73-
immunoreactivity, which was not observed in cortical sections
of carbohydrates, has synthesized oligosaccharides which
(B) of liganded and unliganded S25-39 CDR loops (white and yellow),
2, have been determined to high resolution in the presence
of a patient with ML II.
could not be obtained by isolation from natural sources for
with the unliganded CDR loops of S25-2 (orange and grey).
23
Dept. Molecular Infection biology • MEDICAL & BIOCHEM. MICROBIOLOGY
Our research focuses on the analysis of carbohydrate recognition by proteins of the adaptive and innate immune system. We perform structural and functional analyses of antibodies and C-type lectins binding to lipopolysaccharide (LPS) antigens located at the surface of Gram-negative bacteria. The ultimate goal is the identification of high affinity ligands to reveal the underlying molecular mechanism of binding specificity versus cross-reactivity at atomic resolution and to use this information for diagnostic and therapeutic purposes.
Structural Biochemistry
Lipopolysaccharide
glycolipids
polysaccharides
Rainer Bartels • Helga Bartels • Anna Czabanska • Sylvia Düpow • Dr. Katarzyna Duda • Regina Engel
Kathleen Fischer • Volker Grote • Katharina Jakob • Dr. Uwe Mamat • Olga Neiwert • Gudrun LehwarkYvetot • Kerstin Viertmann • Kathleen Wilke
hygiene hypothesis
Prof. Dr. Otto Holst
MISSION
(AG) were contained in CDE in high concentration. The source
Komaniecka I, Choma A, Lindner B, Holst O. The structure of a
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.
of this AG was fodder, in particular a prevalent grass species
novel neutral lipid A from lipopolysaccharide of Bradyrhizobium
known as Alopecurus pratensis. Treatment of murine dendritic
elkanii containing three mannoses in the backbone. CHEMISTRY
cells (DC) with grass AG resulted in autocrine IL-10 production.
– A EUROPEAN JOURNAL. 2010; 16:2922-2929.
25
most important findings
catalyze the transfer of only a single Kdo residue from CMP-Kdo
Intranasal application of grass AG protected mice from develo-
Peters M, Kauth M, Scherner O, Gelhar K, Steffen I, Werner P,
The chemical structure of the lipid A of the LPS from Bradyrhizo-
to differently modified lipid A acceptors (Fig. 1). The Kdo transfe-
ping atopic sensitisation, allergic airway inflammation and air-
von Mutius E, Holst O, Bufe A. Arabinogalactan isolated from
bium elkanii USDA 76 (slow-growing rhizobia) was established.
rase was capable of utilizing a broad spectrum of acceptor sub-
way hyperreactivity in a mouse model of allergic asthma. This
cowshed dust extract protects mice from allergic airway inflam-
It differed considerably from other lipid A due to the complete
strates, whereas surface plasmon resonance studies indicated a
allergy-protective effect was specific for grass AG since AG from
mation and sensitization, JOURNAL OF ALLERGY AND CLINICAL
lack of negatively charged groups (phosphate or uronic acid re-
high selectivity for the donor substrate. Taken together, the data
gum Arabic and larch did not show allergy-protective properties.
IMMUNOLOGY. 2010; 126:648-656.
sidues), entire replacement of a GlcpN disaccharide backbone
provided evidence that the thermostable WaaA enzyme of A.
by one consisting of 2,3-dideoxy-2,3-diamino-D-glucopyranose
aeolicus is a strictly monofunctional Kdo transferase.
Figure 1:
(GlcpN3N), and by the presence of two long chain fatty acids.
Raoultella terrigena ATCC 33257, recently reclassified from the
Conversion of
Internally, we are collaborating with the Divisions of Immuno-
The GlcpN3N disaccharide was further substituted by three D-
genus Klebsiella, is a drinking water isolate and belongs to a
the tetraacyl-
chemistry (U. Zähringer), Innate Immunity (H. Heine), Cellular
Manp residues, forming together a pentasaccharide. As shown
large group of non-typeable Klebsiella and Raoultella strains.
1,4´-bisphos-
Microbiology (U. Schaible), and Clinical and Experimental Pa-
by 1D and 2D NMR spectroscopy, chemical composition analy-
Using an O-antiserum against a capsule-deficient mutant of this
phate lipid A
thology (E. Vollmer, T. Goldmann).
ses, and high resolution mass spectrometry (ESI FT-ICR MS, MS/
strain, we could show a high prevalence (10.5 %) of the R. terri-
precursor 406
Externally, we are collaborating with the Dipartimento di Chimi-
MS), one D-Manp was linked to C-1 of the reducing GlcpN3N
gena O-serotype among non-typeable, clinical Klebsiella and
(1404.86 u) (A)
ca e Biochimica, Universitá di Napoli Federico II, Naples, Italy
and an α-(1→6)-linked D-Manp disaccharide was located at C-4’
Raoultella isolates. We observed a strong serological cross-reac-
into Kdo-406
(A. Molinaro, C. De Castro), Experimental Pneumology, Ruhr-
of the non-reducing GlcpN3N in α-linkage. Fatty acid analysis
tion with the K. pneumoniae O12 reference strain, indicating that
(1624.92 u) (B)
University Bochum, Germany (A. Bufe), University Children Hos-
internal and external collaborations
identified 12:0(3-OH) and 14:0(3-OH) amide-linked to GlcpN3N.
a large percentage of these non-typeable strains may belong
by the Kdo
pital, LMU, Munich, Germany (E. von Mutius), Chair of Animal
Other constituents were long (ω-1)-hydroxylated fatty acids with
to the O12 serotype, although these are currently not detectable
transferase of
Hygiene, TUM, Munich, Germany, Institute for Laboratory Me-
26 – 33 carbon atoms, as well as their oxo-forms [28:0(27-oxo)
by the K. pneumoniae O12 reference antiserum in use. Therefo-
A. aeolicus.
dicine and Pathobiochemistry, University of Marburg, Germany
and 30:0(29-oxo)]. The 28:0(27-OH) was the most abundant fatty
re, we analyzed the O-polysaccharide (O-PS) structure and ge-
(H. Renz), Haartman Institute, Department of Bacteriology and
acid. As proven by high resolution mass spectrometry techniques
netic organization of the wb gene cluster of R. terrigena ATCC
Immunology, Helsinki, Finland (M. Skurnik), Institute of Microbio-
these long-chain fatty acids created two acyloxyacyl residues
33257, and both confirmed a close relation of R. terrigena and
logy, University of Silesia, Katowice, Poland (J. Radziejewska-Leb-
with the 3-hydroxy fatty acids. Thus, lipid A from B. elkanii com-
K. pneumoniae O12. The two strains possess an identical O-PS,
recht), Department of Genetics and Microbiology, Maria Curie-
prised six acyl residues. One of the acyloxyacyl residues could
lipopolysaccharide core structure, and genetic organization of
Skłodowska University, Lublin, Poland (A. Choma), Institute of
be further acylated at the (ω-1)-hydroxy group by 3-hydroxy bu-
the wb gene cluster. Heterologous expression of the R. terrigena
tyric acid.
wb gene cluster in Escherichia coli K-12 resulted in the WecA-
selected puBlications
The hyperthermophile Aquifex aeolicus belongs to the deepest
dependent synthesis of an O-PS reactive with the K. pneumoniae
Mamat U, Schmidt H, Munoz E, Lindner B, Fukase K, Hanus-
of Bielefeld, Germany (K. Niehaus), Department of Microbiology
branch in the bacterial genealogy. The cells are Gram-negative
O12 antiserum. The serological data presented suggest a higher
zkiewicz A, Wu J, Meredith TC, Woodard RW, Hilgenfeld R,
and Immunology, University of Western Ontario, London Ontario,
with a rather complex cell envelope of a surface protein layer,
prevalence of the O12-serotype among Klebsiella and Raoultella
Mesters JR, Holst O. WaaA of the hyperthermophilic bacte-
Canada (M. A. Valvano), Department of Medicinal Chemistry,
murein, and an outer membrane. Compositional analysis and
isolates than generally assumed.
rium Aquifex aeolicus is a monofunctional 3-deoxy-D-manno-
College of Pharmacy, University of Michigan, Ann Arbor, USA
mass spectrometry of purified A. aeolicus LPS, showing the in-
Extract from cowshed dust (CDE) is a source of immune-modula-
oct-2-ulosonic acid transferase involved in lipopolysaccharide
(R. W. Woodard), Institute of Biochemistry, University of Lübeck,
corporation of a single Kdo residue as an integral component
ting substances which protect from experimental allergic disor-
biosynthesis. JOURNAL OF BIOLOGICAL CHEMISTRY. 2009;
Germany (R. Hilgenfeld).
284:22248-22262.
Nanobiotechnology, University of Natural Resources and Life Sciences, Vienna, Austria (C. Schäffer), Faculty of Biology, University
of the LPS, implicated a monofunctional Kdo transferase in LPS
ders in a mouse model of asthma. In order to identify immune-
biosynthesis of A. aeolicus. Further, heterologous expression of
modulating molecules in CDE from an allergy protective farming
the A. aeolicus waaA gene in a Escherichia coli waaA suppres-
environment, polysaccharides were isolated from CDE and plants
Mertens K, Müller-Loennies S, Stengel P, Podschun R, Han-
Our research has been supported by the Deutsche Forschungs-
sor strain resulted in synthesis of lipid IVA precursors substituted
by chromatography and precipitation with specific reagents, and
sen DS, Mamat U. Antiserum against Raoultella terrigena
gemeinschaft (SFB-TR 22, project A2, FOR 585; MA 1408/2-1
with one Kdo sugar. When highly purified WaaA of A. aeolicus
then characterized by NMR-spectroscopy. The allergy-protective
ATCC 33257 identifies a large number of Raoultella and Kleb-
and MA 1408/2-2), the Europenian Union (GALTRAIN, GABRIEL)
was subjected to in vitro assays using mass spectrometry for
potential of the isolated polysaccharides was tested in a mouse
siella clinical isolates as serotype O12. INNATE IMMUNITY.
and industry.
detection of the reaction products, the enzyme was found to
model of asthma. We demonstrated that plant arabinogalactans
2010; 16:366-380.
Grant support
Dept. Molecular Infection biology • Structural Biochemistry
Such DC were not able to induce an allergic immune response.
Immunochemistry
Structural analysis of proinflammatory bacterial compounds
Dr. Christian Alexander • Birte Buske • Dr. Nicolas Gisch • Heiko KäSSner • Brigitte Kunz • PD Dr. Buko
Lindner • Helga Lüthje • Hermann Moll • Wiebke Schnoor • Ursula Schombel • Simone Thomsen • Nicole
Zehethofer
PAMPs
Lipidomes
MISSION
methods that allow detailed characterization of the complex
National
The Division of Immunochemistry aims to identify the structure of various BACTERIAL compounds known as pathogen-associated molecular patterns, (PAMPs). Based on a profound structure-function relationship, we aim to contribute to the establishment of improved new treatments, which are necessary to prevent and control pro-inflammatory and harmful reactions in the host.
lipid composition obtained under different experimental
Schmidt (Konstanz), Hammerschmidt (Greifswald), Bekeredjian-
conditions from tissue (lung epithel, intestinal endothel),
Ding, (Heidelberg), Meyer (MPI Berlin), Bufe (Bochum), Scheel
different cell types, subcellular compartments, membranes and
(Halle), Grötzinger, Rosenstiel, Saftig, Schröder, Röder (Kiel),
membrane microdomains (signaling platforms, rafts).
König, Hübner (Lübeck).
International
most important findings
(HIC) and analyzed by FT ICR-MS and NMR, and biologically
selected puBlications
Grzesiek, Cornelis (Basel), Reichhart (Strasbourg), Beutler, (La
Analysis of LPS and lipid A by NMR and MS
evaluated (collaboration with Prof. A.J. Ulmer, Division of Innate
1. Shin H, Mally M, Meyer S, Fiechte C, Paroz C, Zähringer
Jolla), Philpott (Toronto), Puzo, Gacias-Alles (Toulouse) Knirel
C,15N-labelled LPS by
Immunity). We found that not TLR2 but another receptor of the
U, Cornelis GR. Resistance of Capnocytophaga canimorsus
nuclear magnetic resonance (NMR) in a water mimetic [annual
innate immune system, the lectin pathway of the complement,
to killing by human complement and polymorphonuclear
leukocytes. Infect Immun 2009; 77: 2262-2271.
After we could successfully analyze
13
report 2008] we succeeded in the establishment of an analytical
has gained increasing attractivity. With the help of synthetic
and preparative RP-HPLC-technique, which enabled us for the
LTA we obtained first evidences that this receptor is indeed the
first time to prepare highly purified LPS and lipid A, which were
pathogen recognition receptor (PRR) for LTA.
our new 700 MHz NMR ultrashielded magnet in December 2009,
Synthesis of biotinylated muramyl tripeptides with NOD2-
equipped with cryo probe, we are now able to run NMR spectra
stimulating activity Muramyl di- and tripeptides are
antibody S60-4-14 reveals diagnostic potential in the
with 6 fold increase higher sensitivity, thus being able to analyze
supposed to be activators of the innate immune system by
identification of Pseudomonas aeruginosa in lung tissues
natural and purified LPS, glycolipids and lipid A substructures
stimulating the NOD2 receptor.
of cystic fibrosis patients. Eur J Cell Biol. 2010; 89 :25-33.
without expensive C, N labelling.
To provide tools for the clarification of the molecular mechanism
13
15
of this activation we isolated different UDP-muramyl tripeptides
Länger F, Meyer R, Welte T, Zähringer U. Monoclonal
3. Pedersen CM, Figueroa-Perez I, Lindner B, Ulmer AJ,
(Lys- and DAP-type) from bacteria and modified them chemically
Zähringer U, Schmidt RR. Total synthesis of lipoteichoic
to obtain biotinylated derivatives of natural occurring muramyl
acid of Streptococcus pneumonieae. Angew Chem Int Ed
tripeptides. All modified compounds retained their ability to activate
Engl. 2010; 49: 2585-2590.
NOD2 in a cell-based test system and will therefore be suitable
tools to identify the appropriate pattern recognition receptor(s).
4. Pedersen CM, Figueroa-Perez I, Boruwa J, Lindner B,
Ulmer AJ, Zähringer U, Schmidt RR. Synthesis of the
Detailed lipid analysis of complex biological samples
core structure of the lipoteichoic acid of Streptococcus
Lipids influence biological processes such as signal transduction.
pneumoniae. Chemistry. 2010; 16: 12627-12641.
Therefore, we established sensitive LC- FT ICR-MS, MS/MS
5. Schmidt RR, Pedersen CM, Qiao Y, Zähringer U. Chemical
synthesis of bacterial lipoteichoic acids: an insight on its
biological significance. Org Biomol Chem. in press.
6. Zehethofer N, Scior T, Lindner B. Elucidation of the
Figure 1: First 1H,13C HSQC NMR spectrum E. coli-type lipid A (LA) in
fragmentation pathways of different phosphatidylinositol
water using small DHPC-d40. black: 13C,15N-labelled LA, red: hexa-acyl
phosphate species (PIPx) using IRMPD implemented on a
LA after HPLC purification, orange: DHPC-d40 signals.
FT-ICR MS. Anal Bioanal Chem. 2010; 398: 2843-2851.
Synthesis, purification, structural and biological analysis
of natural and synthetic Lipoteichoic acid (LTA) from
internal and external collaborations
Staphylococcus aureus and Streptococcus pneumoniae.
Inhouse
Various LTAs of S. aureus and S. pneumococcus-type have been
Grant support
DFG (Za-149/6-1), CLUSTER OF EXCELLENCE (IRN G TP2), SFB
2. Schmengler K, Goldmann T, Brade L, Sánchez Carballo
PM, Albrecht S, Brade H, Kosma P, Sahly H, Hauber HP,
analyzed by high-resolution FT ICR-MS. After the installation of
(Moscow), Raina, Klein (Gdansk).
Heine, Ulmer, Goldmann, Holst, Mamat, Reiling, Hauber,
synthesized in the group of Prof. R.R. Schmidt (University of
Figure 2: LPS induced changes of ceramides in cellular signaling
Gutsmann,
Schromm,
Grassl,
Schaible,
Konstanz), purified by hydrophobic interaction chromatography
platforms (DRMs).
F.Petersen, A.Petersen, Fehrenbach.
Müller-Loennies,
TR 22 (Z01).
27
Dept. Molecular Infection biology • Immunochemistry
Prof. Dr. Ulrich Zähringer
Biophysics
Antimicrobial peptides
Lipid membrane
Glycolipid
PD Dr. Jörg Andrä • Prof. Dr. Klaus Brandenburg • Sabine Dabelstein • Sabrina Groth • Nina Halbrock
Christine Hamann • Yani Kaconis • Max Koistinen • Annika Kopp • Ina Kowalski • Kerstin Stephan
Pathogenicity factor
Signal transduction
MISSION
presence and/or shape of the cellular surface (glyco)-structures
G. Martinez de Tejada, Universidad de Navarra, Pamplona
The function of lipids and lipid membranes and their interaction with peptides and proteins is very important in the context of infections. We use a number of biological and biophysical techniques to characterize bacterial and human immune cell membranes and in particular reconstituted membranes mimicking their natural examples. The goal is to establish and to use model membranes which are simple enough to allow a molecular analysis of the underlying mechanisms and to be as close to the natural system to allow a prediction of their biological behavior. The biomedical focus is directed towards (a) the organization and properties of membranes composed of phospho- and glycolipids, (b)
the function of antimicrobial peptides (AMPs) of the innate immune system, and (c) the elucidation of membrane-associated signal transduction pathways.
have a significant impact on peptide-cell interaction. We compre-
N. Tanic and J. Bankovic, Institute for Biological Research Bel-
hensively analyse peptide interactions with human cancer cell
grade, Serbia
lines, which naturally or artificially differ in the surface exposition
of negatively charged glycans or in the thickness of the glycocalyx. The core instrumentation of this application is a metabolic
chip biosensor, which enables the assessment of the dynamics of important cell physiological parameters. To correlate the
biological activities of peptides with the presence and chemical
structure of cellular surface molecules, the biological data are
complemented by peptide-membrane interaction studies.
most important findings
AMPs investigated and their interaction with pure lipid matrices
Bacterial infections are still one of the major threats to human
has been found. Thus, as a first step, the permeabilization of the
health worldwide. Bacterial pathogens frequently cause severe
lipid membrane is an essential prerequisite for bacterial killing.
Andrä J, Hammer M, Grötzinger J, Jakovkin I, Lindner B,
diseases not only as primary agents, but also subsequent to
Based on our findings we were able to synthesize first genera-
Vollmer E, Fedders H, Leippe M, Gutsmann T. Significance of
pathologies caused by other agents. This fact is due, at least in
tion peptides with improved activities. In addition to the pore for-
the cyclic structure and of arginine residues for the antibacterial
part, to the increasing occurrence of bacterial resistance to anti-
mation we investigated further mechanisms including membrane
activity of arenicin-1 and its interaction with phospholipid and li-
biotics. In turn, the spread of antibiotic-resistant clones is greatly
fusion or aggregation induced by hydramacin and membrane
popolysaccharide model membranes. BIOLOGICAL CHEMISTRY
enhanced by the improper use of antibiotics and by the massive
encapsulation induced by the C-reactive protein from Limulus.
2009 Apr;390(4):337-349.
Development of new polypeptides as general microbiocides
Hammer MU, Brauser A, Olak C, Brezesinski G, Goldmann
Reconstitution Systems
We have developed new peptides, which were directed to com-
T, Gutsmann T, Andrä J. Lipopolysaccharide interaction is de-
To mimic bacterial and immune cell membranes we established
bat against the Gram-negative sepsis in that the main pathoge-
cisive for the activity of the antimicrobial peptide NK-2 against
new reconstitution systems. The bilayers were composed of phos-
nicity factor LPS is neutralized by the peptides. Thus, these were
Escherichia coli and Proteus mirabilis. BIOCHEMICAL JOURNAL
pholipids, lipid II, and lysyl-PG to mimic the membrane of Gram-
called SALP (synthetic anti-LPS peptides). They were shown to
2010 May;427(3):477-488.
positive bacteria, and phospholipids and cholesterol to mimic
effectively block the inflammation reaction in vitro as well as the
Figure 1:
In vivo mouse
model of LPS
treated mice
protected by
the AMP LPep
19-2.
selected puBlications
application of antibiotics in animal husbandry.
Figure 2:
Mycobacterial
trehalose-dimycolate domains
visualised in
giant unilamellar vesicles
(top) and in
solid supported
bilayers by AFM
(bottom).
the cytoplasmic membrane of eukaryotic cells. Furthermore, we
systemic inflammation in vivo (mouse model of endotoxemia and
Gutsmann T, Razquin-Olazarán I, Kowalski I, Kaconis Y,
reconstituted the outer membrane of Gram-negative bacteria as
of infection by bacteria) as well as to inhibit Gram-positive pneu-
Howe J, Bartels R, Hornef M, Schürholz T, Rössle M, Sanchez-
Grant support
asymmetric planar lipid bilayer and also as asymmetric lipo-
monia in combination with an antibiotic. Additionally, some of
Gómez S, Moriyon I, Martinez de Tejada G, Brandenburg
German ministry BMBF (grant 01GU0824; ‘Therapy of infectious
somes with one side composed of lipopolysaccharides (LPS), the
the peptides were shown to considerably inhibit virus replication
K. New antiseptic peptides to protect from endotoxin-mediated
diseases with special regard to bacterial sepsis’)
other of phospholipids. In first attempts we have been successful
in human cells, such as those of the human immunodeficiency vi-
shock. ANTIMICROBIAL AGENTS AND CHEMOTHERAPY 2010
Deutsche Forschungsgemeinschaft (GU 568/4-1, ‚Biophysical
to mimic the mycobacterial wax layer by using lipid matrices
rus (HIV), Herpes simplex viruses I and II, hepatitis B, seasonal in-
Sep;54(9):3817-24.
investigations into the interactions between antimicrobial pep-
containing the glycolipid trehalose dimycolate (TDM).
fluenza H3N2, Dengue fever virus, and classical swine fever virus.
The mechanisms of the anti-septic action and of virus replication
internal and external collaborations
AN 301/5-1 ‚Significance of cellular surface factors for the selec-
Function of Antimicrobial Peptides (AMPs)
differ: For the anti-septic activity, a direct binding to the bacterial
Internal cooperation:
tive cytotoxicity of antimicrobial peptides (AMPs) against cancer
The AMPs we have investigated within the last years differ in
pathogens take place, but also a binding to cell surface recep-
Norbert Reiling, Ulrich Schaible, Andreas Frey, Otto Holst, Tor-
cells‘)
their structure and activity. Starting with our hypothesis that lipid-
tors such as CD14 and TLR4.
sten Goldmann and Ekkehard Vollmer, Andra Schromm.
Leibniz-Graduiertenschule ‚Modellsysteme für Infektionskrank-
External cooperation:
heiten‘
Clinique La Prairie, Clarens, Switzerland (‚Biophysical investiga-
and peptide-specific properties are responsible for the sensitivity
29
tides of the epithelial defense and microbial cell envelopes’;
or resistance of certain bacterial strains, we have characterized
Anti-cancer activity of AMPs
R. Schmitz-Streit, University of Kiel
the interactions between the AMPs and reconstituted lipid mem-
The objectives of another project is to gain an understanding of
T. Lindhorst, University of Kiel
tions into the interaction of endotoxin with hemoglobin and part
branes. For our investigations we have used various AMPs in
the dynamics of the interactions of antimicrobial peptides with
M. Hornef, Medical School Hannover
structures‘).
their natural form as well as synthetic derivatives thereof. An
human cancer cells and the molecular basis for differential cy-
T. Schürholz, Medical School Aachen
almost perfect correlation between the biological activity of the
totoxicity and mode of action based on the hypothesis that the
U. Maus, Medical School Hannover
Dept. Molecular Infection biology • Biophysics
Prof. Dr. Thomas Gutsmann
Immunobiophysics
Macrophage activation and regulation
Sabrina Groth • Susanne Keese • Franziska Kopp • Isabelle Mehrez-Koch • Irina von Cube • Sonja Winkler
Immune modulators
Biophysical techniques
Reconstituted model membranes
PD Dr. Andra Schromm
MISSION
reconstitution models, to elucidate the underlying mechanisms.
lipoprotein from Mycobacterium tuberculosis. INNATE IMMUNITY
Our team investigates 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 matrices, confocal microscopy, high sensitivity spectroscopy, and modern immunological and cell biological techniques.
