infectious disease research in livestock - models, targets ... · infectious disease research in...
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InfectiousInfectious DiseaseDisease ResearchResearch
in in LivestockLivestock
-- Models, Models, TargetsTargets, and , and beyondbeyond --
Christian Menge
Institute of Molecular PathogenesisJena
AnimalAnimal modelsmodels and and relevance/predictivityrelevance/predictivity VeyrierVeyrier du du LacLac, , OctoberOctober 10 10 –– 12, 201112, 2011
Robert Koch (1843 - 1910)
Friedrich Loeffler (1852 - 1915)
Claude Bourgelat(1712 - 1779)
“We have realised the intimacy of the relation which exists between the
human and the animal machines …”
Louis Pasteur(1822 – 1895)
Year Grants reviewed
Grants funding studies using rodents as models
Grants funding studies usingdomestic species as models
Ratio grants w/ rodentvs. domestic species
models
2002 8,842 3,328 79 97.7 : 2.3
2003 10,865 3,898 83 97.9 : 2.1
2004 13,525 3,947 64 98.4 : 1.6
2005 14,877 3,524 52 98.5 : 1.5
2006 16,582 2,262 37 98.4 : 1.6species: mice, rat, hamster, guinea pig, gerbil
species: cow, pig, sheep, horse, chicken, turkey, duck, goat
Ireland et al., J. Anim. Sci., 2008
National Institutes of Health (NIH) grant fundingNational Institutes of Health (NIH) grant fundingfor studies using rodent or domestic species as modelsfor studies using rodent or domestic species as models
2004: First drafts of the bovine and poultry genome
2006: Dog, Bee
2007: Horse
2009: Cattle
2011?? Pig
Current status of genome sequencing Current status of genome sequencing
Courtesy Heiner Niemann, Institute of Farm Animal Genetics
Segment:• smallest distinct part of the lung• one afferent bronchus• individual blood supply
cattle 71sheep 18 man 18
ComparativeComparative anatomyanatomy of of thethe lunglung
Martin’s channels> 120 µm
Lambert’s channels30 µm
Alveolar pores (Kohn)(elevated collateral resistance)
3 - 13 µm
Segmentation Collateral airways
man/dog/ cat/ ferret/ rabbit: existing
horse / sheep: partially existing
cattle / swine: absent !
Courtesy Petra Reinhold
ANTIBODIES 2 IFN-γ elispot kit Endogen 2 IFN-γ polyclonal Endogen 2 IL-1β polyclonal Endogen 2 TNF-α polyclonal Endogen 2 Purified IgG, IgG1, IgG2 Serotec 2 IL-10, IL-1b, IFN-γ, IL-4, IL-6, GM-CSF Serotec 2 IL-12 Serotec 3 IL-4 pairs for ELISA BBSRC 3 IL-6 pairs for ELISA BBSRC 3 IL-10 pairs for ELISA BBSRC 3 IL-12 pairs for ELISA BBSRC 3 IFN-γ pairs for ELISA & kits BBSRC;
Biosource; Biocore/CSL
3 TNFalpha pairs for ELISA BBSRC 3 GM-CSF pairs for ELISA BBSRC 2 CD2, CD4, CD5, CD8, CD11b, CD21, CD26,
CD63, CD172a, CD62L, CD205, PL-2p85β, cyclin A, IgA, IgG, IgG1, IgG2, IgM
Serotec
2 CD1a, CD2, CD3, CD4, CD5, CD6, CD8, CD11a-c, CD14, CD18, CD21, CD25, CD41/CD61, CD44, CD45, CD45R, CD45RO, CD58, CD62L, CD71, WC1-15
(prepared by Chris Howard)
2,3 IgG1, IgG2, IgA, IgM, Ig(H+L), GM-CSF, IL-1β, MHC class I, MHC class II, TCR-δ, WC1, CD1, CD2, CD3, CD4,CD5,CD6,CD8α,CD11a/CD18, CD11b, CD11c, CD14, CD21, CD25, CD26, CD29, CD41/CD61, CD44, CD45, CD45R, CD45RO, CD49d, CD62L, CD172a
VMRD, Pullman, WA
Available Reagents and Level of AccessibilityAccessibility Level:
1=Totally available-e.g. ATTC, or distributed without strings2= Commercially available, thus accessible at cost3= available through collaborative agreement only
(either purified or recombinant protein or if antibody is designated with “?”)