In this interdisciplinary project, we also acquire and employ new
2010, 16(4):213-25.
techniques to tackle structural questions in biomedical research.
between Lipid A and Serum Proteins. ADVANCES IN EXPERI-
Microbial recognition by lectins represents a major part of the
MENTAL MEDICINE 2010 ; 667:39-51.
humoral innate immune defense against bacterial infections.
the molecular mechanisms of immune recognition of the Mtb-LP
The soluble surfactant proteins A (SP-A) and D, secreted by
Brandenburg K, Schromm AB, Gutsmann T. Endotoxins: rela-
Lipid mediated signaling
are tailored to the ambient conditions of the lung.
type-II pneumocytes, are the central components of lung innate
tionship between structure, function, and activity. SUBCELLULAR
immunity. SP-A and SP-D elicit immune-modulatory functions on
BIOCHEMISTRY 2010 ; 53:53-67.
interdisciplinary characterization of the biophysical and
Investigations into the cellular function of the acute-phase
alveolar macrophages, the primary resident innate immune
molecular basis of the initiation and regulation of the
protein LBP in antimicrobial immunity
cell in the lung. Pulmonary diseases caused by Gram-negative
internal and external collaborations
inflammatory immune response to bacterial membrane
After the decipherment of the genetic code, the major
bacteria are the leading cause of mortality from infectious
Internal collaborations:
components. Endotoxin (lipopolysaccharide, LPS), the main
challenge of biomedical research is the elucidation of the
diseases. Effective treatment of these diseases is a major
Helmut Brade, Division of Medical and Biochemical Microbiology
amphiphilic component of the outer layer of the outer membrane
structure, function and dynamics of the critical players.
task to improve medical care for the rising number of elderly
Thomas Gutsmann, Division of Biophysics
of Gram-negative bacteria, is among the most potent stimuli
Acute-phase proteins are a central part of the innate immune
individuals und immunocompromised patients. Modulation of
Norbert Reiling, Division of Microbial Interface Biology
of the innate immune system. In patients with Gram-negative
defense against infections. They recognize microbial associated
the host’s innate immune response is an attractive target to
Cordula Stamme, Division of Cellular Pneumology
infections it is a central mediator in the development of sepsis
molecules and aid in initiating the humoral and cellular immune
improve clinical outcome, especially considering the increasing
Ulrich Zähringer, Division of Immunochemistry
and septic shock, pathophysiological conditions with a lethality
response. Lipopolysaccharide-binding-protein (LBP) specifically
numbers of antibiotics-resistant pathogens. Aiming to elucidate
External collaborations:
of about 50 %. Currently, there is no therapy available to
binds structures of Gram-positive and Gram-negative bacteria,
the basis of immune-modulation conferred by SP-A, we
Manfred Roessle, HASYLAB c/o DESY, Hamburg
improve the outcome of sepsis and septic shock. We investigate
mediates the transport of these molecules to cellular receptors
investigate the physico-chemical consequences of SP-A – LPS
Michael Steinert, Technical University, Braunschweig
lipopolysaccharides and structural mimetics to define the
thereby initiating transmembrane signaling. We have recently
interaction on the molecular level. We were able to observe
Andreas Tholey, Proteomics Unit, Christian-Albrechts-University
molecular connection to agonistic and antagonistic activity in
described, that besides the soluble protein LBP also a cell-
Figure 2:
Kiel
the human immune system.
associated membrane form of LBP is important for the
SP-A induces
Karl-Heinz Wiesmüller, University Tübingen, EMC microcollections
Besides LPS, lipoproteins and lipopeptides represent a major
activation of the immune response. Our current work on LBP
physico-chemi-
EISAI Research Institute, Andover, USA
group of bacterial virulence factors with strong immune-
is aimed at elucidating the structure, function and dynamics of
cal changes
modulatory capacity, including beneficial effects in infectious
this membrane-associated LBP. We are approaching this task
in bacterial
Grant support
diseases by stimulation of the innate and adaptive immune
by obtaining data on the one hand from experiments using
lipopolysaccha-
DFG SCHR 621/2-3 (A.Schromm): „Immunerkennung Gram-
response. To learn more about the mechanisms of lipoprotein
primary immune cells as well as genetically modified cell lines
ride.
negativer Erreger: Untersuchungen zur molekularen Struktur
recognition in infectious diseases, we investigated a synthetic
Figure 1:
und Funktion des LPS-Rezeptor-Komplexes“ Emmy-Noether
lipopeptide
mimetic
of
the
19-kD
lipoprotein
from
Localization
Programms der DFG (until 10/2009)
Mycobacterium tuberculosis, an intracellular pathogen that
of membrane
DFG SCHR 621/3-1 (A. Schromm): „Immunregulation in der
is a major health threat worldwide. The 19-kDa lipoprotein
LBP on human
Lunge durch pulmonale Collectine”
from M. tuberculosis has been assigned an important role
macrophages.
Leibniz Graduate School „Modell systems for infectious
diseases“: “Investigations into the cellular function of the acute-
in the induction of an antibacterial immune response in host
macrophages. Investigation of the three-dimensional geometry
that distinct functional groups of the LPS molecule are affected
phase protein LBP in antimicrobial immunity” (A.B. Schromm und
of the synthetic 19-kDa M. tuberculosis-derived lipopeptide (Mtb-
by SP-A binding, leading to specific conformational changes
T. Gutsmann)
LP) and the preference for membrane intercalation could provide
that can explain the reduced bioactivity of LPS bound to SP-A.
an explanation for the biological activities of the mycobacterial
selected puBlications
LP. Surprisingly, we found that – in contrast to what we observe
for other lipopeptides – activation of macrophages to induce
and on the other hand from well defined reconstituted model
Schromm AB, Reiling N, Howe J, Wiesmüller KH, Roessle
pro-inflammatory mediators in response to the Mtb-LP was
membranes. The basic concept is to correlate the findings from
M, and K Brandenburg. Influence of serum on the immune
strongly enhanced under serum-free conditions, suggesting that
the complex biological system with those from well defined
recognition of a synthetic lipopeptide mimetic of the 19-kD
Dept. Molecular Infection biology • Immunobiophysics
Andrä J, Gutsmann T, Müller M, Schromm AB. Interactions
Immune regulation in the lung by pulmonary collectins
most important findings
The Division of Immunobiophysics is focused on the
31
Molecular Inflammation Medicine
anti-infective therapies
Tuberculosis
TBornotTB
Martina Ackermann • Dr. Sahar Aly • Dr. Evelin Grage-Griebenow • Silvia Maass • Dr. Judith Petersen
Christina Trabandt • Dr. Kerstin Walter
in vivo test station
cluster of excellence
MISSION
a weapon against mycobacterial replication. The purpose of
Franz-Christoph Bange, Medical Microbiology, Medizinische
The division investigates the pathogenesis of tuberculosis in mouse models mimicking human immunopathology. These models serve as a preclinical test station to (i) explore mechanisms of mycobacterial virulence and persistence, and (ii) validate discoveries in genetic susceptibility and novel anti-infective therapies. The division is part of the translational TB Center Borstel.
the adaptive immune response is therefore not necessarily to
Hochschule Hannover
make a granuloma, but to activate cells within a granuloma for
Klaus Schughardt, Helmholtz Center for Infection Research,
mycobacteriocidal or -static function.
Braunschweig
An evolutionary perspective takes into account the mutual
Michael Niederweis, Microbiology, University of Birmingham/
shaping of the tissue microenvironment, which concurrently
AL (USA)
Peter Sander, Medical Microbiology, University of Zürich (CH)
most important findings
MBL1/2 differentially affected the outcome of aerosol infections
allows propagation and transmission of Mtb, yet restricts tissue
Our laboratory has long been skeptical about the relevance
with M. tuberculosis and M. africanum in mice. We hypothesize
damage to safeguard survival of the host. Mtb needs an exit
of innate immunity in the response to M. tuberculosis (Mtb)
that MBL binding may facilitate the uptake of M. africanum by
route from the granuloma if it is to infect and multiply in other
infection in vivo. In vitro, Toll-like receptors (TLR) 2, 4 and 9 as
macrophages, thereby promoting infection and that selection by
human beings (the latter being the only relevant host). Foamy
well as NOD-like receptor 2 critically determine macrophage
TB may have favored the spread of functional MBL deficiencies
macrophages have recently attracted attention as potentially
responses to Mtb infection. However, in low-dose experimental
in regions endemic for M. africanum.
significant players in this scenario. It has long been clear that
murine tuberculosis, single or multiple deficiencies in TLRs 2,
Mechanisms by which Mtb adapts to acidic environments are
bacterial lipids have a significant role in shaping the tissue
4, 9 or NOD2 have little, if any, impact on early mycobacterial
poorly understood. OmpATb, the only known outer membrane
response at the infection site. More specifically, trehalose-
growth containment, granuloma formation and survival. We
protein required for Mtb virulence, is not a general porin (coop.
dimycolate, which accumulates in necrotic regions of mouse
analyzed the relevance of NALP3, one component of the
M. Niederweis, Birmingham/AL). However, chemical analysis of
granulomas, is specifically recognized by macrophages via the
danger-signaling inflammasome, for (i) Mtb-induced cytokine
culture filtrates showed that proteins encoded by the ompATb
monocyte-inducible C-type lectin (Mincle). Necrosis as such may
secretion in vitro and in vivo, (ii) restriction of Mtb replication in
operon, Rv0899, Rv0900 and Rv0901, are involved in generating
therefore also occur as a consequence of Mtb-derived factors
infected organs and (iii) granuloma formation. In the absence of
a rapid ammonia burst. Addition of exogenous ammonia and
interacting with innate immune cells. When viewed in this
functional NALP3, there was no IL-1beta and IL-18 production in
expression of the ompATb operon restored rapid ammonia
context of the life cycle of Mtb, granulomas make better sense.
Figure 1:
Kinetic in vivo
imaging system
for Mtb-infected
mice under
BSL3 conditions
Mtb-infected dendritic cells and macrophages in vitro, whereas
secretion, medium pH neutralization and concomitant growth
secretion of IL-1alpha, IL-12p40 and TNF remained unaffected.
of the DompATb mutant demonstrating that ammonia secretion
selected puBlications
Figure 2:
After three weeks of infection, NALP3-deficient as well as IL-
mediated by Rv0899, Rv0900 and Rv0901 is the main mechanism
Ehlers S, Kaufmann SH. Infection, inflammation, and chronic
Foamy macro-
18-deficient mice were as capable as wildtype mice of
by which Mtb neutralizes acidic environments in vitro. Infection
diseases: consequences of a modern lifestyle. Trends Immunol
phages (oil red
restricting Mtb loads at a plateau level within well-differentiated
experiments with two DompATb Mtb mutants revealed that the
2010; 31(5):184-90 (Review)
stain) surroun-
granulomas. In conclusion, despite its involvement in cytokine
ompATb operon was not required for full virulence in mice.
Ehlers S. TB or not TB? Fishing for Molecules Making Permissive
ding caseating
processing, NALP3 is not essential for induction of protective
These results show that genes within the ompATb operon are
granulomas. Cell Host Microbe 2010;7(1):6-8 (Review)
necrosis in
immunity to Mtb.
necessary for rapid ammonia secretion and adaptation of Mtb
Walter K, Hölscher C, Tschopp J, Ehlers S. NALP3 is not
experimental TB
Structural variants of the Mannose Binding Lectin (MBL) cause
to acidic environments, but that their deletion is not sufficient for
necessary for early protection against experimental tuberculosis.
granuloma
quantitative and qualitative functional deficiencies, which are
growth attenuation in vivo.
Immunobiology 2010; 215(9-10):804-11.
associated with various patterns of susceptibility to infectious
The group actively contributes to redefining the role of
Ehlers S. DC-SIGN and mannosylated surface structures of
diseases. Genetic MBL variants in 2010 Ghanaian patients with
the granuloma in TB. Mycobacteria are phagocytosed by
Mycobacterium tuberculosis: a deceptive liaison. Eur J Cell Biol
pulmonary tuberculosis (TB) and 2346 controls were determined
macrophages which support their intracellular replication. It is
2010;89(1):95-101 (Review)
(coop. C.G. Meyer, Bernhard-Nocht-Institut, Hamburg), and the
therefore in the best interest of the mycobacterium to stimulate
Ehlers S. Lazy, dynamic or minimally recrudescent? On the
mycobacterial isolates of the patients were characterized at the
innate inflammatory responses to induce the accumulation of
elusive nature and location of the mycobacterium responsible
Grant support
same time (coop. S. Niemann, FZB). Assuming a recessive mode
unactivated macrophages in order to further its own replication.
for latent tuberculosis. J Infection 2009; 37(2):87-95 (Review)
DFG/BMBF: Cluster of Excellence Inflammation at Interfaces,
of inheritance, a negative association between TB and the MBL2
Granulomas form even in the absence of specific immunity
G57E variant (odds ratio 0.60) and the corresponding LYQC
(for example in SCID mice). It is a common misconception that
internal and external collaborations
BMBF: Network “Pulmonary Tuberculosis – Host and Pathogen
EXC 306 (2007-2012)
haplotype (Pcorrected 0.007) was identified. This applied, however,
granulomas, by their very existence, “wall off” mycobacteria;
TB Center Borstel
Determinants
only to TB caused by M. africanum but not to TB caused by M.
rather, they provide the fertile soil on which mycobacteria thrive.
Christian G. Meyer, Bernhard-Nocht-Institut for Tropical Medicine,
(TBornotTB)”, (2007-2014)
of
Resistance
tuberculosis. In vitro, M. africanum isolates bound recombinant
Only the T cell mediated activation of cells assembled in the
Hamburg
BMBF:
human MBL (and recombinant mouse MBL1 and MBL2) more
granuloma, be it via interferon-gamma, tumor necrosis factor
Ulrich Maus, Experimental Pneumology, Medizinische Hoch
tuberculosis” (2007-2011)
efficiently than did isolates of M. tuberculosis. A deficiency in
or granulysin, is actually capable of turning the granuloma into
schule Hannover
Network
“Short
and
course
Disease
Progression
chemotherapy
against
33
Dept. Molecular Infection biology • Molecular Inflammation Medicine
Prof. Dr. Stefan Ehlers
Infection Immunology
Alternatively activated
macrophages
Tuberculosis
Chagas
Dr. Jochen Behrends • Julia Böhme • Dr. Hanna Erdmann • Lisa Heitmann • Alexandra Hölscher • Kristian Holz • Gabriele Röver • Erik Schmok • Jan Christian Sodenkamp • Tanja Sonntag • Kerstin Traxel •
Johanna Volz
TH17
MISSION
LysMcreSOCS3loxP/loxP mice. Our data indicate a new pivotal
kemper C, Krell H, Libert C, Lund L, Frey O, Holscher C, Iwakura Y,
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 and Chagas disease.
role of SOCS3 for macrophage activation, enabling efficient
Ghilardi N, Ouyang W, Kamradt T, Sabat R, Liesenfeld O.
killing of intracellular mycobacteria. Together, alternative
Interleukin (IL)-23 mediates Toxoplasma gondii-induced
macrophage activation strongly contributes to susceptibility
immunopathology in the gut via matrixmetalloproteinase-2
and pathology in Mtb infection. Further studies will unravel
and IL-22 but independent of IL-17. JOURNAL OF EXPERI
Hence, suppression of classical activation in alternatively
the underlying mechanisms by which Mtb subverts efficient
MENTAL MEDICINE 2009; 206: 3047-3059.
Tuberculosis (TB) infection starts with the inhalation of infectious
activated macrophages contributes to susceptibility and
intracellular killing in these macrophages.
Mycobacterium tuberculosis (Mtb) into the lung and the uptake
pathology during Mtb infection. With this respect, suppressor
of bacteria by resident alveolar macrophages. This host-cell/
of cytokine signalling 3 (SOCS3) is a feedback inhibitor
Though IL-17A-producing TH17 cells are potent inducers of tissue
P, Lang R, Hölscher C. Autocrine IL-10 induces hallmarks
bacilli interaction leads to the induction and secretion of a
of IL-6 signalling pathways in macrophages. To assess
inflammation and have been associated with several chronic
of alternative activation in macrophages and suppresses
most important findings
Schreiber T, Ehlers S, Heitmann L, Rausch A, Mages J, Murray
variety of cytokines and chemokines. Via this process, other
the putative inhibitory capacity of SOCS3 on macrophage
and autoimmune inflammatory diseases, the main function of
antituberculosis effector mechanisms without compromising T
immune cells are recruited to the site of infection leading to
activation during infection with Mtb, macrophage-specific
TH17 cells appears to be the clearance of pathogens, which
cell immunity. JOURNAL OF IMMUNOLOGY 2009; 183: 1301-1312.
the formation of granulomas in order to contain mycobacterial
SOCS3-deficient
utilized.
are not appropriate affected by TH1 or TH2 cells. To evaluate
growth. However, in some patients the infection reactivates
Aerosol infection of LysMcreSOCS3loxP/loxP mice resulted in an
the role of IL-17A and its protective role during infectious
internal and external collaborations
and these granulomas may necrotize. Mechanisms leading to
utterly
remarkably
diseases, we analyzed IL-17A -/- mice after infection with Mtb or
Ehlers S, Division of Molecular Inflammation Medicine, Research
granuloma necrosis, the hallmark of human TB pathology, are
higher bacterial loads, enhanced pulmonary pathology and
the protozoan parasite Trypanosoma cruzi, the causative agent
Center Borstel; Schaible U, Division of Cellular Microbiology,
not understood. In a human study group, a structural variant of
of human Chagas’ disease. Interestingly, infected IL-17A -/- mice
Research Center Borstel; Thye T, Meyer CG, and Horstmann
the IL-4 receptor-alpha (IL-4Ra) was found to be significantly
efficiently generated interferon-gamma (IFN-g)-producing TH1
R, Department of Molecular Medicine, Bernhard-Nocht-
associated with the cavity size of human granulomas. However,
cells and IFN-g-dependent effector responses but were not
Institute for Tropical Medicine, Hamburg, Germany; Jacobs T,
in Mtb-infected mice IL-4 and IL-13 are only moderately induced
able to control infection. Hence, IL-17A may directly mediate
Bernhard-Nocht-Institute for Tropical Medicine, Department
and necrotizing granulomas are not apparent. To address
macrophage effector mechanisms against infection with
of Immunology, Hamburg, Germany; Lang R, Molecular
the question how these T helper (TH) 2 cytokines may alter
intracellular pathogens. Further in vitro analysis revealed IL-
Medicine, University Hospital Erlangen, Germany; Rose-John
mice
(LysMcreSOCS3loxP/loxP)
immunocompromised
phenotype
were
with
the course of disease, IL-13-overexpressing mice (IL-13 ) were
17A to be able to directly stimulate anti-microbial activities
S, Department of Biochemistry, Christian-Albrechts-University
infected with Mtb. The most important finding was that these
in macrophages leading to the eradication of internalized
Kiel, Germany; Müller U, University of Leipzig, Germany;
tg
mice strongly resembled human TB pathology developing
pathogens. Taken together, our results suggest that IL-17A is
Brombacher F, International Centre for Genetic Engineering and
necrotizing granulomas, which were surrounded by a fibrous
necessary for host protection by stimulating anti-microbial
Biotechnology, University of Cape Town, South Africa.
rim and a layer of arginase-1-expressing alternatively activated
mechanisms in macrophages.
Grant support
macrophages.
Figure 2: Arginase-1 is expressed by macrophages in lungs from
Mtb-infected IL-13tg mice. Immunohistochemical staining of arginase-1
(green staining) and CD63 (red staining) was performed in kryo-sections
of lungs from day 98 infected mice. Double-positive cells were predominantly found below the fibrous rim of necrotizing granulomas.
premature death compared to SOCS3
loxP/loxP
In summary, our studies so far revealed that TH2 and TH17
DFG HO 2145/4-3: “Regulation of inflammatory and protective
immune
macrophage immune responses through Stat3-dependent
responses
regulate
or
stimulate
anti-microbial
macrophage effector responses after infection. Currently,
signalling events”.
we are trying to disclose macrophage-specific downstream
SFB 415, project C10: “Cell type-specific mechanisms mediated
mechanisms during infectious diseases in vivo by the generation
by IL-27/WSX-1 during inflammatory immune responses”.
and analysis of conditional gene-deficient mice.
BMBF: Nation-wide Collaborative Grant: “Pulmonary Tuberculosis
– host and pathogen determinants of resistance and disease
mice. Detailed
analysis revealed that the increased susceptibility to Mtb
selected puBlications
infection was accompanied by an enhanced development
Sodenkamp J, Behrends J, Förster I, Müller W, Ehlers S,
Cluster of Excellence “Inflammation at Interfaces”: Integrative
of alternatively activated macrophages. This alternative
Hölscher C. gp130 on macrophages/granulocytes modulates
Research Network F “Cytokine signalling via gp 130”, project 4
activation of macrophages led to an increased expression
inflammation during experimental tuberculosis. EUROPEAN
“Infection driven inflammation models”.
of arginase-1, thereby providing a niche for Mtb to grow.
JOURNAL OF CELL BIOLOGY 2010 Dec (in press).
Figure 1: Enhanced pulmonary pathology in Mtb-infected LysMcreSOCS-
Treatment of mice with anti-IL-6 antibody during infection
3loxP/loxP mice. Representative photography of lungs isolated 28 days
largely restored anti-mycobacterial effector mechanisms.
Munoz M, Heimesaat M, Danker K, Struck D, Lohmann U,
after infection with 100 CFU Mtb.
IL-6 is therefore involved in promoting susceptibility in
Plickert R, Bereswill S, Fischer A, Dunay I, Wolk K, Lodden
progression.” Workpackage E (Animal models).
35
Dept. Molecular Infection biology • Infection Immunology
Dr. Christoph Hölscher
Microbial Interface Biology
Mycobacterium tuberculosis
pathogen variability
Julius Brandenburg • Lisa Dost • Svenja Kröger • Jan Neumann • Kolja Schaale • Jeanette Schwarz
Katrin Seeger • Christine Steinhäuser
in vitro drug testing
phagosome isolation
macrophage Signalling
MISSION
Blot analysis identified known and novel phagosomal proteins.
Gerdes, Ulrich Schaible, Ulrich Zähringer), Niels Röckendorf,
The main focus of the division, which was established in late 2008, is the detailed molecular characterization of the interaction between pathogenic mycobacteria and their target cells, the macrophages.
This includes (i) creating a molecular map of the M. tuberculosis phagosome, (ii) deciphering of M.
tuberculosis-induced signaling pathways, (iii) associating the genetic diversity of the M. tuberculosis complex strains with distinct macrophage response profiles and (iv) testing of novel anti-mycobacterial agents in primary macrophages.
The newly developed method represents a powerful tool for the
Andreas Frey, Mucosa Immunology;
detailed molecular analysis of the intracellular host-pathogen
Stefan Schütze, Andreas Tholey, University of Kiel; Jan Rupp, Univer-
interface, facilitates comparative studies across the kingdom of
sity of Lübeck; Karl Heinz Wiesmüller, EMC microcollections, Tübin-
intracellular pathogens, and is useful for identifying microbial
gen; Eberhard Krause, FMP, Berlin; Ger van Zandbergen, University
and host cell targets for innovative anti-infective strategies.
of Ulm; Tobias Pukrop, University of Göttingen; Johan van Es, Hu-
selected puBlications
Rosenkrands, Statens Serum Institut, Copenhagen, DK.
brecht Institute, Utrecht, NL; Alun Kirby, University of York, UK; Ida
most important findings
lung, induced Wnt/b-Catenin signaling in murine macrophages,
Schaale K, Neumann J, Schneider D, Ehlers S, Reiling N.
Tuberculosis as a chronic inflammatory disease: Exploring
which was inhibited in the presence of a soluble Fzd1/Fc fusion
Wnt signaling in macrophages: Augmenting and inhibiting
the „novel“ functional role of WNT/Frizzled signaling in my-
protein. This pathway suppresses the activity of GSK3beta, a
mycobacteria-induced inflammatory responses. EUROPEAN
cobacterial infections
well known regulator of NF-kappaB-dependent gene transcrip-
JOURNAL OF CELL BIOLOGY 2010 Dec [Epub ahead of print]
Wnt proteins are secreted palmitoylated glycoproteins with
tion. This is well inline with the observation that Wnt3a reduced
doi:10.1016/j.ejcb.2010.11.004.
multiple functions in cell proliferation and migration as well
TNF release, suggesting that Wnt3a promotes anti-inflammatory
as tissue organization. They are best known for their role in
functions in murine macrophages. Our findings suggest that the
Neumann J, Schaale K, Farhat K, Endermann T, Ulmer AJ,
embryonic development and tissue homeostasis. In the last
evolutionarily conserved Wnt/β-catenin signaling is involved in
Ehlers S, Reiling N. Frizzled1 is a marker of inflammatory
years, Wnt signaling was also shown to be involved in the
balancing the inflammatory response to microbial stimulation of
macrophages, and its ligand Wnt3a is involved in reprogram-
regulation of inflammatory processes. In 2006 we were the
innate immune cells of vertebrate origin.
ming Mycobacterium tuberculosis-infected macrophages. FASEB
first who described an upregulation of Wnt proteins in a TLR/
JOURNAL 2010 Nov;24(11):4599-4612.
Figure 1:
Model of the
interplay of
TLR-,Wnt3a-,and
Wnt5a-induced
signal transduction
pathways in
macrophages.
(From Schaale
K, et al./Eur J
NF-κB dependent fashion by M. tuberculosis and other my-
Structural and functional characterisation of the intracellu-
cobacteria and identified a pro-inflammatory role for Wnt5a
lar habitat of pathogenic microorganisms: A unique, rapid
Klug K, Ehlers S, Uhlig S, Reiling N. Mitogen-activated pro-
and Fzd5 in human immune cells. It has now become clear
access to native M. tuberculosis phagosomes from primary
tein kinases p38 and ERK1/2 regulated control of Mycobacte-
that Wnt5a is not only induced by mycobacteria but is in-
cells
rium avium replication in primary murine macrophages is in-
Figure 2:
duced also in other infectious and inflammatory diseases
Some of the most vicious pathogens, among them Mycobacte-
dependent of tumor necrosis factor-alpha and interleukin-10.
Electron micro-
such sepsis, psoriasis, rheumatoid arthritis and atherosclerosis.
rium tuberculosis, reside in intracellular compartments, which
INNATE IMMUNITY Aug 2010 [Epub ahead of print] doi:
graph: Isolated
This prompted us to perform a comprehensive analysis of Wnt
are highly diverse and dynamic vacuoles. The detailed mo-
10.1177/1753425910377799
phagosome
signaling in experimental murine M. tuberculosis infection,
lecular composition of many of these vacuoles has remained
Cell Biol/2010)
containing
which is an well established model for a chronic inflammatory
insufficiently defined due to technical limitations in accessing
Schromm AB, Reiling N, Howe J, Wiesmuller K, Roessle M,
disease: We observed a reciprocal regulation of the Toll like
these compartments in a purified form. We have developed
Brandenburg K. Influence of serum on the immune recogni-
Receptor (TLR)/ Nuclear Factor kappa B (NF-κB) and the Wnt/β-
a novel, rapid and versatile method for the isolation of patho-
tion of a synthetic lipopeptide mimetic of the 19-kDa lipopro-
Catenin pathway after aerosol infection of mice with Mycobac-
gen-containing phagosomes from primary macrophages. We
tein from Mycobacterium tuberculosis. INNATE IMMUNITY 2010
The research of the group has been funded by grants of the
terium (M.) tuberculosis: Whereas pro-inflammatory mediators
developed a lipid-based procedure to label bacterial surfaces
Aug;16(4):213-25.
Deutsche Forschungsmeinschaft (SFB415 (Project C7) and DFG
were substantially increased, β-Catenin signaling was signifi-
and engineered a rapid immunomagnetic technique to isolate
cantly reduced. A systematic screen of Fzd homologs in infected
intact bacteria-containing compartments in less than two hours.
M. avium.
Grant support
Re1228/4-1) and the Bundesministerium für Bildung und ForscKrümmel B, Strassburg A, Ernst M, Reiling N, Eker B, Rath
hung (BMBF) Grant 01KI0784, WP D: “Pathogenic variability of
mice identified Fzd1 mRNA to be significantly upregulated dur-
Using magnetically labeled mycobacteria (M. tuberculosis, M.
H, Hoerster R, Wappler W, Glaewe A, Schoellhorn V, Sotgiu
clinical M. tb isolates: implications for the modulation of the
ing the course of infection. In vitro infection of murine macro-
avium) and Gram-positive bacteria (Listeria monocytogenes),
G, Lange C. Potential Role for IL-2 ELISpot in Differentiating Re-
immune system, infectivity and disease progression”.
phages led to a strong induction of Fzd1, which was dependent
we observed that microbial proteins are highly enriched in the
cent and Remote Infection in Tuberculosis Contact Tracing. PLOS
on TLRs, the myeloid differentiation response gene 88 (MyD88)
isolated phagosome containing fraction. A substantial change
ONE 2010 Jul;5(7):e11670.
and a functional NF-κB pathway. Flow cytometry demonstrated
in the molecular composition of the bacteria-containing phago-
an elevated Fzd1 expression on macrophages in response
somes over time documents pathogen- and host cell-specific
internal and external collaborations
to M. tuberculosis, which was synergistically enhanced in the
dynamics of intracellular trafficking and phagosome matura-
Members of the newly founded TB Center Borstel (TBCB) (Ste-
presence of IFN-g. Addition of the Fzd1 ligand Wnt3a, which
tion processes after infection. Mass spectrometric analysis of
fan Ehlers, Thomas Gutsmann, Christoph Hölscher, Otto Holst,
is constitutively expressed by bronchial epithelial cells in the
M. tuberculosis-containing phagosomes followed by Western
Christoph Lange, Buko Lindner, Stefan Niemann, Sabine Rüsch-
37
Dept. Molecular Infection biology • Microbial Interface Biology
PD Dr. Norbert Reiling
Models of Inflammation
inflammation
Janin Braun • Anne-Kathrin Claes • Sandra Hannemann • Dorothee Schultz • Christina Trabandt
Salmonella
fibrosis
Valdez Y, Grassl GA, Guttman JA, Coburn BA, Vallance
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). Salmonella is one of the major agents causing gastro-intestinal infection and inflammation causing morbidity and mortality world wide.
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 bacterial-triggered infection, iii) the mechanisms that lead to the development, propagation and resolution of intestinal inflammation and fibrosis. Finally we try to understand the similarities and differences in gut and lung inflammation and fibrosis.
most important findings
Role of NOD2 in intestinal inflammation: NOD-like receptors
(NLRs) are pattern recognition receptors for bacterial ligands.
blasts (Figure 1). This model is now being used to investigate
bacteria-epithelial interactions and crosstalk between epithelial cells and fibroblasts.
BA, Finlay BB. Nramp1 drives an accelerated inflammatory
response during Salmonella-induced colitis in mice. Cell Microbiol. 2009;11(2):351-62
internal and external collaborations
INTERNAL: Norbert Reiling, Division of Molecular Infection Biology; Ulrich Zähringer, Division of Molecular Infection Biology,
Helmut Haas, Division of Pneumology
EXTERNAL: John Baines, MPI Plön & Institute for Experimental
Medicine CAU Kiel; Dieter Kabelitz, Department of Immunolgy,
CAU Kiel; Ehrhardt Proksch and Jürgen Harder, Department of
Dermatology, CAU Kiel; Philip Rosenstiel, Institute for Clinical
and Molecular Biology, CAU Kie; Jürgen Brinckmann, Department of Dermatology, University of Lübeck; Dan Littman, Skirball
Institute, New York, USA; Brett Finlay, Michael Smith Laborato-
The NLR NOD2 is a cytoplasmic receptor for muramyl-dipep-
ries, UBC Vancouver, Canada; Alain Vandewalle, INSERM Paris,
tide (a part of the bacterial cell wall). Mutations of the NOD-
France
like receptor NOD2 are important risk factors for developing
Crohn’s disease, characterised by chronic intestinal inflamma-
Grant support
tion. It is believed that these mutations of NOD2 lead to a
Excellence Cluster “Inflammation at Interfaces”: Mouse models
loss-of-function phenotype of the receptor. However, how these
of inflammation
mutations result in un-controlled inflammation in the gut is not
Intramural grant CAU Kiel: “Mechanisms of collagen production
well understood. Using NOD2-/- and NOD2+/+ control mice, we
in fibrosis”
showed that the response to wildtype Salmonella was comparable. In contrast, infection with an attenuated Salmonella
mutant which produces an altered lipopolysaccharide (Salmo-
nella Typhimurium DmsbB) led to an increased inflammation
in NOD2-/- mice compared to controls. The LPS of Salmonella
Typhimurium DmsbB cannot trigger signalling through Toll-like
receptor (TLR4) indicating the importance of the crosstalk between TLR4 and NOD2 signalling. The mechanisms of this crosstalk are currently under investigation.
In vitro crypt organoid model: in order to investigate the
direct interaction of Salmonella Typhimurium with primary
epithelial cells and fibroblasts, we established and in vitro
epithelial crypt organoid model. Mouse intestine is minced
and embedded in a gelatin-matrix with the addition of growth
factors. Most cells die off, but intestinal epithelial stem cells
(found at the base of intestinal crypts) grow and differentiate
into all epithelial cell types of the gut including absorptive
enterocytes, goblet cells, paneth cells and enteroendocrine
cells. The crypt organoid is surrounded by sub-epithelial fibro-
Figure 1: H&E staining of colon crypt organoid grown from isolated
mouse intestine. F, fibroblasts; E, epithelial cells; G, goblet cells.
selected puBlications
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.
2010 Nov;12(11):1562-75. doi: 10.1111/j.1462-5822.2010.01488.x.
PubMed PMID: 20497179.
Kum WWS, Lee S, Grassl GA, Bidshahri R, Hsu K, Ziltener
HJ, Finlay BB. Lack of Functional P-Selectin Ligand Exacerbates Salmonella Serovar Typhimurium Infection. J Immunol. 2009;
182(10):6550-61
39
Dept. Molecular Infection biology • Models of Inflammation
Prof. Dr. Guntram Grassl
Crohn’s disease
infection
Immunobiology
Mast cell biology
proteases
Dr. Rajia Bahri • Katharina Biethahn • Anne-Katrin Brückner • Dr. Orietta D’Orlando • Hanno Ewers •
Dr. Nestor Gonzalez-Roldan • Diego Goyeneche-Patiño • Martina Hein • Manuel Hein • Andre Jenckel •
Frauke Koops • Rabea Langenhaun • Dr. Farhad Mirghomizadeh • Dr. Zane Orinska • Dr. Julia Polanski
Gesine Rode • Katrin Westphal
cytokines
allergy
MISSION
selected puBlications
Peter F. Johnson, NCI, Addgene, Rockville, USA
Over the last few years the Labgroup Immunobiology has developed a keen interest, namely to investigate the role, mechanism of action, survival and clinical relevance of mast cells in immunity.
Orinska Z, Föger N, Huber M, Marschall J, Mirghomizadeh
Ignazio Melero, Center for applied medical research, University
F, Du X, Scheller M, Rosenstiel P, Goldmann T, Bollinger
of Navarra, Spain
A, Beutler BA, Bulfone-Paus S. I787 provides signals for c-Kit
Laurence Zitvogel, Institut Gustave Roussy, Villejuif, France
most important findings
N-nitrosurea (ENU) mutagenesis in C57BL/6J mice – an A to
receptor internalization and functionality that control mast cell
Elena Voronov, Institut of Immunology, Beer Sheva, Israel
Novel aspects of mast cell biology
T transversion at position 2388 (exon 17) of the c-Kit gene,
survival and development. Blood. 116:2665-75, 2010
Axel Roers, Institut für Immunologie, Universität Dresden
Recently our group could demonstrate that the activity of mast
resulting in the isoleucine 787 substitution by phenylalanine
cell proteases is essential in the control of bacterial clear-
(I787F) has profound consequences on mast cell develop-
Stelekati E, Bahri R, D‘Orlando O, Orinska Z, Mittrücker
ance in vivo and that the expression of these mast cell pro-
ment and signaling. The mice carrying the described muta-
HW, Langenhaun R, Glatzel M, Bollinger A, Paus R, Bulfo-
Gillian Griffiths, Cambridge Institute for Medical Research, Ad-
teases is under the control of intracellularly stored cytokines
tion lack mucosal and connective tissue-type mast cells. Fur-
ne-Paus S. Mast cell-mediated antigen presentation regulates
denbrooke’s Hospital, England
(Orinska et al., Nature Medicine 2007). Furthermore we could
thermore, bone marrow derived mast cells differentiated in
CD8+ T cell effector functions. Immunity. 31:665-76, 2009
show that in addition to their well known effector functions
vitro are strongly impaired in SCF-induced cytokine produc-
Marina Scheller, Max-Delbrueck-Center for Molecular Med.
Tumorigenesis and Cell Differentiation
Erietta Stelekati, Penn Institute of Immunology, Pennsylvania,
USA
in allergy, mast cells profoundly regulate CD8 T cell immu-
tion, degranulation and survival. The c-Kit downstream sig-
Mirghomizadeh F, Bullwinkel J, Orinska Z, Janssen O, Pe-
Andreas Radbruch, Hyu-Dong Chang, Deutsches Rheumafor-
nity in an antigen dependent-manner and thereby modulate
naling in mast cells carrying the I787F mutation is normally
tersen A, Singh PB, Bulfone-Paus S. Transcriptional regulation
schungszentrum, Berlin
important adaptive immune responses, e.g. in autoimmune
initiated but is not sustained at later time points. In addition,
of mouse mast cell protease-2 by interleukin-15. J Biol Chem.
Dunja Bruder, Helmholtz Center for Infection Research
diseases and viral infection (Stelekati et al., Immunity 2009).
this mutation influences c-Kit receptor ubiquitination and af-
284:32635-41, 2009
Grant support
fects c-Kit internalization. These studies underline the concept
Control of mast cell development, survival and function
that sustained c-Kit signaling and receptor internalization are
Mirghomizadeh F, Winoto-Morbach S, Orinska Z, Lee KH,
SPP 1394 DFG/OR 101/2-1 (2009 – 2012)
by c-Kit I787
required for mast cell development, activation and survival
Schütze S, Bulfone-Paus S. Intracellular IL-15 controls mast cell
Mast cells as linkers of antiviral defence and autoimmunity
Mast cell differentiation, survival and effector functions are
(Orinska et al., Blood 2010).
survival. Exp Cell Res. 315:3064-75, 2009
DFG/SFB 415/A10
regulated by the membrane tyrosine kinase c-Kit upon interaction with the ligand stem cell factor (SCF). We have recently
Regulation of mast cell function by the actin cytoskeleton
internal and external collaborations
Identifizierung und Charakterisierung neuer Interaktionspartner
described that a single point mutation induced by N-ethyl-
Mast cell activation causes immediate degranulation and
Regional collaborations:
in der Signaltransduktion durch den IL-15 Rezeptor
release of pro-inflammatory mediators in a process that in-
University of Lübeck: Thomas Bollinger, Institut für Mikrobiologie
Kit787F mice
volves the reorganization of the actin cytoskeleton. The regu-
und Hygiene
DFG/TR-SFB22/A14
carrying a point
latory pathways and functional links between mast cell activa-
Peter König, Institut für Anatomie
Die Rolle von CD8+ regulatorischen T-Zellen und Mastzellen
mutation in
tion, cytoskeletal reorganization and mast cell degranulation
Jörg Köhl, Institut für Systemische Entündungsforschung
bei der Induktion und Chronifizierung von allergischem Asthma
c-Kit are mast
are, however, only insufficiently characterized. Thus, we have
University of Kiel: Cooperations within SFB 415: Dieter Adam,
cell deficient.
initiated a project to further define the role of actin cytoskel-
Paul Saftig, Ottmar Janßen, Stefan Schütze, Philip Rosenstiel
Cytospins from
etal dynamics on mast cell function. Utilizing genetic mouse
Figure 1:
FP7-Health-2007-A 201461
European initiative to improve knowledge, treatment and sur-
wild type or
models, our data indicate a critical role of the actin regulato-
National and international collaborations:
Kit787F mice
ry proteins Coronin1a and Coronin1b on exocytic pathways in
Tiziana Musso, University of Torino
were subjected
mast cells. In a multidisciplinary approach that combines bio-
Hans-Willi Mittrücker, University of Hamburg
FP7-Health-2007-B223151 (2009 – 2012)
to toluidine blue
chemical, cell biological and mutational studies we currently
Oliver Pabst, University of Hanover
Understanding inflammation-associated tumorigenesis for the
staining. Mast
analyze the stimulus dependent regulation of coronin proteins
Friedrich Nolte, Institut für Immunologie, University of Hamburg
rational design of novel anti cancer therapeutic strategies
cells are dark
in mast cells and characterize molecular requirements for
Michael Huber, Universitätsklinikum Aachen
(INFLA-CARE)
blue stained
the regulatory function of coronin on cellular degranulation.
Klaus Ruckdeschel/Martin Äpfelbacher, University of Hamburg
cells. Photos
These mechanistic in vitro studies are complemented by an
Inger Gjertsson, University of Gothenburg, Sweden
were taken
in vivo analysis of the physiological role of coronin proteins in
Udo zur Stadt, University of Hamburg
with a light
anaphylactic reactions. As cells likely utilize common regula-
Peter Katsikis, Dept. of Microbiology and Immunology, Drexel
microscope
tory themes, such as the coronin/actin cytoskeletal pathway to
University, Philadelphia, USA
under 400x
control exocytic processes, our findings will likely also apply
Takashi Fujita, Institute for Virus Research, Kyoto University, Japan
magnification.
to other secretory cells of the immune system.
Martin Turner, Babraham Research Center, Cambridge, England
vival of haemophagocytic syndromes in children (CureHLH) 41
Dept. immunology and cell Biology • Immunobiology
Prof. Silvia Bulfone-Paus
Molecular Immunology
Inflammatory
signal transduction
W. Cheng • Dr. Gulnar Fattakova • Azhar Kamal • Lekha Nath • Xuan Hung Nguyen • Patricia Prilla •
Beate Schurek
Immune disorders
Kyeong-Hee Lee, PhD
and LYST are preferentially expressed in hematopoietic tissues
internal and external collaborations
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 autoimmunity and allergy/asthma, as well as determining perspectives for clinical interventions.
and are particularly highly expressed in mast cells.
• Dr. Tak Mak, Campbell Family Institute for Breast Cancer
Our ongoing studies implicate RIN3 and LYST in mast cell
Research, Toronto, Canada
function: preliminary data suggest that RIN3 acts as a negative
• Prof. Dr. Dieter Adam, University of Kiel, Germany
regulator, while LYST exhibits a positive regulatory function on
• Dr. Karl Lang, Humboldt Reserch Group, University of
mast cell degranulation.
Düsseldorf, Germany
most important findings
In addition, we could demonstrate that human Toso-specific
Project I: Characterization of novel regulatory mechanisms
monoclonal antibodies significantly enhance the sensitivity to
Since RIN3 is involved in regulating membrane dynamics at
of death receptor signaling pathways leading to the
death receptor-mediated apoptosis, and block TNFα-mediated
early endosomes, while LYST acts at the level of lysosomes,
regulation of inflammation and autoimmunity
pro-inflammatory signaling pathways. Thus, Toso specific
our results on the opposing functions of RIN3 and LYST for mast
Members of the tumor necrosis factor receptor (TNFR)
blocking antibodies may open up therapeutic prospects for
cell degranulation may have implications for our understanding
superfamily, Fas (CD95) and TNFR1 (CD120a) are critically
the treatment of immunopathological disorders associated
on how and to what extent different endosomal compartments
involved in the regulation of cellular apoptosis and survival
with TNFα-induced cytotoxicity. As Toso overexpression is often
contribute to mast cell function. Hence these studies may help
in the immune system. A marked functional dichotomy of
observed in autoimmune T cells and leukemias, results from our
to elucidate the mechanisms leading to mast cell mediated
these receptors is that they can transmit both, death inducing,
antibody blocking studies could also be highly relevant for the
inflammatory disease.
as well as activating signals through the same receptor in
development of novel clinical applications for the treatment of
response to the same ligand. Our studies have improved our
immune disorders and hematological neoplasms.
• Prof. Dr. Marcus E. Peter, The Ben May Institute for Cancer
Research, Chicago, USA
• Dr. Andrew C. Chan, Vice President Research Immunology,
Genentech Inc., USA
• Asthma/Allergy Group in the Immunology Department at
Genentech Inc. USA
selected puBlications
Xuan-Hung Nguyen, Philipp A. Lang, Karl S. Lang, Gulnar
understanding of how these different types of TNF receptor
signaling (activation vs. apoptotic signaling) are regulated
Project II: Integrating molecular and cellular aspects of
Fattakhova, Dieter Adam, Niko Föger, Md. Azhar Kamal,
by the formation of specific receptor complexes in distinct
membrane trafficking and inflammation:
Patricia Prilla, Sabine Mathieu, Christina Wagner, Tak Mak,
subcellular compartments (published at FEBSJ. 2009, EMBO J.
The role of the membrane trafficking regulators RIN3 and
Andrew C. Chan and Kyeong-Hee Lee. Toso regulates the
2006). These findings provide new insights into how membrane
Lyst in mast cell function
balance between apoptotic and non-apoptotic death receptor
trafficking and the sub-cellular localization of cellular proteins
Mast cells play a pivotal role in human allergic and
signaling by facilitating RIP1 ubiquitination
are integrated into death receptor-mediated apoptotic
inflammatory disease. Mast cell activation leads to immediate
signaling pathways.
release of mediators from secretory granules in a process
(in revision)
commonly known as degranulation. This mast cell degranulation
Karl S. Lang, Philipp A. Lang, Andreas Meryk, Louis-Martin
Figure 1: Nuclearcyto-plasmic redistrubution of FADD following CD95L
We have also recently identified and characterized a novel
intensively requires specifically regulated vesicle transport
Boucher, Jillian Haight, Pauline Funkner, Andrew Wakeham,
stimulation. hCD95 expressing A20 cells were stimulated for 0 or 30 min
regulator of Fas and TNFR signaling, named Toso (Faim3), which
and membrane fusion processes. However, the molecular
Nadine Honke, Mike Recher, Alexander A. Navarini, Nadine
with mAb against hCD95. Cells were fixed and stained for FADD (green)
has been implicated in human autoimmune and neoplastic
mechanisms responsible for the intracellular trafficking of
Honke, Pamela S. Ohashi, Dieter Häussinger, Kyeong-Hee
and the early endosome marker EEA-1 (red).Taken from FEBS J, 2009 vol.
disorders. In our studies, we could provide clear genetic in
vesicles and regulated fusion of mast cell granules with the
Lee* and Tak W. Mak* (* shared senior authorship).
276 (15) cover picture
vivo evidence that the transmembrane protein Toso regulates
plasma membrane are still largely unknown.
Toso-deficient granulocytes are pre-primed but show impaired
Grant support
effector function
death receptor induced pro-survival vs. pro-apoptotic signaling
in immune cells. In response to death receptor stimulation, Toso
We here investigate the role of two distinct regulators of
facilitates the ubiquitination of RIP1 and mediates the activation
membrane trafficking, RIN3 and LYST, during mast cell
(in revision)
1. Marie Curie International Reintegration Grant: FP7-PEOPLE2007-4-3-IRG
of MAPK and NF-κB signaling cascades. Our findings provide
degranulation. RIN3 is a specific guanine nucleotide exchange
Niko Föger, Silvia Bulfone-Paus, Andrew C. Chan and
2. Fritz Thyssen Research Foundation: Az. 10.08.2.155
strong evidence for non-apoptotic signaling of death receptors
factor (GEF) for the small GTPase Rab5, which is involved
Kyeong-Hee Lee. Subcellular compartmentalization of FADD as
3. German Research Foundation (DFG): LE 1254/2-1
and provide a new molecular basis for how the execution
in the regulation of endocytosis and endosome-mediated
a novel level of regulation in death receptor signalling FEBS J
4. Cluster of Excellence in Inflammation Research
of death receptor mediated pro-survival and pro-apoptotic
intracellular vesicle trafficking. LYST is a lysosomal trafficking
cover story; 2009, August, 276 (15):4256-65
signaling can be regulated in the immune system.
regulator and mutations in the LYST gene cause the Chediak–
Higashi syndrome (CHS) in humans, which is characterized by
Mirghomizadeh F, Winoto-Morbach S, Orinska Z, Lee K. H.,
physiologic
severe immunodeficiency and recurrent bacterial infections. A
Schütze S, Bulfone-Paus S. Intracellular IL-15 controls mast cell
demonstration of the in vivo importance of Toso by demonstrating
prominent feature of this disease is also the accumulation of
survival. Exp Cell Res. 2009 Oct 15;315(17):3064-75
that Toso is critically involved in TNFα-mediated liver injury.
enlarged lysosome-related granules in a variety of cells. RIN3
Furthermore,
our
studies
provide
the
first
- Knockout mouse generation for the study of inflammatory
disease models
- PhD student Scholarship for Inflammation Research
43
Dept. immunology and cell Biology • Molecular Immunology
MISSION
Biochemical Immunology
Mast cell-neutrophil interaction
asthma
Cindy Hass • Diana Heinrich • Dr. Brigitte Kasper • Anika Kasprick • Lisette Leonhardt • Carola Schneider
Dr. Xinhua Yu
autoimmunity
tissue damage
MISSION
process. CXCL4-treated monocytes significantly upregulate sphin-
tian-Albrechts University of Kiel. Animal models of autoimmune
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.
gosine kinase 1 (SphK1) mRNA and CXCL4 induces SphK1 en-
dermatoses - D. Zillikens, R. Ludwig, Institute of Dermatology,
zyme activity as well as its translocation to the cell membrane.
University of Lübeck.
Furthermore, we could show that pharmacological inhibition of
SphK results in reversal of CXCL4-induced monocyte survival, cy-
Figure 1:
tokine expression, and release of oxygen radicals, which was
Neutrophil-deri-
most important findings
against type VII collagen. In our study we analyze the role of
confirmed by the use of SphK1-specific small interfering RNA.
ved antimicro-
Allergic asthma appears as a chronic eosinophilic inflammation
MC and their products in EBA and evaluate whether these cells
Furthermore, CXCL4-mediated respiratory burst is dependent on
bial peptides
of the airways which was, until recently, seen as directly causative
could serve as potential therapeutic targets for the treatment of
a very rapid activation of PI3K, Syk and p38 MAPK, while rescue
HNP and
for the asthma pathology in terms of airway hyperreactivity and
this disease.
from apoptosis, which is accompanied by inhibition of caspases,
LL-37 induce
is controlled by SphK1 and its downstream element Erk (Fig. 2).
degranulation
remodelling. However, disappointing results from anti-IL-5 as wells
as IL-12 studies revealed that the accumulation of eosinophils in
To determine whether MC play an essential role in EBA in gen-
Taken together, these data assign SphK1 as a central regulator
in human lung
asthma is not directly related to airway dysfunction. In our study
eral, we passively induced EBA in wildtype as well as mast cell-
of acute and delayed monocyte activation and suggest SphK1
mast cells.
we investigated the pathophysiological role of neutrophils in the
deficient mice (C57BL/6J KitW-sh/KitW-sh) by administration of rabbit
as a potential therapeutic target to suppress pro-inflammatory
responses induced by CXCL4.
disease. By using an acute model of allergic asthma in vivo, we
IgG against mouse collagen 7 (amcol7). In contrast to our hy-
could show that depletion of neutrophils is accompanied by a
pothesis, mice of the MC-deficient strain developed a significant
reduced eosinophilia in the BAL and less inflammatory cellular
severe phenotype of experimental EBA as compared to the cor-
selected puBlications
infiltrates in the lung tissue. Furthermore, goblet cell hyperplasia
responding wildtype controls. However, these results could not be
Schiemann F, Brandt E, Gross R, Lindner B, Mittelstadt J,
and airway hyperreactivity were significantly decreased in the
confirmed in a second strain of mast cell-deficient mice (WBB6F1
Sommerhoff CP, Schulmistrat J, Petersen F. The Cathelicidin
latter animals. Our findings provide first evidence that neutrophils
KitW/KitW-v), where no significant difference between knockout
LL-37 Activates Human Mast Cells and Is Degraded by Mast
are not only critically involved in allergic airway inflammation
and the corresponding littermates in the induction of the disease
Cell Tryptase: Counter-Regulation by CXCL4. JOURNAL OF IM-
but also in the development of the asthma pathology in terms of
could be observed. Since deficiency of c-Kit affects beside mast
MUNOLOGY. 2009; 183:2223-2231.
mucus production and airway hyperreactivity.
cells also other cells of the immune system, we decided to use for
The question whether mast cells could be involved in the recruita series of experiments in vitro using human primary lung mast
Figure 2:
future experiments the transgenic Mcpt5-Cre iDTR double positive
Kasper B, Winoto-Morbach S, Mittelstädt J, Brandt E, Schü-
Sphingosine ki-
mice, a model of a c-Kit independent mast cell ablation.
tze S, Petersen F. CXCL4-induced monocyte survival, cytokine
nase 1 (SphK1)
expression, and oxygen radical formation is regulated by sphin-
mediates
This new model will allow an investigation of mast cell functions
gosine kinase 1. EUROPEAN JOURNAL OF IMMUNOLOGY.
immediate
2010; 40:1162-1173.
and delayed
ment of neutrophils into the allergic lung was addressed first by
cells. We could should that FcεRI-activated mast cells release me-
without defects in other cell lineages and thus ensuring a proper
diators chemotactic for neutrophils. These mediators are under
immune response in the passive model of EBA. In Bullous pem-
current investigation and at least one neutrophil-active chemotax-
phigoid (BP), an EBA-related autoimmune bullous disease associ-
Yu X, Holdorf K, Kasper B, Zillikens D, Ludwig RJ, Petersen
CXCL4-acti-
in could be identified so far. Most intriguingly, mast cell-neutrophil
ated with autoantibodies to the hemidesmosomal proteins BP230
F. FcgammaRIIA and FcgammaRIIIB Are Required for Autoanti-
vated human
monocytes.
signaling in
interaction appears to be a bidirectional process. We could show
and BP180 (collagen 17), we have investigated the contribution
body-Induced Tissue Damage in Experimental Human Models
that the neutrophil-derived antimicrobial peptides LL37 and HNP
of individual Fcg receptors on human neutrophils in the pathogen-
of Bullous Pemphigoid. JOURNAL OF INVESTIGATIVE DERMA-
induce degranulation in purified human lung MC (Fig. 1). Intrigu-
esis of the disease. By using an ex vivo and a newly established
TOLOGY. 2010; 130:2841-2844.
ingly, as a consequence LL37 rapidly undergoes limited cleavage
in vitro model of BP we could demonstrate that both, FcgRII and
by a released mast cell tryptase resulting in the inactivation of
FcgRIII, are essential for dermal-epidermal separation in the dis-
internal and external collaborations
projekt A11
the peptide. Based on our findings in vitro, we presently analyze
ease.
Mast cell-neutrophil interaction in asthma - E. Brandt, Z. Orin-
DFG SFB 415 “Spezifität und Pathophysiologie von Signaltrans-
ska, O. Holst, B. Lindner, Research Center Borstel; H. Renz, H.
duktionswegen”, Teilprojekt B6
CXC chemokine ligand 4 (CXCL4) belongs to the CXC-chemo-
Garn, L. Marsh, University of Marburg; R. Hoffmann, O. Prazeres
Cluster of Excellence „Inflammation at Interfaces”; Research
kine family and represents a broad activator of acute as well
da Costa, Technical University of Munich; P. König, Institute of
Area H: IRN Autoimmunity to Type VII Collagen
Epidermolysis bullosa acquisita (EBA) is an acquired chronic
as delayed functions in monocytes. By analyzing signaling path-
Anatomy, University of Lübeck. S100A8/A9 proteins in asthma
SPP AI “Mechanismen und innovative Modulation von organ-
subepidermal blistering disease of skin and mucous membranes
ways controlling CXCL4-mediated monocyte functions, we could
and autoimmunity - T. Vogl, University of Münster. Analysis of
spezifischer Autoimmunität”, Teilprojekt B4
characterised by circulating and tissue-bound autoantibodies
define an essential role for sphingosine kinase 1 (SphK1) in this
spingosine kinases - S. Schütze, Institute of Immunology, Chris-
DFG PE 967/1-1
neutrophil infiltration into the lungs of mast cell deficient KitW-sh
mice in acute allergic asthma.
45
Grant support
DFG SFB/TR 22 “Allergische Immunantworten der Lunge“, Teil-
Dept. immunology and cell Biology • Biochemical Immunology
Prof. Dr. Frank Petersen
Biological Chemistry
inflammation
allergy
Christine Engellenner • Dr. Oranos Ghulam • Gabriele HuSS • Dr. Florian Schiemann • Jan Schulmistrat
neutrophils
mast cells
platelet chemokines
MISSION
of the vascular wall, molecules from out the blood and even
human macrophage infection with HIV-1 and potentiates virus
To investigate the roles of chemokines and proteases in the communication of platelets, neutrophils and mast cells during allergic and non-allergic inflammation
blood platelets may invade the surrounding tissue. Previous
replication. Innate Immun 2009; 15: 368-379.
findings in our laboratory, demonstrating that the plateletetderived chemokine CXCL4 may act as an inhibitor of major
Kasper B, Winoto-Morbach S, Mittelstädt J, Brandt E, Schüt-
most important findings
cells (Fig.1). Moreover, treatment with PAR1-AP and PAR4-AP
mast cell proteases chymase and tryptase are in line with this
ze S, Petersen F. CXC chemokine ligand 4 (CXCL4)-induced
Expression and regulatory function of proteinase-activated
stimulated degranulation in these cells.
hypothesis.
monocyte survival, cytokine expression, and oxygen radical
receptors on human mast cells
4. Natural ligands of PAR1 and PAR4 (thrombin) and PAR2
formation is regulated by sphingosine kinase 1. Eur J Immunol
Proteinase-acitvated receptors (PARs) form a small group of
(trypsin) also induced intracellular Ca-ion transients, but sur-
Figure 1:
related proteins (PAR1 to 4) within the large familiy of G protein-
prisingly were unable to challenge a degranulation respon-
Induction of
coupled seven transmembrane-domain receptors. Their mecha-
se. The latter result was quite unexpected, since in murine
intracellular Ca-
siology and therapeutic aspects. Cell Molec Life Sci 2010; 67:
2363-2386.
2010; 40: 1-12.
Flad H-D, Brandt E. Platelet-derived chemokines: pathophy-
nism of activation is unique, in that their extracellular N-terminal
mast cells proteinases thrombin and trypsin were shown
ion transients
stretches of sequence are susceptible to proteolytic truncation
to induce degranulation. We therefore further examined in
in human
at specific sites, resulting in the exposure of a tethered ligand
lung mast cells, whether natural PAR ligands might modu-
primary lung
domain that binds to and activates the receptor. PARs have
late/enhance mast cell responses such as FcεRI-mediated
mast cells by
degranulation.
PAR-activating
mokine ligand 4 (CXCL4) downregulates CC chemokine recep-
been found to be of widespread physiological importance
Schwartzkopff F, Petersen F,Grimm TA, Brandt E. CXC che-
by participating in a number of fundamental processes such
5. Unexpectedly thrombin did not enhance but inhibited FcεRI-
peptides (A),
tor expression on human monocytes.
as hemostasis, tissue repair and inflammation by reacting to
mediated mast cell degranulation. Inhibition with a physio-
and thrombin
Innate Immun 2010 [Epub ahead of print]
cellular or tissue-derived proteases. Since their discovery on
logical dosage of thrombin was most effective upon brief
(B).
blood platelets, PARs have been detected on many tissue and
6. The inhibitory effect by thrombin is dependent on the cata-
cells, neurons, cells of the gastrointestinal tract and mast cells.
lytic activity of the proteinase, as seen upon application of
Recent evidence obtained in mice suggests that PAR expressi-
thrombin-directed inhibitors.
on in mast cells could be relevant in allergic reactions. As cor-
7. Inhibiton by thrombin appears to be specific for FcεRI-
responding studies on mast cells of human origin are still rare
mediated degranulation, as no effect was seen with res-
and predominantly rely on experiments performed with mast
ponses to several other known inducers of the mast cell
cell lines, we have analysed the expression and functionality of
response.
PARs on primary mast cells isolated from human lung and skin.
internal and external collaborations
preincubation of the cells with the protease (Fig.2).
immune cells, including leukocytes, endothelium, vascular wall
8. To assess the role of PARs in thrombin-mediated inhibition
indirect and direct experimental approaches were used.
Our most important findings are the following:
Thus, the general ability of PAR1 and PAR4 to transduce
1. Human primary mast cells from lung and skin both express
inhibitory signals was evident from the potential of PAR1-
mRNA for PAR1, 2, 3, and 4. Expression for all four PARs
AP and PAR2-AP to downregulate mast cell degranulati-
is strong and reproducible in lung mast cells, while PAR2
on. Further analyses using intracellular receptor-specific
expression is variable in skin mast cells.
antagonists (pepducins) likewise revealed that inhibitory
2. Cell surface expression of receptor proteins was always
signal(s) mediated through PAR1 and PAR4 were involved,
detectable for PAR1, was variable for PAR4, while PAR2
and that both receptors acted in concert to mediate the
and PAR3 could not be detected in either type of mast cell.
thrombin effect.
Petersen F, Kasper B, Mittelstädt J, Gross R, Division of Biochemical Immunology, Research Center Borstel
Figure 2:
Effect of preincubation with
thrombin on
anti-IgE-induced degranulation (release of
β-hexosaminidase enzymatic
activity) of
primary human
lung mast cells.
selected puBlications
Schiemann F, Brandt E, Gross R, Schulmistrat J, Lindner B,
Instead, strong intracellular PAR2 protein expression was
Altogether our results suggest that in the human system cer-
Sommerhoff C, Petersen F. The cathelicidin LL-37 activates hu-
3. Nevertheless, evidence for the expression of functional
tain blood-borne and tissue-derived proteases may have a role
man mast cells and is degraded by mast cell tryptase: counter-
PAR1, 2, and 4 was obtained by stimulating lung and skin
in regulating mast cell activation through PARs. This may be
regulation by CXCL4. J Immunol 2009; 183: 2223-2231.
mast cells with receptor-specific synthetic peptides (PAR-
instrumental in limiting mast cell activation during the acute
AP), where PAR1-AP, PAR-2AP, and PAR4-AP reproducibly
response to allergens, where due to inflammation-associated
Schwartzkopff F, Grimm TA, Lankford CSR, Fields K, Wang
induced intracellular Ca-ion transients in both types of mast
liberation of vasoactive substances and increased leakyness
J, Brandt E, Clouse KA. CXCL4 (platelet factor 4) facilitates
seen in lung and skin mast cells.
47
Lindner B, Division of Immunochemistry, Research Center Borstel
Clouse KA, Center for Drug Evaluation and Research, US Food
and Drug Administration, Rockville, Maryland, USA
Sommerhoff C, Department of Clinical Chemistry and Clinical
Biochemistry, Ludwig-Maximilians-Universität, München
Grant support
Deutsche Forschungsgemeinschaft, SFB/TR22 „Allergische Immunantworten der Lunge“, Projekt A11.
Dept. immunology and cell Biology • Biological Chemistry
Prof. Dr. Ernst Brandt
VETERINARY INFECTION BIOLOGY AND IMMUNOLOGY
Vector-borne diseases
Jassim Abdo • Doreen Beyer • Birgit Kullmann • Junlong Liu • Monika Mackiewicz • Heike Müller • Bogdan Oltean • Britta Petersen • Dr. Stefanie Renneker • Prof. Dr. Ulrike Seitzer • Susan Tschitschmann
vaccine development
diagnostics
host-pathogen-interaction
theileria
MISSION
the microtubuli network of their host cell via the membrane protein
control of emerging viral vector-borne zoonotic diseases: Arbo-
The major interest of our research activities lies in the investigation of host-vector-pathogen interactions and is focused on three main areas. Firstly, we are engaged in the development of diagnostics and vaccines for the tick-borne parasitic diseases ovine and bovine theileriosis. A second focus of our research aims at the elucidation of the molecular mechanisms involved in the host-parasite interaction leading to the transformation by Theileria annulata schizonts of the host cells they infect. The third research area involves investigations on the hosts immune response to tick bite and on vector competence. Lastly, we are committed to the dissemination of knowledge and capacity building in emerging and developing countries.
TaSP. PARASITOLOGY RESEARCH. 2010 Apr; 106(5):1085-1102.
Zoonet. EUROSURVEILLANCE. 2009 Mar; 14(12):11-14.
Salih D, EL Hussein A, Ahmed J, Seitzer U. Comparison
Between Reverse Line Blot and Enzyme-linked Immunosorbent
Assay in Diagnosis of Major Tick-Borne Diseases of Cattle in
Southern Sudan. TRANSBOUNDARY AND EMERGING DISEASES.
2010 Apr; 57(1-2):61-62.
Müller H, Aysul N, Liu Z, Salih D, Karagenc T, Beyer D,
Figure 1:
Lateral flow
device for detection of Thei-
leria annulata
infection. Left:
negative serum
sample. Right:
most important findings
of isolated schizonts. Curing infected cells of the parasite
Kullmann B, Ahmed J, Seitzer U. Development of a Loop-
Several advancements in the establishment of diagnostic tools
resulted in a time dependent translocation of p53 into the
mediated Isothermal Amplification (LAMP) Assay for Rapid
for detection of piroplasmosis were made. Thus, two proteins
host cell nucleus and the up-regulation of the pro-apoptotic
Diagnosis of Babesia canis infections. TRANSBOUNDARY AND
of Theileria uilenbergi were identified and characterized (TuIP,
Bax and Apaf-1 and the down-regulation of the anti-apoptotic
EMERGING DISEASES. 2010 Apr; 57(1-2):63-65.
clone-9) against which antibodies are generated in the infected
Bcl-2 proteins. Subsequent experiments could show that
host. Both antigens were found suitable for the establishment
whereas specific killing of the parasite with buparvaquone
Liu Z, Wang Z, Yin H, Luo J, Zhang B, Kullmann B, Abdo J,
tion of the Thei-
and application of a recombinant protein based indirect
led to apoptosis of the host cell, inhibition of either p53 or
Salih D, Ahmed J, Seitzer U. Identification of Theileria uilenbergi
leria annulata
ELISA for detection of T. uilenbergi infection. In the area of
Bax significantly reduced buparvaquone-induced apoptosis of
immunodominant protein for development of an indirect ELISA
schizont TaSP
tropical theileriosis, an epidemilogical study was performed
the transformed cells. Thus, infection with Theileria annulata
for diagnosis of ovine theileriosis. INTERNATIONAL JOURNAL
protein with
in northern Iraq, showing a high prevalence of the disease.
does not induce the p53 apoptotic pathway of host cells by
FOR PARASITOLOGY. 2010 Apr; 40(5):591-598.
microtubules of
In addition, a previously established competitive ELISA was
a mechanism involving cytoplasmic sequestration of p53. The
validated. A major advancement was the establishment of
close association of host cell p53 with the parasite membrane
Ahmed J, Sparagano O, Seitzer U. One health, one medicine:
telophase.
an immunochromatographic strip test (lateral flow device)
implies that the parasite interacts either directly with p53 or
Tackling the challenge of emerging diseases. TRANSBOUNDARY
A) TaSP;
for rapid detection of infection with Theileria annulata. This
mediates cytoplasmic sequestration of p53 by interacting with
AND EMERGING DISEASES. 2010 Apr; 57(1-2):1-2.
B) tubulin;
point-of-care and easy to perform test may greatly advance
other host cell proteins regulating p53 localization.
positive serum
sample.
Figure 2:
Close associa-
a host cell in
C) DNA;
Haller D, Mackiewicz M, Gerber S, Beyer D, Kullmann B,
diagnostics for tropical theileriosis in the field (Fig.1). In the
D) overlay.
research area dealing with the molecular interplay of the
selected puBlications
transforming Theileria annulata schizont with its host cell, we
Müller A, Goldmann T, Seitzer U. Measuring Immune
of p53 promotes survival in leukocytes transformed by Theileria.
internal and external collaborations
could show that a membrane protein of the parasite (TaSP)
Responses In Situ: Immunofluorescent and Immunoenzymatic
ONCOGENE. 2010 Mar; 29(21):3079-3086.
Through the intensive participation in EU funded collaborative
interacts with the host cell microtubule network (Fig.2). Analysis
Techniques. METHODS IN MICROBIOLOGY. 2010; 37:421-437.
Schneider I, Ahmed JS, Seitzer U. Cytoplasmic sequestration
projects, DAAD (German Academic Exchange Service) funded
Al-Saeed A, Omer L, Abdo J, Habibi G, Salih D, Seitzer
Strategic Academic Partnerships, DFG (German Research
cell mitosis indicated that TaSP co-localizes and interacts
Abdo J, Kristersson T, Seitzer U, Renneker S, Merza M,
U, Ahmed J. Epidemiological studies on tropical theileriosis
Foundation) funded German-African Cooperation Projects in
with the spindle poles, the mitotic spindle apparatus and the
Ahmed J. Development and laboratory evaluation of a lateral
(Theileria annulata infection of cattle) in Kurdistan Region, Iraq.
Infectiology and the Leibniz Graduate School „Model Systems for
midbody. Moreover, TaSP was demonstrated to be localized
flow device (LFD) for the serodiagnosis of Theileria annulata
PARASITOLOGY RESEARCH. 2010 Jan; 106(2):403-407.
Infectious Diseases“ the Division of Veterinary Infection Biology
to the microtubule organizing center and to physically interact
infection. PARASITOLOGY RESEARCH. 2010 Oct; 107(5):1241-1248.
Renneker S, Abdo J, Ahmed J, Seitzer U. Field validation of a
the national and international level (Europe, Asia, Africa, Latin
America and Australia).
of the interaction of TaSP and host microtubuli during host
with gamma-tubulin. These data support the notion that the
and Immunology is linked to numerous research institutions at
TaSP – microtubule interaction may be playing a potential role
Abdo J, Liu Z, Yin H, Kullmann B, Ahmed JS, Seitzer
competitive ELISA for detection of Theileria annulata infection.
in parasite distribution into daughter host cells and gives rise
U. Identification of clone-9 antigenic protein of Theileria
PARASITOLOGY RESEARCH. 2009 Dec; 106(1):47-53.
to the speculation that TaSP may be involved in regulation
uilenbergi and evaluation of its application for serodiagnosis.
of microtubule assembly in the host cell. A further study
PARASITOLOGY RESEARCH. 2010 Aug; 107(3):517-524.
investigated the potential role of p53 in survival of the infected
49
Grant support
Ahmed J, Bouloy M, Ergonul O, Fooks A, Paweska J,
External funding new in 2009/2010: PiroVac, Postick (EU FP7),
Chevalier V, Drosten C, Moormann R, Tordo N, Vatansever
German-African Cooperation Projects in Infectiology (DFG), Leibniz
host cell. We found p53 to be largely localized in the host
Seitzer U, Gerber S, Beyer D, Dobschanski J, Kullmann B,
Z, Calistri P, Estrada-Pena A, Mirazimi A, Unger H, Yin H,
Graduate School, DAAD; running in 2009/2010: Arbo‑Zoonet (EU
cell cytoplasm and associated with the parasite membrane
Haller D, Ahmed J. Schizonts of Theileria annulata interact with
Seitzer U. International network for capacity building for the
FP7), ICTTD-3, ConFluTech, ASEMDialog (EU FP6).
Dept. immunology and cell Biology • VET. INFECTION BIOLOGY & IMMUNOLOGY
Prof. Dr. Jabbar S. Ahmed
Immune Cell-Analytics
immune cell regulation
Franziska Daduna • Erika Kaltenhäuser
allergy
IFN-γ Elispot
basophil activation
MISSION
in Kampala, Uganda. A total of 135 individuals were enrolled:
and remote infection in tuberculosis contact tracing. PLoS One.
We examine phenotypic and functional changes in 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 infectious disease patients as well in allergy patients.
109 with a new diagnosis of HIV-1 infection but no active TB,
2010 Jul 20;5(7):e11670.PMID: 20652022
19 with HIV-1 infection and active TB, and seven HIV-uninfected
healthy subjects. In control subjects immune responses were
internal and external collaborations
positive in 57.2 % by TST and in 100 % by at least one interferon-
Mycobacterial infections related projects are performed in close
gamma release assay. In HIV-1 infected patients without active
internal collaboration with the Division of Infectious Diseases
most important findings
and remote infection in tuberculosis contact tracing. (in
TB, induration in the TST (mm) (rho = 0.41, p-value <0.0001) and
(C.Lange) and the Outpatient Clinic (U.Greinert) and external
Antimycobacterial immune responses in patients with
collaboration with the Division of Infectious Diseases [C. Lange
concentration of interferon (IFN)-g in the QFT-G-IT tube with
collaborations with members of the TBNET.
pulmonary sarcoidosis.
et al.])
Mycobacterium tuberculosis-specific antigens (rho = 0.38; p =
Allergy related projects are performed in close collaboration
(in collaboration with the Division of Infectious Diseases [C.
IFN-g release assays (IGRA) have improved tuberculosis contact
0.0001) were negatively correlated to numbers of circulating
with the Division of Clinical and Molecular Allergology (U.Jappe,
A.Petersen)
Lange et al.])
tracing, but discrimination of recent from remote Mycobacterium
CD4+ T-cells, while numbers of IFN-g producing cells (rho = 0.03-
Sarcoidosis is a multisystem granulomatous disease of
tuberculosis contacts is not possible by IGRA alone. We present
0.13; p-value = 0.21-0.77) and frequencies of positive test results
The project C6 (Sleep supports the adaptive immune response to
unknown origin. Pathogenetic involvement of Mycobacterium
results of a tuberculosis contact investigation with a new ESAT-
for the T-Spot.TB test among groups of patients with different
vaccination) of the Collaborative Research Center 654 Plasticity
tuberculosis has frequently been discussed in the aetiology of
6 and CFP-10 specific interleukin (IL)-2 ELISpot in addition to
levels of immunodeficiency remained constant (p-value = 0.46).
and Sleep is jointly headed by T. Lange (UKSH Campus Lübeck)
and M.Ernst (Division of Immune Cell-Analytics, Borstel)
sarcoidosis; however, studies still remain contradictory.
ESAT-6 and CFP-10 specific IFN-g ELISpot and tuberculin skin
In HIV-1 infection, TST and QFT-G-IT immune responses are
We addressed the question of mycobacterial involvement in
testing (TST). Results of the TST, IFN-g ELISpot and IL-2 ELISpot
both strongly related to the degree of immunodeficiency, while
the pathogenesis of sarcoidosis by analysing cellular immune
were positive in 6/172 (3.4 %), 7/167 (4.2 %) and 6/196 (3.1 %)
results of the T-Spot.TB are independent of the level of CD4+
responses to mycobacterial antigens.
of contacts, respectively. Close contact (>or =100 hours) to the
T-cell depletion.
We examined the interferon (IFN)-g production by enzyme-
index case increased the risk of positive results in the IFN-g
linked immunospot in response to purified protein derivate
ELISpot, TST, and IL-2 ELISpot by 40.8, 19.3, and 2.5 times,
selected puBlications
(PPD) mycobacterial-specific antigen early secretory antigenic
respectively. Individuals with a positive IFN-g ELISpot/negative
Hörster R, Kirsten D, Gaede KI, Jafari C, Strassburg A,
target (ESAT)-6 and culture filtrate protein (CFP)-10 by peripheral
IL-2 ELISpot result had a median (IQR) duration of index case
Greinert U, Kalsdorf B, Ernst M and Lange C. Antimycobacterial
Collaborative Research Center 654 Plasticity and Sleep
Project C6 (together with T. Lange, UKSH Campus Lübeck) Sleep
blood mononuclear cells (PBMC) and bronchoalveolar-lavage
exposure of 568 hours (133_1000) compared to individuals
immune responses in patients with pulmonary sarcoidosis. The
mononuclear cells (BALMC) of patients with pulmonary
with a positive IFN-g ELISpot/positive IL-2 ELISpot result
Clinical Respiratory Journal 2009; 3: 229–238.
sarcoidosis, smear-negative tuberculosis and controls.
(median = 24 hours; 20_130; p-value = 0.047). Combination of
Release of IFN-g in response to ex vivo contact with PPD, ESAT-
a M. tuberculosis specific IFN-g ELISpot with a M. tuberculosis
Jafari C, Thijsen S, Sotgiu G, Goletti D, Benítez JA, Losi
6 or CFP-10 by BALMC and PBMC were comparable among
specific IL-2 ELISpot significantly improved the identification
M, Eberhardt R, Kirsten D, Kalsdorf B, Bossink A, Latorre
patients with sarcoidosis and controls (PBMC P = 0.23; BALMC
of individuals with the highest risk of recent M. tuberculosis
I, Migliori GB, Strassburg A, Winteroll S, Greinert U,
P = 0.18) and were less frequently observed in both groups
infection and is a promising method that should be explored to
Richeldi L, Ernst M, Lange C; Tuberculosis Network European
compared to patients with tuberculosis (BALMC P< 0.05; PBMC
target tuberculosis preventive chemotherapy.
Trialsgroup. Bronchoalveolar lavage enzyme-linked immunospot
patients differed from that of patients with tuberculosis, as well
Relationship of immunodiagnostic assays for tuberculosis
European Trialsgroup study. Am J Resp Crit Care 2009 Oct 1;
as from that of controls, while within BALMC it resembled that
and numbers of circulating CD4+ T-cells in HIV infection.
180(7):666-73.
of patients with tuberculosis.
(in collaboration with the Division of Infectious Diseases [C.
P< 0.0001). Within PBMC, the immunophenotype of sarcoidosis
Grant support
for a rapid diagnosis of tuberculosis: a Tuberculosis Network
Leidl L, Mayanja-Kizza H, Sotgiu G, Baseke J, Ernst M,
CONCLUSION: In contrast to patients with tuberculosis, the
Lange et al.]), Tuberculosis Research Unit, Case Western
frequency of mycobacteria-specific local and systemic immune
Reserve University, Cleveland, OH, USA [Z.Toossi] and Institute
Hirsch C, Goletti D, Toossi Z, Lange C. Relationship of
responses is not elevated in patients with sarcoidosis when
of Infectious Diseases, Makerere University, Kampala, Uganda
immunodiagnostic assays for tuberculosis and numbers of
compared to controls. The immunophenotype represents the
[H. Mayanja-Kizza and J. Baseke]
circulating CD4+ T-cells in HIV infection. European Respiratory
local resemblance of the granulomatous reaction underlying
Infection with HIV is the greatest risk factor for tuberculosis (TB)
Journal 2010 Mar;35(3):619-26.
tuberculosis and sarcoidosis while showing systemical
in Africa. Tuberculin skin test (TST), QuantiFERON-TB Gold In-
difference. These observations do not support a role of an
Tube (QFT-G-IT) and T-Spot.TB assays were performed in newly
Krummel B, Strassburg A, Ernst M, Reiling N, Eker B, Rath H,
infection with M.tuberculosis in the pathogenesis of sarcoidosis.
diagnosed HIV-infected individuals with and without active TB
Hoerster R, Wappler W, Glaewe A, Schoellhorn V, Sotgiu G,
Potential role for IL-2 ELISpot in differentiating recent
and in HIV-uninfected subjects at a university outpatient clinic
Lange C. Potential role for IL-2 ELISpot in differentiating recent
supports the adaptive immune response to vaccination
51
Dept. immunology and cell Biology • Immune Cell-Analytics
Dr. Martin Ernst
Innate Immunity
Innate immunity
Suhad Al-Badri • Ina Goroncy • Kimberley Kallsen • Jenni-Marie Ratten • Prof. Dr. Thomas Roeder (Guest
Professor) • Dr. Thomas Scholzen • Katrin Sprenger • Karina Stein • Karin Uliczka • Prof. Artur J. Ulmer
allergy
PD Dr. Holger Heine
MISSION
can be activated independently of the pathogen-responsive im-
with COPD, but upregulated upon S. aureus colonization. BMC
The mission of the division of Innate Immunity is the investigation, analysis and characterization of activation mechanisms of the innate immune system through microbes, microbial structures and allergens 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, asthma and COPD. Stress induced activation of the transcription factor FoxO and
its cross regulation with the innate immune reactions in the remost important findings
Allergy and asthma: LPS is the allergy-protective principle spiratory epithelium of the fruit fly Drosophila melanogaster (in
collaboration with Prof. Thomas Roeder, CAU Kiel)
of Acinetobacter lwoffii F78
munity pathways. (supported by DFG, SFBTR22, project A07).
Pulmonary Medicine 2011, 11:10.
Histone Deacetylase 1 modulates the expression of human beta
defensin 1 in lung epithelial cells
TLR4/lipid A
Human beta defensin 1 (DEFB1) is an important component of
antagonist
the innate immune response.
compound
This antimicrobial peptide is constitutively expressed and not
406 blocks
directly regulated by bacterial triggers or cytokines. The deregu-
the Acineto-
lation of its expression has been implicated in the pathogen-
bacter lwoffii
esis of several diseases like COPD or asthma. However, the
F78-induced IL-
been identified In numerous epidemiological studies to strongly
The innate immune system represents an ancient host defense
molecular mechanisms that regulate DEFB1 gene expression
12p70 release
influence the development of allergic reactions later in life.
mechanism that protects against invading pathogens. An impor-
remain unknown. In this study we investigated whether epigen-
in human
Moreover, it had been shown that certain bacteria isolated
tant class of immune effector molecules to fight microbial infec-
etic processes are involved in the regulation of the basal con-
moDCs.
from this environment have allergy-protective properties in
tions are antimicrobial peptides (AMPs) which are conserved
stitutive expression of DEFB1 in lung epithelial cells (A549). Our
mouse allergy models (Debarry et al, JACI 2007). In the present
over a wide variety of species. In the fruit fly Drosophila mela-
data demonstrate a role of histone deacetylases (HDACs) in
Figure 2:
study, we further characterized one of these bacteria, namely
nogaster just like in other organisms, the induction of AMPs in
the regulation of DEFB1 gene expression. In humans there are
Increased
the Gram-negative Acinetobacter lwoffii F78, with regard to the
response to infection is regulated by NF-κB transcription factors
11 HDACs subdivided into four classes. Using isoform-selective
activation of
bacteria-induced signaling and possible mechanisms of allergy
and the activation of the immune deficiency (IMD) and Toll path-
inhibition, knock-down by siRNA and overexpression of HDACs
antimicrobial
protection. The impact of A. lwoffii F78 on human monocyte-de-
ways, homologues of the human tumor necrosis factor pathway
we show that HDAC1 represses DEFB1 gene expression in lung
peptides as a
rived dendritic cells especially with respect to their T helper cell
(TNFR) and the Toll-like receptor 4 pathway, respectively.
epithelial cells. ChIP assays additionally reveal that inhibition of
consequence of
polarization capacity was investigated by ELISA, FACS analysis
Recent studies show that AMP activation in Drosophila can be
HDACs results in modifications of the chromatin at the DEFB1
oxidative stress
and real-time PCR experiments as well as confocal microscopy.
achieved independently of these immunoregulatory pathways
promoter. Histone modifications associated with transcriptional
in a AMP/
We could show that A. lwoffii F78 induces a TH1-polarizing pro-
by a member of the forkhead box class O transcription factor
activation, H3 acetylation and H3K4 trimethylation, were in-
GFP-expressing
gram in human dendritic cells, including the upregulation of TH1-
(FoxO), a key regulator of stress resistance, metabolism and
creased after inhibition of HDAC1-3. Thus our data suggest that
Drosophila.
driving molecules such as ICAM-1, IL-23p19 or CCL5 as well as
ageing. FoxO is an important mediator of the insulin signaling
HDAC1 transcriptionally represses DEFB1 by reducing chromatin
accessibility at the DEFB1 promoter.
the induction of IFN-γ release by co-cultured autologous naÏve T
pathway, adjusting growth and metabolism to nutrient availabil-
cells. In addition, TIM-3 expression was induced and the TBet/
ity. A FoxO-dependent AMP up regulation can be induced by
GATA3 transcription factor expression ratio was shifted towards
hunger via the insulin/insulin-like growth factor signaling path-
selected puBlications
internal and external collaborations
TH1. Blocking experiments using anti-TLR2 antibodies and lipo-
way (ILS) especially in barrier tissues.
Debarry, J., A. Hanuszkiewicz, K. Stein, O. Holst, and H.
Otto Holst, Ulrich Zähringer, Helmut Brade (Molecular Infectiol-
polysaccharide (LPS)/TLR4-antagonists (Fig.1) indicated that the
Since we have mainly focused on the investigation of the innate
Heine. 2010. The allergy-protective properties of Acinetobacter
ogy, Research Center Borstel)
LPS of A. lwoffii F78 was the responsible molecule promoting
immune reactions in the fly’s epithelial tissue of the respiratory
lwoffii F78 are imparted by its lipopolysaccharide. Allergy
Thomas Roeder, CAU Kiel
these effects. Moreover, isolated LPS from A. lwoffii could fully
organs, we concentrated on a closer characterization of the
65:690-697.
Peter König, University Lübeck
substitute for whole bacteria regarding all investigated effects
FoxO-dependent AMP activation in these organs. We could
Harald Renz, Holger Garn, University Marburg
of the activation. In addition, we are currently investigating epi-
demonstrate a translocation of FoxO into the epithelial cell nu-
Brade, L., H. Heine, S. Raina, G. Klein, F. di Padova, H.
genetic control mechanisms for the driving of TH1 polarization,
cleus not only as a result of nutritional deficiency but also as a
Brade, and S. Müller-Loennies. 2011. Immunization with an an-
in particular the effect of different histone deacetylases.
In summary, we found evidence that the allergy-protecting ef-
Erika von Mutius, LMU Munich
response to environmental stress factors, such as heat, cold, UV
ti-idiotypic antibody against the broadly LPS-reactive antibody
Grant support
radiation and oxidative stress, which also leads to the upregula-
WN1 222-5 induces E. coli R3-core-type specific antibodies in
SFB/TR22, project A02
tion of AMPs (Fig.2).
rabbits. Innate Immunity (accepted)
fects of A. lwoffii F78 are due to the activation of a TH1-polar-
DAAD PPP Ägypten: “Immunomodulatory effects of probiotic
bacteria”
izing program in human dendritic cells, and that the LPS of A.
Using the advantages of Drosophila as a model organism and
Andresen, A., C. Lange, D. Strodthoff, T. Goldmann, N.
lwoffii F78 is responsible for these beneficial effects (supported
investigative tool we detected new mechanisms of cross-regu-
Fischer, H. Sahly, D. Branscheid, and H. Heine. 2011. S100A7/
by DFG, SFB/TR22, project A2).
lation of metabolism and innate immunity by which AMP genes
psoriasin expression in the human lung: unchanged in patients
53
Figure 1:
Dept. immunology and cell Biology • Innate Immunity
Exposure to farming environment during early childhood has
COPD
Fluorescence Cytometry
flow cytometry
training
cell sorting
live cell imaging
microscopy
MISSION
analysis of up to nine fluorescence channels plus two scatter
Nottingham, UK) on nuclear trafficking of IPSE/alpha-1 was
The research strategy of the FZB relies on the comprehension of molecular and cellular interactions in the fields of inflammation, infection and allergy. The analysis of such processes requires the ability to phenotypically characterize individual cells concerning the expression of surface, intracellular and secreted molecules. The unit Fluorescence Cytometry was established in spring 2008 and offers scientists at the FZB and at associated universities access to modern fluorescence-based techniques by providing both, state-of-the-art flow cytometers as well as confocal and wide-field fluorescence microscopes. In addition, the facility carries out service and offers training for the instruments. The service unit is also available for consultation about the design and data analysis of individual experiments. Demanding experimental strategies can lead to joint research projects.
parameters is possible.
initiated together with the division of Cellular Allergology.
Scientifically, the unit Fluorescence Cytometry is associated with the division of Innate Immunity (Head PD. Dr. Holger Heine). The Fluorescence Cytometry unit contributes to the projects of this division by applying advanced live cell imaging and flow cytometry techniques to scientific problems in the fields of innate immunity and allergy.
course comprises three full days of training including theory
Training
Figure 1:
Technical training is an integral part of the Fluorescence
The FACSAria
Cytometry concept. Training for the BD LSR II and the Olympus
cell sorter
IX-81 is provided on an individual basis depending on the
particular projects. The basic training for the Leica SP5 consists
of a 2-3 hour introduction to the instrument. Since April 2008
approx. 56 scientist and technicians were instructed in this
way. The training for cell sorting is more demanding: The basic
and practical instructions in small groups. Since October 2008
16 colleagues were qualified to run the BD FACSAria.
Research
TECHNOLOGY
Leica TCS SP5 offers advanced optical capabilities. Samples
Research projects of the Fluorescence Cytometry unit are
The mission of the FZB is focused on lung diseases caused either
can be excited by nine different laser lines ranging from 405
accomplished in collaborations with laboratories in and outside
by infectious agents or evoked by aberrant immunomodulations
to 633 nm which can be adjusted independently by acousto-
the FZB (see below). In close association with the division of
leading to inflammation and allergy. To understand these
optical tunable filters (AOTF). Due to spectral detection by
Innate Immunity the interplay of innate and adaptive immune
processes in detail, it is necessary to study the recognition
five fluorescence detectors and the use of an acousto-optical
responses in the context of infectious and allergic diseases
and uptake of pathogens/allergens, the modulation of host
beam splitter (AOBS). emission filters and dichroic mirrors are
is studied. An ongoing project is aimed at defining the role
cell function, the intercellular communication and the induced
not needed. Instead, the optical properties can be instantly
of autophagy and autophagy constituents in the immune
signal transduction cascades in cell and tissue culture systems
set via the control software. This feature makes the system
response to Lactococcus lactis. Previous experiments suggest
as well as in vivo animal models. Using fluorescence-based
extremely flexible with regard to the applicable dyes. For high
that autophagy may play a crucial role in the IL-12 response of
techniques it is possible to characterize individual cells with
frame rates an additional resonant scanner is present. The
dendritic cells to this bacterium. Using (live-) cell imaging and
regard to the expression of surface and intracellular molecules
microscope contains an incubation chamber with temperature
biochemical approaches, the role of individual components
(e.g. receptors, transcription factors, second messengers),
regulation and CO2 gassing. The system can be used for
of the autophagy machinery for the processing and signal
secreted components (e.g. cytokines and defensines) or the
conventional image analysis as well as live cell imaging
transduction of L. lactis is elucidated (Fig. 2).
direct interaction with other cells. The Fluorescence Cytometry
including FRET and FRAP experiments.
internal and external collaborations
unit employs two, in part complementary, key technologies to
Flow Cytometry
The Fluorescence Cytometry unit is scientifically associated with
Flow cytometry allows to analyze large numbers of cells in a
the division of Innate Immunity. Further in-house collaborations
quantitative manner. Moreover, employing FACS sorters, it is
comprise the investigation of Fas (CD95) death receptor
High resolution imaging is a key technique to obtain
possible to highly purify individual populations of living cells
trafficking and sorting in co-operation with the Division of
information about the spatial distribution of components and
for subsequent experiments. The BD LSR II flow cytometer
Molecular Immunology (Head Dr. Kyeong-Hee Lee) and a
the chronological sequence of events. For live cell analysis,
is especially well equipped for multiparameter analysis.
joined project with Dr. Gabriele Schramm (Division of Cellular
the Olympus IX-81 imaging system is available. It features an
With its fixed-alignment optics and digital electronics the
Allergology; Head PD. Dr. Helmut Haas) on the trafficking of
incubation chamber with CO2 gassing as well as temperature
simultaneous analysis of up to eight fluorescence channels
IPSE/alpha-1 in primary cells. An external collaboration is
and humidity regulation. With four fluorescence channels, a
plus two scatter parameters is possible. The BD FACSAria flow
carried out with Dr. Elena Csernok (Clinical Center Bramstedt)
accomplish such analyses.
Fluorescence Microscopy
highly sensitive CCD camera and a motorized stage it is well-
cytometer is a high speed “benchtop” cell sorter (Fig. 1). Up
on neutrophil cytoplasmic auto-antibodies (ANCA) and an
suited for time lapse imaging. For demanding experimental
to 70,000 events per second can be acquired. With its fixed-
international co-operation with Prof. Dr. Franco H. Falcone (The
strategies, the inverse confocal laser scanning microscope
alignment optics and digital electronics the simultaneous
School of Pharmacy, Division of Molecular and Cellular Science,
Figure 2:
Uptake of
Lactococcus
lactis by human
dendritic cells
(DNA, red; lysosomes, blue;
autophagy
marker protein
LC3, green)
55
Dept. immunology and cell Biology • Fluorescence Cytometry
Dr. Thomas Scholzen
Inflammation and Regeneration
Prof. Dr. Heinz Fehrenbach
nanoparticles
emphysema
animal models
inflammation
Franziska Ganzert • Dörte Grella • Jennifer Klein • Lars Lunding • Dr. Sandra Schlick • Dr. Christina
Vock • Dr. Michael Wegmann
MISSION
is the clinical observation that recurrent asthma exacerbations
External collaboration has been established with the groups
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 COPD/emphysema, allergic bronchial asthma, and inflammatory processes related to occupational exposures.
triggered by viral (Coxsackie and Parainfluenza virus) or bacte-
of Henning Bockhorn (Karlsruhe Institute of Technology), Bernd
rial (Haemophilus influenza) infections are major risk factors for
Müller (Philipps-University of Marburg), Tanja Hansen (Fraun-
asthma aggravation towards a severe phenotype. Therefore, in
hofer Institute of Toxicology and Experimental Medicine, Han-
an already established mouse model of chronic asthma repeat-
nover) and Peter König (University of Lübeck) to investigate cell
most important findings
patients, short-term stimulation of human tissue samples with
ed exacerbations of the asthmatic phenotype are evoked by
type- and compartment-specific toxicological effects of lung ex-
Pulmonary emphysema is characterized by the destruction of
palifermin are used in co-operation with the division of clinical
intra-nasal application of poly-I:C, a ligand of Toll-like receptor 3
posure with tailor-made Carbon Black nanoparticles.
alveoli with ensuing airspace enlargement and loss of alveolar
and experimental pathology. Preliminary data suggest that pali-
(TLR3) which responds to RNA virus infection. Whereas poly-I:C
gas-exchange area, which is associated with severe impairment
fermin is indeed able to stimulate human lung epithelial cells
alone induced neutrophilic inflammation in the bronchoalveo-
of oxygen uptake in emphysema patients. Curative therapeutic
(Figure 1). Unlike mouse lung tissues, however, stimulation may
lar lavage of “healthy” mice, a mixed eosinophilic/neutrophilic
options are still lacking, which may explain why emphysema is
not be exclusive of lung epithelium.
inflammation was induced by poly-I:C in OVA-asthmatic mice.
Because co-activation of dendritic cells by TLR3 ligands together
still the most frequent indication for single lung transplantation.
The induction of regeneration of lost alveolar tissues is still a
Severe bronchial asthma developed into a major health prob-
with epithelium-derived thymic stromal lymphopoietin (TSLP) can
major challenge in the development of novel curative therapies
lem because many patients suffering from this disease exhibit
trigger IL-23 production, which is required for Th17 cell differentia-
for emphysema.
a high degree of resistance towards corticosteroid treatment.
tion, epithelial expression of TSLP is currently under investigation.
There is increasing evidence that patients characterized by a
Therefore, we have tested if the local therapeutic treatment of
neutrophilic inflammatory phenotype are particularly prone to
selected publications
the emphysematous mouse lung with N-terminally truncated re-
the development of severe asthma and display corticosteroid
Yildirim A, Muyal V, John G, Muller B, Seifart C, Kasper M,
combinant human keratinocyte growth factor (∆N23-KGF, palifer-
resistance more frequently. The recruitment of neutrophils (es-
Fehrenbach H. Palifermin induces alveolar maintenance pro-
min), which is a strong mitogen of lung epithelial cells, is able
pecially in the absence of bacterial or viral infection) remains
grams in emphysematous mice. AMERICAN JOURNAL OF RESPI-
to induce alveolar regeneration (Yildirim et al. AJRCCM 2010).
incompletely understood. Several cell types including bronchial
RATORY AND CRITICAL CARE MEDICINE 2010;181:705-17
We used elastase-instillation into mouse lungs to induce severe
epithelial cells are capable of producing neutrophilotactic me-
lung emphysema within a few weeks to model the disease. To
diators. In addition, a new type of T helper (Th) 17 cells may
Vock C, Hauber H, Wegmann M. The other T helper cells in
test for the regenerative effect of palifermin, such emphysema-
link T cell-driven airway inflammation and neutrophilc influx
asthma pathogenesis. JOURNAL OF ALLERGY 2010;2010:519298
tous lungs were therapeutically treated, i. e. after establishment
in severe asthma. Hypothesis-driven studies in severe asthma
of the disease, via the airways at three occasions with 10 mg/
research are greatly limited by the fact that a suitable in-vivo
epithelium (red arrowheads) express transcription factor Pak4 protein.
Abram M, Wegmann M, Fokuhl V, Sonar S, Luger E, Kerzel S,
kg body weight. The main findings were that in mice treated
mouse model mimicking the specific disease characteristics of
Radbruch A, Renz H, Zemlin M. Nerve growth factor and neu-
with palifermin versus PBS i) airflow limitation associated with
this phenotype are still lacking.
rotrophin-3 mediate survival of pulmonary plasma cells during
emphysema was largely reversed as assessed by non-invasive
head-out body plethysmography and ii) elastase-induced air-
Figure 1: HOPE embedded human lung tissue incubated ex-vivo for
1 day with ∆N23-KGF (10ng/ml). Macrophages (blue arrowhead),
the Allergic Airway Inflammation. JOURNAL OF IMMUNOLOGY
Aiming at the development of such a model of severe asthma,
2009;182:4705-12
space enlargement as well as the loss of alveoli was partially
we succeeded to establish a model of antigen-induced airway
reversed as assessed by design-based stereology. The investi-
neutrophilia in the mouse. By using intra-peritoneal injection
gation of lung tissues from palifermin-treated emphysematous
of a model allergen (ovalbumin; OVA) adsorbed to complete
Collaboration within the Research Center Borstel has been es-
mouse lungs by means of RT-PCR and Western blotting supports
Freund’s adjuvant it was possible to establish a strong Th1 cell-
tablished with the Division of Clinical and Experimental Pathol-
the notion that palifermin induced important alveolar mainte-
directed immune response against OVA. Subsequent short term
ogy to investigate e.g. the effects of palifermin on human lung
Figure 2: Mouse model of antigen-induced airway neutrophilia.
internal and external collaborations
nance programs (e. g. VEGF-, TGFβ-pathways), which address
exposure to aerosolized OVA together with IL-18 (to reactivate
tissue in the short-term stimulation in-vitro model together with
Airway wall infiltration with neutrophilic granulocytes (arrow) revealed
the major structural components of the alveolar wall and pro-
Th1 memory cells) resulted in pronounced infiltration of neutro-
the group of Torsten Goldmann.
by immunohistochemistry for Ly6G.
mote alveolar regeneration. Further in-vitro studies in cell cul-
phils into the airways (Figure 2) associated with mucus hyper-
Several research projects are pursued in cooperation with
Grant support
tures systems suggest that the beneficial effects of palifermin
secretion and the development of airway hyperresponsiveness
groups at the University of Lübeck: studies of the airway gen-
on the interstitial tissue compartment are linked to active TGFβ-1
resembling three major hallmarks of human severe asthma.
eration-specific epithelial expression of IL-13 receptors together
DFG: Cluster of Excellence “Inflammation at Interfaces” (EXC306-
released by alveolar epithelial type II cells, the major target
This new mouse model is currently used as a basis for gener-
with Kathrin Kalies and Peter König, Institute of Anatomy; studies
DW2, EXC306-STI)
of ∆N23-KGF in lung parenchyma. In order to evaluate if these
ating a model of chronic antigen-induced airway neutrophilia.
on the effects of the proteasome inhibitor Bortezomib together
BMBF: Consortium “Carbon Black” (FKZ 03X0093A), coordina-
results observed in mice can be translated into emphysema
The starting point of a second approach we currently pursue
with Rudolf Manz, Institute for Systemic Inflammation Research.
tor: H.F., RCB
57
Dept. Pneumology • Inflammation and Regeneration
allergic bronchial asthma
Cellular Pneumology
Lung innate immunity
Beate Höschler • Linda Lang • Alexandra Lange • Dr. Christina Moulakakis • Vicky Sender
Rab GTPases
inflammation
soluble C-type lectins
Prof. Dr. Cordula Stamme
endolysosomal trafficking
MISSION
Our objective is to understand lung-specific innate imprinting by human surfactant protein (SP)-A and SP-D to Gram-negative bacteria and M. tuberculosis in order to develop soluble C-type lectin (sCTL)-based antimicrobials and immunotherapeutics. The specific activation and conversion mechanisms of small GTPases by sCTL to develop strategies that selectively target up- or downmodulation of endocytic transport, the role of sCTL in signal propagation derived from endocytic organelles, or the bacterial subversion of sCTL-determined NF-κB/IκB-α activity in living macrophages will provide a platform to approach this aim.
function of Rab7b. In AM and RAW264.7 macrophages, the
The pulmonary collectin surfactant protein (SP)-A has been
SP-A-enhanced lysosomal delivery of GFP-E.coli is abolished
implicated in modulating immune host defense responses in
by the inhibition of Rab7 and Rab7 small interfering (si)RNA
the lung in vivo and in vitro. SP-A directly interacts with alveolar
transfection, respectively. The constitutive expression of Rab7
macrophages (AM), the most abundant immunocompetent cells
in AM from SP-A -/- mice is significantly reduced compared with
in the alveolar space that form the first line of cellular defense.
SP-A+/+ mice and is restored by SP-A (Fig. 2). Rab7 blocking
Several studies demonstrated that SP-A uptake by AM occurs via
peptides antagonize SP-A-rescued lysosomal delivery of GFP-E.
selected puBlications
clathrin-mediated endocytosis (CME), and that SP-A-containing
coli in AM from SP-A -/- mice. Activation of Rab7, but not Rab7b, by
Moulakakis C, Stamme C. Role of clathrin-mediated endo-
vesicles traffick through the endolysosomal pathway. We have
SP-A depends on the PI3K/Akt/protein kinase C (PKC) z signal
cytosis of surfactant protein A by alveolar macrophages in
shown previously that CME of SP-A by AM is a prerequisite for
transduction pathway in AM and RAW264.7 macrophages.
intracellular signaling. AMERICAN JOURNAL OF PHYSIOLOGY
SP-A-mediated IkB-a stabilization, inhibition of LPS-induced NF-
SP-A induces a Rab7/PKCz interaction in these cells and the
– LUNG CELLULAR AND MOLECULAR PHYSIOLOGY. 2009 Mar;
kB activation and atypical (a)PKCz activation, and thus for its
disruption of PKCz by siRNA knockdown abolishes the effect
296(3):L430-41.
anti-inflammatory immunomodulation. The aim of the present
of SP-A on Rab7. The data demonstrate a novel role for SP-A in
Figure 2: SP-A-enhanced lysosomal delivery of GFP-E.coli in AM from
SP-A -/- mice involves Rab7. ∗∗p < 0.05; ∗∗∗p < 0.001 (SP-A-treated vs
untreated cells); ###p < 0.001 (C57Bl/6J vs SP-A -/- mice).
study was to determine the mechanism(s) of SP-A-mediated
modulating endolysosomal trafficking via Rab7 in primary AM
Sender V, Moulakakis C, Stamme C. Pulmonary Surfactant
endosomal signaling, and to identify scaffold proteins and
and define biochemical pathways involved.
Protein A Enhances Endolysosomal Trafficking in Alveolar
kinases involved in SP-A-specific modulation of AM signaling.
Macrophages through Regulation of Rab7. JOURNAL OF
By using RNA interference, we here show that knockdown
IMMUNOLOGY. 2010 Dec in press.
of clathrin heavy chain inhibits SP-A uptake (Fig. 1), and
significantly reduces SP-A-mediated IkB-a protein expression. In
internal and external collaborations
addition, clathrin knockdown abolishes SP-A-mediated inhibition
Inhouse collaborators include Dr. N. Reiling (Microbial Interface
of LPS-induced TNFa release, and abrogates aPKCz activation
Biology), C. Steinhäuser (Microbial Interface Biology), and Dr. C.
implicating CME as a major mechanism contributing to SP-A
Alexander (Immunochemistry).
anti-inflammatory imunomodulation.
National collaborators are Prof. Dr. C. Hübner (Dept. of Physics,
Univ. Lübeck), Prof. Dr. B. Müller (Div. of Pneumology, Univ.
Enhanced phagocytosis and endocytosis are key functional
Marburg), Prof. Dr. U. Costabel/Dr. F. Bonella (Ruhrlandklinik,
consequences of AM/SP-A interaction suggesting a SP-A-
Univ. Essen).
mediated modulation of small Rab GTPases that are pivotal
International cooperations were contracted with Dr. A. Kirby
membrane organizers in both processes. Here, we show that
(Univ. of York, UK) and Prof. Dr. S. Hawgood (Univ. of California,
SP-A specifically and transiently enhances the protein expression
San Francisco, USA).
of endogenous Rab7 and Rab7b, but not Rab5 and Rab11, in
primary AM from rats and mice. SP-A-enhanced GTPases are
Grant support
functionally active as determined by increased interaction of
Work in this group was supported by the Deutsche
Rab7 with its downstream effector Rab7 interacting lysosomal
Forschungsgemeinschaft (Grant 609/1-4 and 609/2-1 to C.S.;
protein (RILP) and enhanced maturation of cathepsin-D, a
Figure 1: Clathrin HC (CLTC) knockdown abrogates SP-A uptake.
Grant 1999/2-1 to C.M.).
Dept. Pneumology • Cellular Pneumology
most important findings
59
Pathophysiology of Inflammation
Acute lung injury
chronic inflammatory lung disease
granuloma
Dr. Markus Blaukovitsch • PD Dr. Daniel Droemann • PD Dr. Karoline I. Gaede • Eike Hansen • Jessica
Hofmeister • Romina Pritzkow • Marius Schawaller • Carsten Sparke • Lisa Sprenger • Christoph Zabel
Prof . Dr. Peter Zabel
parenchymal lung disorder
mucus hypersecretion
ventilation
MISSION
Vollmer E, Zabel P. BMC Pulm Med. 2010 Apr 21;10:21.
The reserach group „Pathophysiology of Inflammation“ is interested in pathomechanisms of acute and chronic pulmonary inflammation and the resultant functional impairment. Work mainly focuses on COPD, bronchial asthma, pneumonia, and chronic granulomatous diseases (eg. sarcoidosis, chronic beryllium disease). Moreover, ventilator-induced injury (VILI) and ARDS are main topics of the group. Our goal is to identify possible treatment targets that may be transferred to clinical use.
Figure 1:
Inhibition of the
Comparison of the effect of LPS and PAM3 on ventilated lungs
calcium-acti-
Hauber HP, Karp D, Goldmann T, Vollmer E, Zabel P. BMC
vated chloride
Pulm Med. 2010 Apr 20;10:20
channel hCLCA1
attenuates
Association of IBD Risk Loci with Sarcoidosis and its Acute and
bacterial-
most important findings
In another set of experiments we intratracheally applied
Chronic Subphenotypes. Fischer A, Nothnagel M, Franke A,
induced mucus
Mucus hypersecretion is an important clinical symptom in
bacterial production to investigate the effect of ventilation and
Jacobs G, Saadati HR, Gaede KI, Rosenstiel P, Schürmann
expression
chronic inflammatory lung diseases. During the past years our
another injury (two hit model) on lungs. We found that products
M, Müller-Quernheim J, Schreiber S, Hofmann S. Eur Respir J.
in explanted
group established an ex vivo model of explanted human airway
from Gram-negative bacteria can further enhance ventilator-
2010 Jul 22. [Epub ahead of print]
mucosal
mucosa to study the pathophysiology of mucus hypersecretion.
induced inflammation. In contrast, products from Gram-positive
This model enables us to investigate both epithelium and
bacteria did not only increase ventilator-induced inflammation
The TGF-beta-pseudoreceptor BAMBI is strongly expressed
medium alone.
underlying mucosa with inflammatory cells. We were able to
but also induced changes in lung function parameters. This
in COPD lungs and regulated by nontypeable Haemophilus
B: stimulation with LPS (lipopolysaccharide). C: stimulation with LPS
describe the effect of different bacteria (eg. Pseudomonas
finding may have important clinical implications since ventilator-
influenzae. Drömann D, Rupp J, Rohmann K, Osbahr S,
and the calcium channel inhibitor niflumic acid. D: stimulation with LPS
aeruginosa), bacterial products (eg. LPS, PAM3) and cytokines
associated pneumonia is most often caused by Gram-positive
Ulmer AJ, Marwitz S, Röschmann K, Abdullah M, Schultz H,
and MSI-2216 (an analogue of niflumic acid). PAS staining. Original
(eg. TNFa, Th2 type cytokines) on mucin expression. Using
Staphylococcus aureus. Recently, we used the mouse model of
Vollmer E, Zabel P, Dalhoff K, Goldmann T. Respir Res. 2010
magnification x100.
several
glucocorticosteroids,
bleomycin-induced lung fibrosis to study the effect of mechanical
May 31;11:67
N-acetylcysteine, niflumic acid) we could inhibit mucin expression
ventilation in fibrotic lungs as another example of a two hit
pharmacologic
agents
(eg.
airway tissue. A:
Figure 2:
and analyze the effect of various drugs. Our experiments
model. By application of multiplex cytokine analysis we were
internal and external collaborations
Numbers of
showed that drugs like glucocorticosteroids that are often used
able to identify potential key cytokines in the pathogenesis of
Our group cooperates with the followingl research groups at
polymorpho-
in clinical practice can indeed attenuate bacterial- and Th2
VILI in fibrotic lungs. This is under current investigation.
the Research Center Borstel: Inflammation and regeneration,
nuclear cells
clinical and experimental pathology, immunochemistry, medical
(PMN) per high
cytokine-induced mucus expression. In another study we were
able to demonstrate inhibition of bacterial- but not Th2 type
Sarcoidosis and chronic beryllium disease are clinically
and biochemical microbiology, mucosa immunology, immune
powered field in
cytokine-induced mucus expression by N-acetylcyteine. With
indistinguishable with a heterogeneous spectrum of often
cell analytics. We work together in projects with the UKSH
the mouse lung
our findings we provide a rational molecular basis for the use
unpredictable phenotypes. Both disorders are multifactorial,
Campus Lübeck and Campus Kiel, the university hospital of
of spontaneous-
of agents to treat mucus hypersecretion in clinical practice.
meaning that development of disease depends on genetic
Freiburg, the university of Giessen Lung Center (UGLC), the
ly breathing ani-
However, at present all pharmacologic treatment options are
and exogenous factors. Our research interests are genetic
university hospital Hamburg, German Network fro Diffuse
mals (w/o, without mechanical ventilation) and mechanical ventilated
mainly anti-inflammatory and not mucus specific. Further studies
susceptibility and understanding of pathogenesis and hereby
Parenchymal Lung Disease (GOLD-net), Genotype Phenotype
animals without challenge (control), with intratracheal saline challenge
are ongoing to identify therapeutic targets to selectively inhibit
improvement of diagnosis. Our aim is the identification of
Relationship in Sarcoidosis (GenPhenSarc), Medical Imaging
(sham) and with challenge with LPS or PAM3 at different doses. Bars
mucus hypersecretion.
differential and prognostic biomarkers to be used by clinicians
Electronics (MiE) GmbH Seth. International collaborations exist
indicate mean+SEM. *: P < 0.05 vs sham
including in practice applicable detection of Beryllium
with the Meakins-Christie Laboratories (Monteral) the university
VILI is an important clinical problem in intensive care medicine.
sensitisation.
To study the pathophysiologic mechanisms of VILI we
“Tor Vergata” (Rome), Sourasky Medical Center (Tel Aviv),
CDC, NIOSH (Morgantown), and the university of Pennsylvania
established a mouse model of invasive mechanical ventilation.
selected puBlications
We first evaluated the effect of different ventilation strategies
Dexamethasone and N-acetyl-cysteine attenuate Pseudomonas
on lung function and inflammation. Previous studies found
aeruginosa-induced mucus expression in human airways.
Grant support
that mechanical ventilation with high tidal volumes leads to
Sprenger L, Goldmann T, Vollmer E, Steffen A, Wollenberg
BMBF
inflammation and barrier dysfunction. We also looked at very
B, Zabel P, Hauber HP. Pulm Pharmacol Ther. 2010 Dec 10.
DFG
low volume ventilation and showed that very low tidal volume
[Epub ahead of print]
Industry (Bayer, ResMes)
ventilation also induces enhanced inflammation and worsening
of lung function parameters. This may be due to cyclic opening
Effect of low tidal volume ventilation on lung function and
and closure of alveoli.
inflammation in mice. Hauber HP, Karp D, Goldmann T,
(Phildelphia).
61
Dept. Pneumology • Pathophysiology of Inflammation
PD Dr. Hans-Peter Hauber
Barrier-Integrity
fluid balance
Dr. Heike Dombrowsky • Kathrin Handge • Jürgen Sarau • Tanja Steiner • Yuk Lung Wong
intestine
physiology
Ingmar Lautenschläger
MISSION
Medicine”, Department of Anesthesiology and Intensive Care,
In septic and other intensive care patients, mesenteric microcirculation and intestinal barrier function are critical for survival yet very difficult to maintain. Inflammation induces vasomotor and barrier dysfunction, ultimately leading to gut failure, loss of fluid and protein from the circulation and translocation of bacteria (remnants) to peritoneum and circulation. Our dedication is the exploration of molecular mechanisms involved and the identification of relevant therapeutic targets.
University Medical Center Schleswig-Holstein, Campus Kiel)
and Prof. S. Uhlig (Institute of Pharmacology and Toxicology,
In cooperation with PD. Dr. B. Lindner (Division of Immunochemistry, Research Center Borstel) we have examined the
the range of applications (e.g. availability of antibodies for im-
influence of dietary n-3 FA on the intestinal phospholipid com-
We have finished the technology development of a novel com-
munohistochemistry or use of k.o. strains for specific questions).
position by HPLC-MS. The state of the art technique identified
plex model system - the isolated perfused rat small bowel. It
A light microscope for preparation has been introduced, the
more than 40 different molecular species, demonstrating indivi-
enables us for the first time to analyze the intestinal patho-
preparation technique and perfusion rates have been modified
duality of phospholipid classes’ composition and high flexibility
physiology, in particular the intestinal barriers, in detail in a
and the perfusion chamber including cannulae and balances
of the intestinal lipid pool.
compartment related fashion. Using this model we have shown
has been adapted.
PD Dr. A. Frey (Division of Mucosal Immunology, Research Cen-
tor of inflammation induces vasomotor and barrier dysfunction
ter Borstel) cooperates with us to explore the processing of
as well as changes in fluid homeostasis likely seen in acute
antigens at mucosal surfaces. In these experiments, the isolated intestine provides an environment for the reproducible
intestinal failure.
application of antigens and identification of routes across the
intestinal epithelium.
Our dose-response studies of PAF and the pre-treatment with
antagonists of the eicosanoid, the sphingolipid as well as intracellular kinase pathways lead to the hypothesis that there
The goal of the collaboration with Dr C.E. Boyle (Department of
are different cellular signaling pathways responsible for both
Medical Microbiology, Virology and Hygiene, UKE, Hamburg) is
the microcirculatory disturbances and the barrier dysfunction
the exploration of the early interaction of yersinia - which cause
and that these pathways can be modulated by pharmacologic
enteritis or mesenteric lymphadenitis - with the intestinal mucosa. With a laboratory fulfilling the S2/L2 criteria we are able to
agents.
safely conduct isolated perfusion experiments with pathogenic
We completed a study on the effects of dietary n-3 fatty acids
Figure 1: Preparation of the isolated intestine
strains and genetically modified organisms. These experiments
will elucidate routes of yersinia invasion and thus provide in-
(FA) of PAF induced vasoconstriction and hyperpermeability.
Despite incorporation of n-3 FA into intestinal phospholipids
selected puBlications
and PAF induced eicosanoid generation we found no alleviati-
Lautenschläger I, Dombrowsky H, Frerichs I, Kuchenbecker
on of vasoconstriction and even elevated intestinal permeabi-
S, Bade S, Schultz H, Zabel P, Scholz J, Weiler N, Uhlig S. A
Grant support
lity in the isolated intestine. These data indicate that the bene-
model of the isolated perfused rat small intestine. AMERICAN
Medical Faculty of the Christian-Albrechts-University, Kiel
ficial anti-inflammatory effects of n-3 FA in critically ill patients
JOURNAL OF PHYSIOLOGY-GASTROINTESTINAL AND LIVER
Leibniz-Vorhaben im wettbewerblichen Verfahren im Rahmen
need to be balanced against the possibility of enhanced vas-
PHYSIOLOGY 2010 Feb;298(2):G304-G313
des Paktes für Forschung und Innovation
cular permeability as a side effect of fish oil supplementation.
Furthermore, they suggest - in line with the above mentioned
Dombrowsky H , Lautenschläger I, Zehethofer N, Lindner B,
findings - that mediators other than eicosanoids are responsib-
Schultz H, Uhlig S, Frerichs I, Weiler N. Ingestion of n-3 fatty
le for the immediate vasoconstriction and edema formation in
acids augments basal and platelet activating factor-induced rat
response to PAF and open up the possibility of novel therapeu-
intestinal permeability. JOURNAL OF NUTRITION (under review)
tic approaches.
internal and external collaborations
Another challenging project is the adaptation of the isolated
We collaborate extensively and discuss all projects with Prof.
perfused intestine model to the mouse anatomy, broadening
N. Weiler and Prof. I. Frerichs (working group “Intensive Care
sight in the pathophysiology of the disease.
Dept. Pneumology • Barrier-Integrity
Medical Faculty of RWTH Aachen University).
most important findings
that platelet activating factor (PAF) as an archetypical media-
63
National Reference Center for Mycobacteria
rapid detection of
multi drug resistance (MDR)
Tuberculosis
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 • Beatrice Steinhöfel • Daniela Stephan • Tanja Struwe-Sonnenschein • Petra Vock • Birgit Voss
Ann -Kathrin Witt
lab strengthening, external quality assurance
MISSION
pulmonary tuberculosis between autochthonous and immigrant
The German National Reference Centre for Mycobacteria (NRC) is part of a worldwide laboratory network which aims to support and strengthen TB laboratories in high prevalence countries. The NRC is one of the 26 Supranational Reference Laboratories worldwide and since 2011 elected as representative of the SRL network.
sub-populations. BMC Infect Dis. 2009; 9:197.
65
internal and external collaborations
In house cooperation with the group of N. Reiling targets
the analysis of certain pharmaceutical compositions for the
clusters, defined as clusters including German and foreign-born
treatment of resistant strains. Cooperation with many national
most important findings
individuals, could be observed with increasing time spent by
organizations (RKI, Robert Koch Institute; public health offices;
Drug resistance in TB
immigrants in the host country.
GTZ, Gesellschaft für technische Zusammenarbeit; KFW,
Rapid detection of tuberculosis (TB) and resistant TB, mainly multi
External quality control and training
Kreditanstalt für Wiederaufbau; DAHW, Deutsche Lepra- und
drug resistant (MDR) and extensively drug resistant (XDR), is one
External quality control is getting a higher significance in
Tuberkulose-Hilfe; FLI, Friedrich Löffler-Institut Jena) are carried
of the main corner stones for prevention of further transmission
the laboratory diagnostics during the last years. The NRC is
out for lab strengthening purposes, for epidemiological studies,
of the disease. Molecular techniques enable the early detection
responsible for the external quality assurance (EQA) in the field of
Figure 1: Line Probe Assay for the rapid determination of resistance to flu-
and for analysis of NTM variability. Consultant activities, in order
of TB and resistance directly in patient specimens. Two newly
mycobacterial diagnostic, organized by INSTAND e.V. Düsseldorf. orochinolones, Amikacin, Capreomycin, and Ethambutol of M. tuberculosis
to contribute to necessary training activities to enhance human
strains
resources capacity for TB laboratory services, is provided for
developed molecular assays for the rapid detection of drug
This includes the preparation of samples for microscopy, primary
resistant TB have been evaluated. Within a multicenter,
isolation, identification of mycobacteria, and drug susceptibility
prospective evaluation of the Xpert MTB/RIF assay for detection
testing of M. tuberculosis. App. 250 national and international
selected puBlications
Frontières; ICRC, International Committee of Red Cross; USAID,
of TB and resistance to RMP, a high sensitivity and specificity in
laboratories are provided with samples twice a year. The results
Boehme CC, Nabeta P, Hillemann D, Nicol MP, Shenai S,
U.S. Agency for the International Development; FIND; The
detection of TB bacteria as well as detection of resistance to RMP
are analysed and certificates issued. Requests concerning
Krapp F, Allen J, Tahirli R, Blakemore R, Rustomjee R,
Foundation for Innovative New Diagnostics) e.g. for the set-up
in sputum specimens within two hours could be shown. The assay
quality control and diagnostic methods are discussed with the
Milovic A, Jones M, O’Brien SM, Persing DH, Ruesch-Gerdes
of TB laboratories and drug resistance surveillance in Armenia,
identified more than 97 % of all patients with culture-confirmed
laboratories on demand. Training in all fields of diagnostics is
S, Gotuzzo E, Rodrigues C, Alland D, Perkins MD. Rapid
Azerbaijan,
tuberculosis, including more than 90 % of patients with smear-
offered for the improvement of the laboratory procedures.
Molecular Detection of Tuberculosis and Rifampin Resistance. N
Moldova, Serbia, Turkmenistan, Uganda and Uzbekistan.
negative disease. Since resistance to RMP is high suggestive for
Non tuberculous mycobacteria
Engl J Med 2010 363:1005-1015.
MDR TB, this test enables the immediate identification of MDR TB
Non tuberculous mycobacteria (NTM) are isolated increasingly
within few hours. Furthermore, the rapid detection of XDR strains
from patient specimens due to improved diagnostic methods
Hillemann D, Rüsch-Gerdes S, Richter E. Feasibility of
for 32 European Countries for a European Center for Disease
could be proven by using a line probe assay for determination
and to rising implementation of immunosuppressive therapies.
the
Control (ECDC) project.
amikacin/capreomycin, and ethambutol resistance testing of
several international organizations (WHO; MSF, Médecins Sans
Bosnia-Herzogowina,
Kazakhstan,
Kyrgyzstan,
Furthermore, the NRC is work package leader in the EU TB PANNET, WP3: EQA for DR-TB and expansion, and accomplishes EQA
GenoType(R)
MTBDRsl
Assay
for
fluoroquinolone,
of resistance to fluorochinolones, Amikacin, Capreomycin, and
Definite identification of NTM species is eminently important
Ethambutol. Compared to conventional drug susceptibility
for final estimation of the clinical significance of different NTM
Mycobacterium tuberculosis strains and in clinical specimens. J
testing, fluoroquinolone resistance was detected in 8 (88.9 %) of
species and subspecies. Unambiguous identification of all
Clin Microbiol. 2009, 22.
9, Amikacin/Capreomycin resistance in 6/7 (75.0 %/87.5 %) of 8,
NTM species nowadays can only be performed by molecular
Grant support
RKI (National Reference Centre and network respiratory
infections), EU TB PAN-NET, ECDC: EQA
and Ethambutol resistance in 10 (38.5 %) of 26 resistant strains.
techniques, mostly by DNA sequence analysis. Furthermore,
Cox HS, Sibilia K, Feuerriegel S, Kalon S, Polonsky J, Khamraev
Epidemiological studies
the availability of molecular techniques enables the detection
AK, Rüsch-Gerdes S, Mills C, Niemann S. Emergence of
Several studies have been carried out to analyse the
of non growing mycobacteria, like M. genavense, mainly in
extensive drug resistance during treatment for multidrug-resistant
epidemiology of TB in the different settings in Germany and
immunosupressed patients. Attention has been focused on the
tuberculosis. N Engl J Med. 2008; 359:2398-400.
abroad by applying molecular strain typing techniques. Key
presence of NTM in animals as a possible source of infection
questions are the exploration of the population structure of M.
for immunosuppressed patients. M. genavense could repeatedly
Theuss T, Aupperle H, Eulenberger K, Schoon HA, Richter
tuberculosis complex strains in the different areas and possible
be shown in birds or other animals. Recently, new strain typing
E. Disseminated infection with Mycobacterium genavense in
interactions between metropolitan and rather rural settings. A
methods for M. avium isolates based on ‘multi locus sequence
a grizzled giant squirrel (Ratufa macroura) associated with
prospective study conducted with the respective public health
analysis’ (MLSA) has been proposed with the advantages that
the isolation of an unknown Mycobacterium. J Comp Pathol.
offices in Baden-Württemberg was finished recently. It was
it should be applicable to all M. avium subspecies. Applying
2010;143:195-8.
aimed to describe how immigration influences TB transmission
the technique to M. avium strains isolated from lymph nodes of
in Germany. Forty-seven clusters could be identified which
children may help to gain deeper insights into the epidemiology
Barniol J, Niemann S, Louis VR, Brodhun B, Dreweck C, Richter
included 132 (17 %) of the study participants. A trend to mixed
of cervical lymphadenitis.
E, Becher H, Haas W Junghanss T. Transmission dynamics of
Dept. Pneumology • National Reference Center for Mycobacteria
Dr. Sabine Rüsch-Gerdes
non tuberculous
mycobacteria
Molecular Mycobacteriology
Tuberculosis
molecular epidemiology
Dr. Susanne Homolka • Dr. Silke Feuerriegel • Dr. Andreas Roetzer • Tanja Struwe-Sonnenschein • Freda
Sührck • Tanja Ubben
evolution
resistance mechanisms
population structure
MISSION
which appears to be more prone to acquire resistance and
internal and external collaborations
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 and virulence of Mycobacterium tuberculosis complex (MTC) 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, host-pathogen interaction and virulence represents a highly challenging research topic.
to develop a hyper transmissible MDR phenotype. Currently,
Borstel TB Center
underlying genetic variation and mechanisms of virulence
Dag Harmsen Universität Münster; Jörn Kalinowski Universität
and resistance acquisition are unknown. We used massively
Bielefeld; Ulrich Nübel RKI Wernigerode; Sebastien Gagneux,
parallel whole-genome sequencing to compare a susceptible
Swiss TPH, Basel; Eman Aleksic, Burnet Institute, Melbourne;
strain to its MDR variant isolated from the same outbreak in
Kristin Kremer, Dick van Soolingen, National Institute of Public
Karakalpakstan (Uzbekistan), a region with a high rate of MDR
Health and Environment, Bilthoven;
tuberculosis. In addition to the backbone of Beijing specific
TBPANNET Consortium: 27 partner institutions in Europe and US.
EUMEDNETvsTB Consortium: 7 partner institutions in Europe and Africa.
most important findings
regarded as a highly homogeneous population; however,
variations present in both isolates (1,209 single-nucleotide-
Epidemiology of TB. Harmonized typing of bacteria and easy
recent data suggest the causative agents of tuberculosis are
polymorphisms), our analysis revealed an unexpectedly high
identification of locally or internationally circulating clones
more genetically and functionally diverse than appreciated
genomic diversity between the two isolates comprising 130
are essential for epidemiological surveillance and disease
previously. The impact of this natural variation on the virulence
single-nucleotide-polymorphisms and one large deletion
control. For MTC isolates, multi-locus variable number tandem
and clinical manifestations of the pathogen remains largely
(Fig. 1). Importantly, this diversity was not limited to the MDR
repeat analysis (MLVA) targeting mycobacterial interspersed
unknown. Therefore, we examined the effect of genetic diversity
variant. Our results suggest a much higher genomic diversity
repetitive units (MIRU) has been internationally adopted as
among MTC clinical isolates on global gene expression and
among M. tuberculosis outbreak clones in endemic settings as
the new standard typing method. However, no specialized
survival within macrophages. We discovered lineage-specific
previously anticipated. The high level of functional genomic
bioinformatics web tools are available for analyzing MLVA
transcription patterns in vitro and distinct intracellular growth
diversity demonstrates the extraordinary potential of TB strains
data. Therefore, we have developed the web application MIRU-
profiles associated with specific responses to host-derived
to adapt and might help to explain evolution of resistance
VNTRplus (http://www.miru-vntrplus.org). This freely accessible
environmental cues. Strain comparisons also facilitated
and virulence.
service allows users to analyze genotyping data of their
delineation of a core intracellular transcriptome, including
strains alone or in comparison with a reference database of
genes with highly conserved regulation across the global
strains representing the major MTC lineages. Analysis can be
panel of clinical isolates. This study affords new insights into
Comas I, Chakravartti J, Small PM, Galagan J, Niemann
based on MLVA-, spoligotype-, large sequence polymorphism
the genetic information that M. tuberculosis has conserved
S, Kremer K, Ernst JD, Gagneux S. Human T cell epitopes of
and single nucleotide polymorphisms (SNPs) data. Tools for
under selective pressure during its long-term interactions with
Mycobacterium tuberculosis are evolutionarily hyperconserved.
data exploration include search for similar strains, creation of
its human host.
Nat Genet. 2010 42(6):498-503.
Homolka S, Niemann S, Russell DG, Rohde KH. Functional
selected puBlications
phylogenetic trees and mapping of geographic information.
To facilitate scientific communication, an expanding genotype
Molecular determinants and microevolution of resistant
nomenclature has been implemented. During the last year,
strains. Resistance mechanisms of clinical M. tuberculosis
genetic diversity among Mycobacterium tuberculosis complex
the database had more than 4000 users distributed over 112
isolates have been investigated in several studies e.g. for first
clinical isolates: delineation of conserved core and lineage-
countries worldwide.
line (ethambutol) and second line drugs (para-aminosalicylic
specific transcriptomes during intracellular survival. PLoS
acid). Importantly, the data obtained demonstrate that particular
Pathog. 2010 6(7):e1000988.
The system also opens the door to multifunctional web based
SNPs are excellent marker for the emergence of resistance in
systems that combine automated strain identification with clinical
clinical isolates. On the other hand, we also found SNPs that
Intemann CD, Thye T, Niemann S, Browne EN, Amanua
and geographic information. These will be a breakthrough for
are specific for particular phylogenetic lineages such as Beijing
Chinbuah M, Enimil A, Gyapong J, Osei I, Owusu-Dabo
Figure 1: Overview of the genome data obtained (H37Rv, K-1, and K-2).
A) Venn diagram. B) SNP overview. C) Circular plot of H37Rv genome
and SNPs obtained.
Grant support
the detection of MDR-TB clusters and monitoring the international
or Haarlem. These data show that comparative genomics is a
E, Helm S, Rüsch-Gerdes S, Horstmann RD, Meyer CG.
spread of particular MDR clones and thus, will provide a
powerful tool to confirm suggested resistance mechanisms and
Autophagy gene variant IRGM -261T contributes to protection
BMBF: Network “Pulmonary Tuberculosis – Host and Pathogen
completely new tool for local and global TB surveillance.
to discover new once. However, phylogenetic markers represent
from tuberculosis caused by Mycobacterium tuberculosis but not
Determinants of Resistance and Disease Progression (TBornotTB)”.
potential confounders and MTBC population structure should
by M. africanum strains. PLoS Pathog. 2009 5(9):e1000577.
EU-FP7 Projekt (223681): Pan-European network for the study
Global
population
structure,
genomic
diversity
and
virulence. Elucidating the functional significance of MTC
be considered when defining new resistance mechanisms by
comparative sequencing.
genetic diversity is critical for understanding the diverse
and clinical management of drug resistant tuberculosis.
Weniger T, Krawczyk J, Supply P, Niemann S, Harmsen
EU-FP7 Projekt (245872): EUMEDNETvsTB Building a cooperative
D. MIRU-VNTRplus: a web tool for polyphasic genotyping of
strategy between Europe
outcomes of infection, treatment efficacy, and the development
To investigate the microevolution in MDR strains, we carried out
Mycobacterium tuberculosis complex bacteria. Nucleic Acids
and Mediterranean countries for upgrading tuberculosis
of new diagnostics and therapeutics. The MTC was initially
a genome investigation of clinical isolates of the Beijing lineage
Res. 2010 38:W326-31.
research and control.
67
Dept. Pneumology • Molecular Mycobacteriology
PD Dr. Stefan Niemann
Clinical and experimental pathology
HOPE technique
transcriptome /
proteome
Dr. Mahdi AbdullaH • Gabriele Cornehls • Stefanie Fox • Dr. Jürgen Galle • PD Dr. Torsten Goldmann •
Iris Jonas • Daniel Kähler • Angela Kelp • Maria Lammers • Dr. Dagmar Lang • Sebastian Marwitz •
Janine Radke • Heidi Scheibel • Dr. Holger Schultz • Dr. Florian Stellmacher • Gabriele Tänzer • Jasmin
Tiebach • Rolf Warnecke • Monika Wolgast • Andreas Zyzik
high throughput-validation
Prof. Dr. Dr. Ekkehard Vollmer
MISSION
bel P, Dalhoff K, Goldmann T. The TGF-beta-Pseudoreceptor
Networks: University of Tiflis, University of Baku, University of
One mission is to deliver state-of-the-art diagnostics focusing on diseases of the lung. Another area of interest is the development of new markers for so-called personalized medicine. Our central methodological tool is a functional human tissue culture model for different diseases of the lung. The base of this model is the HOPE-technique. For screening purposes and biomarker identification we apply transcriptome and proteome analyses, which are complemented by high throughput validation tools such as tissue microarrays.
BAMBI is strongly expressed in COPD lungs and regulated
Eriwan. Krankenhaus Großhansdorf (Ch. Kugler, Rabe K, Reck
by nontypeable Haemophilus influenzae. Resp Res 2010 May
M, H. Watz, Welker L)
31;11(1):67
Abdullah M, Schultz H, Kähler D, Branscheid D, Dalhoff K,
Figure 1:
Zabel P, Vollmer E, Goldmann T. Expression of the acute pha-
Immunohisto-
se protein haptoglobin in human lung cancer and tumor-free
chemical detec-
lung tissues. 2009 Pathol Res Pract 205(9):639-47
tion of BAMBI
most important findings
Western blot and immunohistochemistry (Figure 2) we showed
The most important finding during the past two years is the
the same on the protein level. Further studies are currently
Kähler D, Alexander C, Schultz H, Abdullah M, Branscheid
overexpressed
discovery of two immuno-regulatory molecules, which were
finished, which aim to elucidate the transcriptional regulation
D, Lindner B, Zabel P, Vollmer E, Goldmann T. Proteomics
not even known to be expressed in the human lung before.
and possible secretion as well as the immunologic functions
Out of the Archive: 2-D-electrophoresis and Mass Spectromet-
The first one is designated BAMBI (Bone morphogenetic pro-
of pHp. Due to the known immunomodulatory effects of hapto-
ry Using HOPE-fixed, Paraffin-embedded Tissues. J Histochem
tein and Activin Membrane-Bound Inhibitor) the second one is
globin, its broad expression and synthesis within human lung
Cytochem. 2010 Mar;58(3):221-28
called pulmonary haptoglobin (pHp). This was accomplished
tissues strongly suggests a function as a fundamental pulmo-
by application of the human STST-model (Short Term Stimulati-
nary local defense element.
in COPD (400x)
internal and external collaborations
Internal collaborations: NRZ(S. Rüsch-Gerdes, E. Richter, D.
on of Tissues); HOPE-fixation, transcriptome-analyses and high
throughput validation of the candidate genes. The third impor-
Proteomics: The crosslinking activity of formalin, which is used
Hillemann), Molecular Mycobacteriology (S. Niemann), Micro-
Figure 2:
tant finding was the development of a procedure to perform
as the standard fixative for tissues throughout the world, ham-
biology and Infectiology (U. Schaible), Microbial Inflammation
Expression
proteomics out of paraffin-embedded materials. Furthermore,
pers the application of many modern molecular read out sys-
Research (S. Ehlers, N. Reiling), Cellular Allergology (H.Haas),
of pHp in a
we implemented molecular diagnostics for the detection of
tems if compared to frozen materials. Frozen materials, on
Clinical and Molecular Allergology (A. Petersen, U. Jappe),
human lung
activating mutations of Ki-Ras and EGF-R, which are the prere-
the other hand, display a bad preservation of morphologic
Mucosa Immunology (A. Frey, K. Ramaker), Clinical Infectious
stimulated with
quisite for decision-making in targeted therapies.
details and are hard to store, which rules out such materi-
Diseases (C. Lange, U. Greinert, B. Karlsdorf), Inflammation
Pam3 (200x)
al as a standard in pathology. During the past ten years we
and Regenaration (H. Fehrenbach, Ch. Vock),Barrier Integrity (I.
have shown that the HOPE-technique is a useful alternative,
Lautenschläger), Pathophysiology of Inflammation (H.P.Hauber,
BAMBI: Based on transcriptome-data we identified BAMBI
as a candidate gene, which gets up-regulated upon ex vivo
since it preserves morphologic details as well as nucleic acids
K. Gaede), Cellular Pneumology (H. Haas), Medical and Bio-
infection with Haemophilus influenzae. We verified this data
and proteins. We have developed protocols for immunohis-
chemical Microbiology (H. Brade), Structural Biochemistry (O.
using RT-PCR. Under application of immunohistochemistry (Fi-
tochemistry, in situ hybridization, PCR, Northern-blot, RT-PCR,
Holst), Immunochemistry (U. Zähringer, C. Alexander), Biophy-
gure 1) and Western-blot we analyzed BAMBI on the protein
transcriptome-analyses, and Western-blot which all showed a
sics (T. Gutsmann, J. Andrä, K. Brandenburg, B. Lindner), Im-
level. BAMBI is a TGF-ß pseudo-receptor which can bind TGF-ß
superior quality of HOPE-fixed materials. Here we analyzed
munobiology (S. Bulfone-Paus, Z. Orinska), Biochemical Immu-
but does not give an intracellular signaling. The role of TGF-ß
the application of proteome-analyses to HOPE-fixed, paraffin-
nology (F. Petersen), Vet. Infection Biology and Immunology (J.
signaling in non-malignant diseases of the lung to date is of-
embedded tissues. We showed that such materials can be
Ahmed, U. Seitzer), Immunoepigenetics (P. Singh, J. Bullwinkel),
Grant support
ten controversially discussed; by the existence and function of
successfully subjected to 2-dimensional gel-electrophoresis (2-
Cellular Pneumology (C. Stamme), Innate Immunity (H. Heine),
BMBF project on the ex vivo infection of human lung tissues with
Fluorescence Cytometry (T. Scholzen)
Legionella (Legioprotect). Covance project on sexual transmis-
BAMBI we are now able to find explanations. BAMBI remains
DE). Since it is well known that there can be huge differences
under intense investigation in our group and other collabora-
between a proteome and a corresponding transcriptome, our
ting laboratories.
possibilities to apply 2-DE to archived tissues will substantially
External collaborations: University of Lübeck, Med Clinic III (K.
role of TKTL-1 in Non Small Cell Lung Cancer. Roche project on
pHp: Also identified by transcriptome analyses we performed
promote the knowledge of what’s going on in the course of a
Dalhoff, D. Drömann). University of Lübeck, Medical Microbiolo-
the development of a procedure to acquire broncho-alveolar
a broad study on pHp. Haptoglobin is a long known molecule,
disease in the human system
gy (J. Rupp). University of Dohuk, Iraq (M. Abdullah). University
–lavages and sputum with the HOPE-technique. DAAD projects
of Freiburg, Pneumology (J. Müller-Quernheim, G. Zissel, A. Pras-
supporting the medical development of the southern Caucasus.
which was thought to be synthesized in the liver. There was no
sion of a gene-therapeutic vector. r-biopharm project on the
data on the possible synthesis of a pulmonary haptoglobin
selected puBlications
se). Charite, Pneumology (S. Hippenstiel, B. Schmeck). BMBF
DGP project on immunomodulators in expanded lungs. BMBF
in the human system. By means of RT-PCR and RNA in situ
Dromann D, Rupp J, Rohmann K, Osbahr S, Ulmer AJ, Mar-
Medizinische Infektionsgenomik: University of Braunschweig, Ro-
project for development of new diagnostic markers. Astra-Ze-
hybridization we showed that pHp exists and is indeed synthe-
witz S, Roschmann K, Abdullah M, Schultz H, Vollmer E, Za-
bert Koch Institute Wernigerode University of Greifswald. DAAD
neca: Iressa study on NSCLC
Dept. Pneumology • clinical and
xxxxxxxxxxxxxxxxxxxxx
experimental pathology
sized in high levels within the human lung. By application of
69
Cellular Allergology
Immunomodulation
Daniela Barths • Birte Hanisch • Dr. Gabriele Schramm
by
parasitic worms
PD Dr. Helmut Haas
MISSION
H, Doenhoff M. Molecular characterisation of kappa-5, a
Grant support
To identify and characterize anti-inflammatory molecules from parasitic worms, namely Schistosoma mansoni, and to employ them for protection against allergies and autoimmune disorders. Our motto:
“Learn from the Worm”
major antigenic glycoprotein from Schistosoma mansoni eggs.
BMBF 0315277 „In vitro culture of Schistosoma mansoni – Culture
MOLECULAR AND BIOCHEMICAL PARASITOLOGY 2009; 166:4-14.
dish instead of mammalian host“, September 2008 – August
EU-FP7, grant agreement no. 242107 „The targeted development
the particular IgE-inducing capacity of Kappa-5 reflects the
Dieckmann C, Haas H, Mueller-Dieckmann J. Cloning,
of a new generation Vaccine for Schistosomiasis” (TheSchistoVac;
Omega-1, a major secretory Schistosoma mansoni egg
fact that this molecule, in contrast to the other two major egg
expression, purification, crystallization and preliminary X-ray
work package 5), March 2010 – February 2014, 1 Post-Doc
antigen, drives Th2 responses
antigens, is not secreted and, thus, differently processed or
crystallographic analysis of interleukin-4-inducing principle
Ovamed GmbH, Barsbüttel, Germany, November 2009 –
S. mansoni soluble egg antigens (SEA) induce strong Th2
whether it acts as a “genuine worm allergen” remains to be
from
responses both in vitro and in vivo. However, the specific
established.
CRYSTALLOGRAPHICA SECTION F-STRUCTURAL BIOLOGY AND
Schistosoma
mansoni
eggs
(IPSE/alpha-1).
ACTA
CRYSTALLIZATION 2009; 65:594-596
have not been identified. In collaboration with other research
groups, we found that Omega-1, a major glycoprotein
Waknine-Grinberg J, Gold D, Ohayon A, Flescher E, Heyfets
secreted from S. mansoni eggs and present in SEA, is capable
A, Doenhoff M, Schramm G, Haas H, Golenser J. Schistosoma
of conditioning human monocyte-derived dendritic cells in vitro
mansoni infection reduces the incidence of murine cerebral
to drive T helper 2 (Th2) polarization. Furthermore, using IL-4
malaria. MALARIA JOURNAL 2010; 12:881-888.
dual reporter mice, both natural and recombinant Omega-1
alone were sufficient to generate Th2 responses in vivo, even
internal and external collaborations
in the absence of IL-4R signaling. Finally, Omega-1-depleted
Internal:
SEA displayed an impaired capacity for Th2 priming in
Thomas Scholzen
vitro, but not in vivo, suggesting the existence of additional
Guntram Grassl
factors within SEA that can compensate for the Omega-1-
Martin Ernst
mediated effects. These results were supported by similar and
T. Goldmann
complementary findings obtained by Steinfelder et al. and
Holger Heine
were published as companion papers. Collectively, Omega-1,
Norbert Reiling
Zane Orinska
a single component of SEA, was identified as a potent inducer
of Th2 responses.
Figure 1: In vitro culture of Schistosoma mansoni. Supported by BMBF
Arnd Petersen/Uta Jappe
(grant 0315277), in our lab an in vitro system for propagating schistoso-
Kappa-5, a major S. mansoni egg antigen, is a preferential
me larvae (schistosomula) in the presence of blood components and
External:
target of IgE
growth factors has been established. Within 6 weeks of culture, the
Maria Yazdanbakhsh, C.H. Hokke, Leiden University Medical
We have isolated, cloned and characterized Kappa-5, one of
larvae mature into adult parasites including pair formation.
Center, Leiden, NL
the three major glycoproteins produced by S. mansoni eggs.
Dragana Jankovic, National Institute of Allergy and Infectious
These proteins (Omega-1, IPSE/alpha-1 and Kappa-5) have
selected puBlications
Diseases (NAID),
earlier been found to be primary targets of the egg-directed
Everts B, Perona-Wright G, Smits H, Hokke C, van der Ham
Bethesda, USA
antibody response of the host. No significant similarities
A, Fitzsimmons C, Doenhoff M, van der Bosch J, Mohrs K,
M. Sattler, Technische Universität, München
of Kappa-5 to known sequences from species other than
Haas H, Mohrs M, Yazdanbakhsh M, Schramm G. Omega-1,
J. Müller-Dieckmann, EMBL, Hamburg
schistosomes were found. While IPSE/alpha-1 and Omega-1
a glycoprotein secreted by Schistosoma mansoni eggs, drives
C.G. Grevelding, Universität, Giessen
are expressed only in eggs, Kappa-5 is present in eggs as
Th2 responses. JOURNAL OF EXPERIMENTAL MEDICINE 2009;
M. J. Doenhoff, University of Nottingham, UK
well as in miracidia at mRNA and protein level. Interestingly,
206:1673-1680.
A. Cooke, University of Cambridge, Cambridge, UK
IPSE/alpha-1- and Omega-1-specific antibodies from sera of S.
D.W. Dunne, University of Cambridge, UK
mansoni-infected Africans belonged to the IgG isotype only,
Schramm G, Hamilton J, Balog C, Wuhrer M, Gronow A,
F.H. Falcone, University of Nottingham, UK
while the antibodies to Kappa-5 were both of the IgG and –
Beckmann S, Wippersteg V, Grevelding C, Goldmann T,
J. Golenser, Hadassah Medical School, Jerusalem, IL
in over 50 % of individuals – also of the IgE isotype. Whether
Weber E, Brattig N, Deelder A, Dunne D, Hokke C, Haas
M. Stadecker, Tufts University School of Medicine, Boston, USA
September 2010, 1 TA
Dept. Pneumology • Cellular Allergology
2011, 1 post-doc, 1 technician
Mayerhofer H, Schramm G, Hatzopoulos G, Mueller-
most important findings
molecules that prime the development of Th2 responses
71
Clinical and Molecular Allergology
from clinic
to research
Marisa Böttger • Gerda Finnern • Markus Hartmann • Saskia Hellmig • Maren Hohn • Prof. Dr. Arnd
Petersen • Sandra Rennert • Simone Ross • Dr. Frauke Schocker • Daniela Warneke
and back to patient
Prof. Dr. Uta Jappe
supported this notion in more detail. After establishing a master
• 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
gel, 110 timothy grass pollen specific protein spots identified by
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. Jappe
• Prof. Dr. T. Gutsmann: Functional microbiological studies on
peanut defensin
mass spectrometry were annotated. All 10 groups of grass pollen
National
allergens were detected. Mostly, several isoforms were found,
• Prof. Dr. J. Saloga (University of Mainz): Modulation of the
especially of Phl p 1, 5, and 13. Qualitative and quantitative
human immune response using allergenic proteins, DNA,
differences were determined at the level of individual allergens.
and adjuvans on APCs
While two of the extracts varied quantitatively concerning their
components of +/-5fold, the comparison with a third extract
• Dr. S. Schillberg (Fraunhofer Institute, Aachen): Expression of
allergens in tobacco
revealed differences of up to +/-60fold. Some isoforms were even
• Prof. Dr. C. Traidl-Hoffmann (ZAUM, Fraunhofer Institute, Munich):
completely missing. In addition, Phl p 13 showed a variation
allergen uptake via human skin and subsequent processing
in molecular masses indicating protein degradation. Although
• Prof. Dr. O. Janßen (Institute for Immunology, UK SH, Campus
most important findings
difference gel electrophoresis (2D-DIGE)
allergen extracts are much more difficult to standardize than
Kiel): Proteomic approach (2D DIGE and mass fingerprinting)
Evidence for specific oil-allergen interactions
Allergen extracts used for allergy diagnosis and therapy are not
recombinant allergens, the proteins are correctly folded, bear
for the analysis of pollen and food allergens
Of the eleven identified peanut allergens (Fig. 1), Ara h 8 is
well characterized concerning their single constituents. Increasing
the authentic posttranslational modifications and their production
an important one in the oil plant peanut which affects patients
demands of the detailed knowledge of their components and
is less expensive. The described technique allows the qualitative
with combined birch pollen and peanut allergy. It is a Bet v
their composition, however, is necessary to determine the
and quantitative comparison of allergen extracts to the level
International
• Prof. Dr. J. Schröder (Institute for Dermatology, UK SH,
Campus Kiel): Defensins Isolation and characterization
1-homologous protein that can cause severe allergic reactions.
strategies leading to an optimal therapeutic outcome. With regard
of single components, and seems to be a valuable tool to
• Dr. C. van Drunen (ENT Department, Academic Medical
Recent studies showed that the structure of the major birch
to its respective suitability, a multiplex proteomic technique to
standardize complex allergen extracts according to their single
Center, Amsterdam): Stimulation of the innate immune
pollen allergen Bet v 1 forms a hydrophobic pocket which can
simultaneously determine the quality and quantity of the individual
components.
bind lipids. We analyzed Ara h 8 for its hydrophobic binding
components in allergenic grass pollen extracts was investigated.
system by grass pollen allergens
• Dr. A. Sette (La Jolla Institute for Allergy and Immunology,
properties, determined putative amphiphilic ligands and studied
Timothy grass pollen (Phleum pratense) were obtained from
selected puBlications
San Diego, USA): Molecular determinants of T cell epitope
the influence of lipid binding on the digestibility of the allergen.
three different suppliers, extracted under similar conditions
Krause S, Reese G, Randow S, Zennaro D, Quaratino D,
recognition
Displacement assays provided evidence for a hydrophobic cavity
and investigated by 2D-DIGE. Quantitative comparison of the
Palazzo P, Ciardiello MA, Petersen A, Becker WM, Mari A.
in the Ara h 8 molecule. Further investigations with putative ligands
individual protein spots was determined using CyDye staining. 2D
The lipid transfer protein (Ara h 9) as a new peanut allergen
Identification of food allergens considering geographical
showed strong lipid interactions especially with fatty acids.
immunoblotting was performed using sera of grass pollen allergic
relevant for a Mediterranean allergic population. J ALLERGY
aspects
• Dr. A. Mari, Center of Mol. Allergology, IDI-IRCCS, Rome:
• Prof. Dr. T.M. Platts-Mills (University of Virginia, Health
Experiments with nAra h 8 confirmed a genuine lipid binding.
Figure 1:
Gastrointestinal digestion experiments with rAra h 8 (without
Identification of
lipids) showed a rapid degradation by pepsin and gastric and
further peanut
Schmidt H, Gelhaus C, Nebendahl M, Janssen O, Petersen A.
pancreatic surrogates, whereas nAra h 8 (with lipids) remained
allergens by
Characterization of Phleum pratense pollen extracts by 2-D DIGE
inert towards gastric digestion. Ara h 8 binds lipids in a specific
2D DIGE tech-
and allergen immunoreactivity. PROTEOMICS 2010; 10:4352-4362.
Grant support
way, which stabilizes the molecule against gastrointestinal
nique (Schmidt
digestion. This causes a prolonged retention time/exposure in
et al. 2010)
Jappe U, Vieths S. Lupine, a source of new as well as hidden
responses of the lung“, Project Z1: Core facility for the mass
CLIN. IMMUNOL. 2009;124:771-778.
System, Charlottesville, USA): cross-reactive carbohydrate
determinants (alpha-GAL)
Special research field (SFB)/Transregio 22: „Allergic immune
the gastrointestinal tract, which might increase sensitization and
food allergens. MOL. NUTR. FOOD RES. 2010;54:113-126.
result in stronger allergic reactions. As a further result of the Ara
spectrometric analysis of allergy-relevant biomolecules and for
the characterization of natural and recombinant aeroallergens
h 8 purification, we isolated a protein, which does not seem to
internal and external collaborations
be an allergen, but is a defensin with activity against several
Inhouse
clinically relevant bacteria. This might be of therapeutic relevance
• PD Dr. A. Frey: Processing of food allergens, epitope mapping
when using peanut oil as ingredient in dermatological ointments
patients and monoclonal antibodies and polyclonal antisera
for atopic patients and opens strategies for investigations with
directed against grass pollen allergens. Identification of the
regard to allergy, atopy and infection.
protein spots and isoforms was done by mass spectrometry. 1D
• Prof. Dr. H. Fehrenbach: mouse models for asthma induction
• PD Dr. T. Goldmann: Interactions between allergens and
membranes
Western blotting pointed to differences in the protein patterns of
• PD Dr. H. Heine: Analysis of mediators liberated in allergy
Multiplex analysis: Comparison of grass pollen extracts
Phl p 5 and Phl p 13 by use of monoclonal antibodies. Quantitative
• PD Dr. B. Lindner: Characterization of allergens by mass
used for diagnosis and therapy by two-dimensional
comparison
performing
2D-DIGE
and
2D-immunoblotting
spectrometry
73
Dept. Pneumology • Clinical and Molecular Allergology
MISSION
Mucosa Immunology
Molecular Imaging
Mucosal Barriers
Dr. Barbara Frey • Özge Kök • Dr. Thorsten Krause • Iryna Kruse • Dr. Pia Müller • Verena Puls • Dr. Katrin Ramaker • Dr. Niels Röckendorf • Alheidis von Quast • Imke Wysokinski
High Throughput
Peptide Assays
Molecular Evolution
MISSION
In an additional part of our project we want to elucidate if
Inst. of Biomedical Optics, University of Lübeck; Inst. of Physical
The Division of Mucosal Immunology has devoted itself to the understanding, diagnosis, prevention and treatment of inflammatory, neoplastic and allergic conditions at the mucosal surfaces of the respiratory system and the gastrointestinal tract.
intestinal modifications of ingested materials may also be of
Chemistry, University of Hamburg; MPI for Polymer Research,
relevance in the development of food allergies. Food allergies
Mainz
xenobiotics at mucosal surfaces. As the digestive tract is one
are immune defense reactions against certain food proteins
BMBF-funded research consortium OPTOPROBE: ATTO-TEC,
and may be caused by a lack of mucosal tolerance against the
Siegen; University of Applied Sciences, Paderborn; GeSiM,
respective protein. A faulty tolerance induction could be due to
Dresden; Karl Storz, Tuttlingen; LMTB, Berlin; R-Biopharm,
Darmstadt
most important findings
of the key sites at which the body encounters foreign matter
premature destruction or incomplete formation of tolerogenic T
The research activities of the Division of Mucosal Immunology
we investigate the interactions of different kinds of ingested
cell epitopes in the gut. To test this hypothesis, we determine
SPP “Biomedizintechnik”: Inst. of Medical Engineering and Dept.
can be divided into two key projects:
material, e.g. food constituents, drugs, antigens or particulate
the stability of potential T cell epitopes in the intestine using our
of Otolaryngology, University of Lübeck
The “Molecular Imaging Project” deals with the diagnosis of
matter, with the different components of the small intestinal
peptide degradability assay. In this set-up, the half-lifes of 6400
Dept. of Organic Chemistry, University of Kiel
pathological conditions at mucosal surfaces. We want to create
mucosa, identify pathways along which this material enters the
10mer and16mer peptides derived from 18 major food allergens
Veterinary Research Institute, Brno, Czech Republic
smart contrast agents and probes for in-vivo molecular imaging
mucosa and determine the ensuing metabolic reactions and
and 5 non allergenic control proteins in murine intestinal fluid
and in-vitro point of care diagnostics. Targeted disease areas
immunological responses. With this approach we hope to find
ranged from 52 µs to about 10 min. We now have to check
include cancers of the colon and the lung and inflammatory
out which events and properties of ingested material govern
if comparably stable allergen-derived fragments constitute
pulmonary diseases such as COPD and asthma.
its classification towards the induction of immunologic defence,
potential tolerogenic T cell epitopes. To do so we have devised
The diagnostic power of most imaging modalities can be
tolerance or nonresponsiveness, and how misclassification
a strategy to screen peptide libraries derived from defined food
enhanced by employing specific contrast agents that enable
may lead to pathologic conditions such as allergies and
allergens for binding of MHC class II molecules and to submit the
the visualization of a disease marker down to the (sub)cellular
inflammations.
respective MHC II-peptide-complexes to a T cell binding assay.
level. In our approach to develop such contrast agents, we
In the course of our investigations we found that for predicting
want to combine a reporter unit, e.g. a fluorophore or an MRI
the fate of taken-up material it is essential to first analyze the
selected puBlications
label, with a peptide-based targeting ligand. In order to identify
modifications ingested compounds undergo upon contact with
Bade S, Gorris HH, Koelling S, Olivier V, Reuter F, Zabel
candidates that may be suited as ligands we devised a strategy
the preepithelial environment. This applies particularly to the
P, Frey A. Quantitation of major protein constituents of murine
for a directed “molecular evolution” of peptides with defined
action of the proteolytic enzymes in the digestive tract as well
intestinal fluid. ANALYTICAL BIOCHEMISTRY. 2010; 406:157-165.
properties. The process starts with the synthesis of a population
as to the impact of adsorbents such as mucus. We therefore
of lead peptides which then are screened for the desired
established procedures to harvest and standardize undiluted
Gorris HH, Bade S, Röckendorf N, Albers E, Schmidt MA,
properties, ranked accordingly and crossed and mutated using
intestinal fluid and mucus from various sources and developed
Franek M, Frey A. Rapid profiling of peptide stability in
Figure 2:
a genetic algorithm to create a filial generation. With this joint
assays and reagents for analyzing the degradability of peptides
proteolytic environments. ANALYTICAL CHEMISTRY. 2009;
Potential fate of
in silico/in vitro optimization method stepwise improvement of
and proteins and for the sensitive detection of biomolecules.
81:1580-1586.
ingested parti-
Figure 1:
Delivery of
fluorophore
payload into
the endosomes
of cultured cells
with the aid
of membrane
penetrating
peptides;
bar = 10 µm
culate material
the peptides is achieved until a ligand is obtained that meets
We now use these tools to unravel the processes that ingested
the requirements for the application of interest. In cooperation
materials face at the mucosae.
Bade S, Röckendorf N, Franek M, Gorris HH, Lindner
at the intestinal
with a network of academic and industrial partners we have
One focus of our investigations are nanoparticles which, due to
B, Olivier V, Schaper K-J, Frey A. Biolabeling with
epithelium: coa-
developed a robotic platform for the automated synthesis and
the expanding field of nanotechnology, are quite omnipresent
2,4-dichlorophenoxyacetic acid derivatives: the 2,4-D tag.
ting, aggregati-
screening of peptides and have set-up a variety of assays to
and may easily be taken up and incorporated inadvertently. In
ANALYTICAL CHEMISTRY. 2009; 81:9695-9702.
on, entrapment,
evaluate distinct properties of the peptides, e.g. affinity to a
order to clarify the physiological events and potentially harmful
target, stability in proteolytic environments or capability to direct
consequences connected with such uptake (Figure 2), we study
internal and external collaborations
uptake &
a payload to intracellular targets (Figure 1). By ultrastructural
the biochemical properties, cell binding, transepithelial transport
Inhouse:
transport
analysis of human colon cancer tissue we could identify a tumor
and tissue distribution of nanoparticles. When analyzing the
Divisions of:
marker for which we have designed a contrast agent composed
interaction of nanoparticles with intestinal fluid and mucus we
Barrier Integrity, Biophysics, Clinical and Experimental Biology,
of a peptidic ligand and a particulate reporter unit. This novel
found that, depending on their surface properties, nanoparticles
Clinical and Molecular Allergology, Pathophysiology of
contrast agent can now be validated in an animal tumor model
will more or less be coated with digestive tract contents which
Inflammation
which is currently being established.
in turn can lead to the formation of µm-sized agglomerates. In
The “Mucosal Interface Project” explores the processing,
consequence of this, nanoparticle binding to and/or uptake by
External:
Deutsche Forschungsgemeinschaft (DFG), grant Fr 958/5-1
transport and presentation of allergens, immunogens and
intestinal epithelial cells is strongly reduced.
DFG-funded research cluster PARENTRY: Inst. of Anatomy and
University of Lübeck, grant F249607
degradation,
Grant support
Bundesministerium für Bildung und Forschung (BMBF), grant
13N10505
75
Dept. Pneumology • Mucosa Immunology
PD Dr. Andreas Frey
Clinical Infectious Diseases
clinical
counseling
diagnosis
Prof. Dr. Christoph Lange
Kalsdorf • Lenka Krabbe • Benjamin Krummel • Leonhard Leid • Johannes Ortmann • Dr. Alan Strassburg
Janine Wolske
MDR-TB
tuberculosis
MISSION
by previous history of tuberculosis. BAL ELISpot is an important
hard Walzl, University of Stellenbosch, Stellenbosch (R. South
Our mission is excellence in tuberculosis (TB) and NTM-infections care, research, training and counselling. We are interested in evaluating new diagnostic and treatment strategies for susceptible and drug resistant TB, individualized patient care, the investigation on risk factors and outcomes in MDR- and XDR- TB, in improving TB prevention by investigation of fundamental mechanisms of the immune responses against Mycobacterium tuberculosis (Mtb), including HIV-coinfection, and in sustained capacity building in high burden countries to support the global fight against TB.
advancement to rapidly distinguish sputum AFB smear-negative
Africa); Robert Wilkinson, University of Cape Town (R. South Af-
tuberculosis from LTBI in routine clinical practice.
rica); and multiple European partners in TBNET studies and the
Pan-European MDR-TB database project.
selected puBlications
Leidl L, Mayanja-Kizza H, Sotgiu G, Baseke J, Ernst M, Hirsch
Figure 1:
C, Goletti D, Toossi Z, Lange C. Relationship of immunodiag-
International
nostic assays for tuberculosis and numbers of circulating CD4+
collaborations
T-cells in HIV–infection. Eur Respir J 2010, 35:619-26
on clinical TB
most important findings
pot by 40.8, 19.3, and 2.5 times, respectively. Individuals with a
1. Identification of latent infection with Mtb in HIV-infected
positive IFN-γ ELISpot/negative IL-2 ELISpot result had a median
individuals: Infection with the HIV is the greatest risk factor for
(IQR) duration of index case exposure of 568 hours (133_1000)
Krummel B, Strassburg A, Ernst M, Reiling N, Eker B, Rath H,
de partners in
TB in Africa. Tuberculin-skin-test (TST), QuantiFERON-TB Gold In-
compared to individuals with a positive IFN-γ ELISpot/positive
Hoerster R, Wappler W, Schoellhorn V, Glaewe A, Sotgiu G,
the Republic of
Tube (QFT-G-IT) and T-Spot.TB assays were performed in newly
IL-2 ELISpot result (median=24 hours; 20_130; p-value= 0.047).
Lange C. Potential role of IL-2 ELISPOT in differentiating recent
Moldova, the
diagnosed HIV-infected individuals with and without active TB
Combination of a Mtb specific IFN-γ ELISpot with a Mtb-specific
and remote infection in tuberculosis contact tracing. PLoS One
country with the
and in HIV-uninfected persons at an University outpatient clinic
IL-2 ELISpot significantly improved the identification of individu-
2010; 5(7):e11670
highest preva-
in Kampala, Uganda. A total of 135 individuals were enrolled:
als with the highest risk of recent Mtb infection and is a pro-
109 with a new diagnosis of HIV-1 infection, but no active TB, 19
mising method that should be explored to target tuberculosis
Jafari C, Thijsen S, Sotgiu S, Goletti D, Domínguez Benítez
with HIV-1 infection and active TB, and 7 HIV-uninfected healthy
preventive chemotherapy.
JA, Losi M, Eberhardt R, Kirsten D, Kalsdorf B, Bossink A,
research inclu-
lence MDR-TB
in Europe.
Latorre I, Migliori GB, Strassburg A, Winteroll S, Ulf Greinert
subjects. In control subjects immune responses were positive in
57.2 % by TST and in 100% by at least one IGRA. In HIV-1 infected
3. Improving the rapid detection of AFB smear negative tu-
U, Richeldi L, Ernst M and Lange C for the TBNET. Bronchoal-
Figure 2:
patients without active TB, induration in the TST (mm) (rho=0.41,
berculosis: The rapid diagnosis of pulmonary tuberculosis is
veolar lavage enzyme-linked immunospot for a rapid diagnosis
A novel method
p-value<0.0001) and concentration of IFN-γ in the QFT-G-IT tube
difficult when acid fast bacilli (AFB) cannot be detected in spu-
of tuberculosis: a TBNET Study. Am J Respir Crit Care Med 2009,
for the rapid
with MTB-specific antigens (rho=0.38; p-value: 0.0001) were
tum smears. Following a proof of principle study we examined
180:666-73
detection of
negatively correlated to numbers of circulating CD4+ T-cells
in routine clinical practice whether individuals with sputum AFB
(cells/µl) while numbers of IFN-γ producing cells (rho=0.03-0.13;
smear-negative tuberculosis can be discriminated from those
internal and external collaborations
p-value:0.21-0.77) and frequencies of positive test results for T-
with latent tuberculosis infection by local immunodiagnosis
Internal collaborations with Martin Ernst, Immunecell-Analytics;
Spot.TB test among groups of patients with different levels of im-
with a Mtb-specific enzyme-linked immunospot assay (ELISpot;
Stefan Ehlers, Molecular Inflammation Medicine; Holger Hei-

77
sputum AFB
munodeficiency remained constant (p=0.46). In HIV-1 infection,
T-SPOT .TB). Suspects of active tuberculosis unable to produce
ne, Innate immunity; Christoph Hölscher, Infection-Immunology;
TST and QFT-G-IT immune responses are both strongly related
sputum or with AFB negative sputum smears were prospectively
Stefan Niemann, Molecular Mycobacteriology; Norbert Reiling,
to the degree of immunodeficiency, while results of the T-Spot.TB
enrolled at TBNET centers in Europe. ELISpot with ESAT-6 and
Microbial Interface Biology; and Sabine Rüsch-Gerdes, Myco-
are independent of the level of CD4 T-cell depletion.
CFP-10 peptides was performed on peripheral blood mononu-
bacteriology.
smear-negative
pulmonary
tuberculosis by
enumeration of
antigen specific
T-cells at the
site of the
infection.
clear cells (PBMCs) and bronchoalveolar lavage mononuclear
2. A new method to improve tuberculosis contact tracing:
cells (BALMCs). Mtb-specific nucleic acid amplification (NAAT)
External collaborations with Jan Andersson, Karolinska Institute,
IGRA have improved tuberculosis contact tracing, but discrimi-
was performed on BAL fluid. Seventy-one of 347 (20.4 %) pa-
Stockholm (Sweden); Victor Botnaru, Institute for Phtisiopneu-
nation of recent from remote Mtb contacts is not possible by
tients had active TB. Out of 276 patients with an alternative di-
mology „Chriril Draganiuc“, Chisinau (Modova); Delia Goletti,
Supported by the Excellence Cluster „Inflammation at Inter-
IGRA alone. We present results of a tuberculosis contact investi-
agnosis 127 (46.0 %) were considered to be latently infected with
National Institute for Infectious Diseases, La Spalanzzani, Rome
faces“, European Union (EU-FP7), European Respiratory Socie-
Grant support
gation with a new early-secretory antigenic-target (ESAT)-6 and
Mtb by a positive PBMC ELISpot result. Sensitivity and specifi-
(Italy); Michael Hölscher, Ludwig- Maximilians University, Mu-
ty (ERS), HW & J Hector Foundation, Tuberculosis Network Eu-
culture-filtrate-protein (CFP)-10 specific interleukin (IL)-2 ELISpot
city of BALMC ELISpot for the diagnosis of active pulmonary TB
nich (Germany); Harriet Mayanja-Kizza, Makerere University,
ropean Trialsgroup (TBNET), Federal Ministry for Science and
in addition to ESAT-6 and CFP-10 specific IFN-γ ELISpot and TST.
were 91 % and 79 %, respectively. The BALMC ELISpot (OR 40.4)
Kampala (Uganda); Dick Menzies, McGill University, Montreal
Technology (BMBF).
Results of the TST, IFN-γ ELISpot and IL-2 ELISpot were positive in
was superior to PBMC ELISpot (OR 10.0), tuberculin skin test
(Canada); Giovanni B. Migliori, Fondazione Maugeri, Tradate
6/172 (3.4 %), 7/167 (4.2 %) and 6/196 (3.1 %) of contacts, respec-
(OR 7.8) and Mtb-specific NAAT (OR 12.4) to diagnose sputum
(Italy); Martina Sester, Saarland University, Homburg (Germa-
tively. Close contact (≥ 100 hours) to the index case increased
AFB smear-negative tuberculosis. In contrast to PBMC ELISpot
ny); Giovanni Sotgiu, University of Sassari, Sassari (Italy); Zahra
the risk of positive results in the IFN-γ ELISpot, TST, and IL-2 ELIS-
and tuberculin skin test, the BALMC ELISpot was not influenced
Toossi, Case Western Reserve University, Cleveland (USA); Ger-
Dept. Pneumology • Clinical Infectious Diseases
IGRA
Franziska Daduna • Dr. Christiane Gerlach • Andrea Glaewe • Dr. Gunar Günther (deputy) • Dr. Ulf
Greinert • Tim Grüber • Christian Gutsfeld • Dr. Christian Herzmann • Dr. Claudia Jafari • Dr. Barbara
Impressum
Academic degree and professional qualification 2009/2010
total budget 2009
24.089 j
institutional funding ( federal + federal state)
16.940 j
third-party funding
7.149 j
of which DFG
2009
2010
11
5
Diploma
4
5
Bachelor of Science
3
2
Dissertation
29 %
3.798 j
53 %
1.335 j
19 %
EU project funding
757 j
11 %
Technicians
industry
887 j
12 %
Animal Keeper
foundations
254 j
4 %
miscellaneous
118 j
2 %
federal/federal state
Master of Science
1
10
11
1
1
2009
2010
internal
19
9
external
8
29
2009
2010
national
24
35
international
55
98
Conferences and Workshops 2009/2010
total budget 2010
27.122 j
institutional funding (federal + federal state)
17.310 j
third-party funding
9.812 j
36 %
Guest scientists 2009/2010
of which DFG
2.664 j
27 %
federal + federal state
3.069 j
31 %
EU project funding
2.605 j
25 %
industry
1.006 j
10 %
foundations
313 j
3 %
miscellaneous
155 j
1 %
Peer review publications 2009/2010
2009
153
2010
190
Patents and Licenses 2009/2010
Books and book articles 2009/2010
Assignation of patents
1
Stock of patent families
16
2009
11
4
2010
28
Stock of licensing agreements
Income from royalties (T j)
325.000 (2009)
477.993 (2010)
79
Facts & Figures
Funding (in thousands)
Impressum
International Networks 2009/2010
German Research Foundation ( DFG) SFB/TR 22, Allergische Immunantwort der Lunge
Administrative and Technical Services
6th and 7th EU-Frame Program
ICTTD: Integrated Consortium on Ticks and Tick-borne Diseases
Board of Directors
SFB 415, Spezifität und Pathophysiologie von Signaltransduktion-
GABRIEL : A multidisciplinary study to identify the genetic and
Prof. Dr. Peter Zabel (Acting Director)
swegen environmental causes of asthma in the European Community Prof. Dr. Ulrich Schaible
SFB 617, Molekulare Mechanismen der epithelialen Abwehr GALtrain (Marie Curie EST): Exposure to micro-organisms and
Prof. Silvia Bulfone-Paus
SFB 654, Plastizität und Schlaf suppression of allergic diseases: A European research training
SPP 1313, Biological Responses to Nanoscale Particles
scheme Administration
SPP 1394, Mast cells - promotors of health and modulators of
ASEMDialog: The EU, China and South East Asia dialog for the
Susann Schrader (Head)
disease development of research areas in animal health of mutual inter-
Sandra Groth (Hospital Management)
KFO 170, Frühpathogenese der Wegnerschen Granulomatose
est [coordination, FZB]
Dirk Grünes (Technical Building and Construction Manage-
FOR 585, Pathogen-spezifische Abwehrmechanismen in der
ConFluTech: Capacity building for the control of Avian influenza
ment)
Milchdrüse
through technology transfer and training [coordination, FZB]
Karl-Heinz Lemm (Staff Management)
TM-REST: A new platform for fast molecular detection of MDR
Sibylle Lenkeit (Cash Management)
rica/DFG “Development and validation of a sensitive LAMP as-
and XDR resistant strains of M. tuberculosis and of drug resist-
Heinz-Jürgen Wefel (Infrastructural Management)
say for the specific detection of Theileria sp.(sable) in sable
ant malaria
Joint research grant National Research Foundation South Af-
(Hippotragus niger) and roan (Hippotragus equinus) antelope”
Arbo-Zoonet: International network for capacity building for the
Computer Services
Deutsch-Afrikanische Kooperationsprojekte in der Infektiologie
control of emerging viral vector borne zoonotic diseases
Michael Reinhold (Head)
(PAK467) Projekt “Molecular epidemiology network for promo-
Cure HLH: European initiative to improve knowledge, treatment
Arno Gepp
tion and support of delivery of live vaccines against Theileria
and survival of haemophagocytic syndromes in children
Rolf Müller
parva and Theileria annulata infection in Eastern and Northern
INFLA-CARE: Understanding inflammation-associated tumori-
Africa” (coordination, FZB)
genesis for the rational design of novel anti-cancer therapeutic
Animal Housing
strategies
Dr. Ilka Monath (Head)
Federal Ministry for Education and Research
TB-PAN-NET: Pan-European network for the study and clinical
Exzellenzcluster ‘Entzündung an Grenzflächen’
management of drug resistant tuberculosis
Nationales Genomforschungsnetz 2
Library
POSTICK: Post-graduate training network for capacity building
Christiane Engler
Kai Geschuhn
Verbund ‘Therapie von Infektionskrankheiten unter besonderer
to control ticks and tick-borne diseases Berücksichtigung der Sepsis’
TheSchistoVac: The targeted development of a new generation
Verbund ‘Deutsches Netzwerk für diffus parenchymatöse Lun-
Vaccine for Schistosomiasis
generkrankungen ( GOLD.net)
EUMEDNETvsTB: Building a cooperative strategy between Eu-
Heiko Käßner (Chairman)
PathoGenoMik- Plus Netzwerk / Cluster 3 ‘ Tuberculosis’
rope and Mediterranean Countries for upgrading tuberculosis
Manuel Hein (Vice Chairman)
Förderschwerpunkt ‘Resistenz und Empfänglichkeit gegenüber
research and control
Infektionen’
PiroVac: Improvement of current and development of new vac-
Förderprogramm ‘Ersatzmethoden zum Tierversuch’
cines for theileriosis and babesiosis of small ruminants [coordi-
Verbund ‘Biophotonik’
nation: FZB]
Förderinitiative NanoCare, Forschungsverbund „Prädiktion humantoxikologischer Wirkung synthetischer Carbon Black Nano-
Others
partikel“
Centre for Nanovaccines, Programkomitteen f. Strategiske Vaek-
BMBF Verbundprojekt Med. Infektionsgenomik: Pathogen-Wirts-
stteknologier, Kopenhagen
interaktom und Signalkomplexe in bakteriellen Infektionen
Gates Foundation: Imaging TB
Working Council
81
Administrative and Technical Services
National Networks 2009/2010
Organization chart 2010
Scientific Coordination
Dr. B. Brand
EU-Research Coordination
Dr. K. Lötzer
Department
Molecular Infection Biology
Prof. Dr. U. Schaible
Section Microbiology and
Infection Biology
Prof. Schaible
Cellular
Microbiology
Prof. Schaible
Phagocyte Biology
N. N.
Microbial Ecology
N. N.
Impressum
Herausgeber
Prof. Dr. P. Zabel
Prof. Dr. U. Schaible
Koordination
Dr. B. C. Brand
Bilder
S. 6, 8, 10, 14, 78, 80, 81 Fotolia
Design
STILPUNKT3 Designbüro
Hamburg
Druck
Partner Werbung & Druck
Pinneberg
Medical and
Biochemical
Microbiology
Prof. Brade
Structural
Biochemistry
Prof. Holst
Central Facilities
Board of Directors
Prof. S. Bulfone-Paus
Prof. Dr. U. Schaible
Prof. Dr. P. Zabel
Administration
S. Schrader
Department
Immunology and Cell Biology
Prof. S. Bulfone-Paus
Section Microbial
Inflammation
Research
Prof. Ehlers
Department
Pneumology
Prof. Dr. P. Zabel
Section
Experimental
Pneumology
Prof. Fehrenbach
Mol. Inflammation Immunobiology
Medicine
Prof. Bulfone-Paus
Prof. Ehlers
Vet.-InfectionBiology and
-Immunology
Prof. Ahmed
Infection
Immunology
Dr. Hölscher
Molecular
Immunology
Dr. Lee
Immunoepigenetics
Dr. Singh (temp.)
Microbial
Interface Biology
PD Dr. Reiling
Biochemical
Immunology
Prof. Petersen
Immune
Cell-Analytics
Dr. Ernst
Models of
Inflammation
Prof. Grassl
Biological
Chemistry
Prof. Brandt
Innate Immunity
PD Dr. Heine
Fluorescence
Cytometry
Dr. Scholzen
Section Clinical
Pneumology
Hospital (MCC)
Prof. Zabel
Prof. Zabel
Inflammation and National Reference
Regeneration
Center
Prof. Fehrenbach
Dr. Rüsch-Gerdes
Cellular
Pneumology
Prof. Stamme
Molecular
Mycobacteriology
PD Dr. Niemann
Pathophysiology Clinical and Experiof Inflammation
mental Pahtology
PD Dr. Hauber
Prof. Vollmer
Barrier-Integrity
I. Lautenschläger
Cellular
Allergology
PD Dr. Haas
Clinical and Molecular Allergology
Prof. Jappe
Immunochemistry
Prof. Zähringer
Mucosa
Immunology
PD Dr. Frey
Biophysics
Prof. Gutsmann
Clinical Infectious
Diseases
Prof. Lange
Immunobiophysics
PD Dr. Schromm
Center for
Clinical Studies
Prof. Lange
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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