Reagent: Source:
PROTEIN2 IL-4 Endogen2 IL-6 Endogen2 IFN-tau Endogen2 IFN-? Endogen2 IGF-1 Endogen3 IL-2 BBSRC 3 IL-4 BBSRC3 IL-5 BBSRC3 IL-6 BBSRC3 IL-10 BBSRC3 IL-12 BBSRC3 IFN-g BBSRC3 GM-CSF BBSRC2 IFN-tau Cell sciences
US BiologicalsPBL Biomedical Labs
US VETERINARY IMMUNE REAGENT NETWORK (coordinator C. Baldwin) http://www.umass.edu/vetimm/
Bridger et al., Vet. Immunol. Immunopathol, 2010
EDL933- O157:H7- stx1 (+)- stx2 (+)- eae (+)
E2348/69- O127:H6- eae (+)- espB (+)- ehx (+)
C600- apathogeniclab strain
field isolate- O101:K28- cnf1/2 (-)- esta (-)- stx1 (-)- stx2 (-)- eae (-)- fan (-)- fim41a (-)- f17A (-)- papC (-)- iucD (-)
914/06-1- O101:K28- esta (+)- fan (+)- fim41a (+)
STEC (EHEC) EPEC K12 P391
Primary bovine colonocytes: bacterial adhesion
Colonic crypt culture (n=2)
Exposure tobacterial strains(duplicate cultures)
Medium change
FAS-Test
Giemsa staining4 days 3 h 3 h
ETEC
ETEC
Lotte – from the left Lotte – from the right
Experimental setup for examination of rumen metabolismExperimental setup for examination of rumen metabolism
Courtesy Sven Dänicke, Institute of Animal Nutrition
• Infection:– C. psittaci DC15 or C. abortus S26/3
– on embryonic developmental day 10
– 5x104 ifu/egg
– vitality and sampling: 1 dpi - 8 dpi
• Samples:
– Chorionallantoic membrane (CAM)
– internal organs: liver, spleen, bursa of Fabricius, heart, stomach, intestine, brain
EmbryonatedEmbryonated egg egg modelmodel of of chlamydialchlamydial infectioninfection
Courtesy Maria Braukmann / Angela Berndt
ClinicalClinical haematologyhaematology and and biochemistrybiochemistry
e.g. blood gas analysis
Courtesy Petra Reinhold
Vt
IC VC
RV RV
IRV
FRC
TLC
ERV
Animals with body masses 50-100 kg (e.g. calves, pigs, sheep)
Physiological range of key parameters(volumes, flow, resistance)
comparable to humans
Identical techniques applied:
non-invasiv
applicable to animals and man under conditions of spontaneus breathing
LungLung functionfunction assessmentassessment
Courtesy Petra Reinhold
dpa / FLI
Impulse Impulse oscillometryoscillometry (IOS)(IOS)
H.-J. Smith
FLI
FLI
Courtesy Petra Reinhold
0
2
4
6
8
10
-2 4 10 16 22 28 34 40 46 52 58
L1-controlL2-infect.
Median -/+ min/maxCD25 on CD4+/CD45RO+ cells
0
2
4
6
8
10
-2 4 10 16 22 28 34 40 46 52 58
L1-controlL2-infect.
CD25 on CD2-/γδT+ cells
Weeks post inoculation
Rea
ctio
ntit
er[x
-fold
vers
usM
. phl
ei]
Antigen-induced CD25 expression by bovine lymphocyte subsetsex vivo
Rea
ctio
ntit
er[x
-fold
vers
usM
. phl
ei]
Antigen-induced CD25 expression by bovine lymphocyte subsetsex vivo
0
2
4
6
8
10
12
-2 4 10 16 22 28 34 40 46 52
controlcalvesMAP-inf.calves
0
2
4
6
8
10
-2 4 10 16 22 28 34 40 46 52 58
L1-controlL2-infect.
Median -/+ min/maxCD25 on CD4+/CD45RO+ cells
Weeks post inoculation
18 hpi, distal jejunum
Otto et al., J. Virol., 2011
Pathohistologyand distribution of Jena virus antigen
Detection ofJena virus particles
in fecal samples
Experimental Experimental NorovirusNorovirus infection of newborn calvesinfection of newborn calves
Diarrhoea(14 – 16 hpi until 69 – 83 hpi)
Infection: oro-nasal (secretions, aerogenic)
Neuroinvasion of Neuroinvasion of PseudorabiesPseudorabies virusvirus ((PrVPrV))
Courtesy Thomas C. Mettenleiter, Institute of Molecular Biology
Ekstrand et al., Trends Mol. Med., 2008
AlphaAlpha--herpesvirusesherpesviruses::molecularmolecular pathfinderspathfinders in in nervousnervous systemsystem circuitscircuits
Livestock disease modelsLivestock disease modelsInfection / Disease Host(s) Human disease Examples of aspects studied
Influenza Swine, horses Influenza Pathogenesis of clinical illnessRotavirus Cattle, swine viral diarrhoea Pathogenesis of clinical illnessNorovirus Cattle Norovirus infection Pathogenesis and vaccine developmentRespiratory syncytial virus (BRSV) Cattle RSV infection Vaccine developmentCaprine arthritis encephalitis virus Goats, sheep HIV Genetic susceptibility andand Visna/maedi virus lentivirus–host adaptationBovine leukemia virus Cattle, sheep human T-cell leukaemia Oncogenesis
virus (HTLV) infectionBovine papilloma viruses Cattle Human papilloma viruses Latency mechanismsAujeszky´s disease Swine Human herpes virus infections Latency mechanismsMarek´s disease virus Chickens virus-induced lymphoma Oncogenesis
Hepatitis E Swine Hepatitis E Pathogenesis and vaccine development
Scrapie Sheep transmissible spongiform Neurogenerative disordersencephalopathy (TSE)
Bovine spongiform encephalopathy Cattle variant Creutzfeld-Jakob disease Neurogenerative disorders
Mycoplasma bovis / hyopneumoniae Cattle / swine Mycoplasma pneumoniae Pathogenesis of clinical illness
Chlamydia psittaci Cattle Psittacosis Mechanisms of acute disease and persistence
Tuberculosis (M. bovis) Cattle, deer Tuberculosis (MTC) Latency mechanismsJohne´s Disease Cattle, sheep, Inflammatory bowel diseases Genesis of chronic immunopathological
goats, deer (e.g. Crohn´s disease) disorders (chronic inflammation)MRSA Swine pulmonary MRSA infections Therapeutic treatment
Cryptosporidium parvum Cattle, swine Cryptosporidium parvum Therapeutic treatment testing andclinical responses to diverse strains
Asthma Sheep Asthma Genesis of chronic immunopathological(Ascaris antigen-induced) disorders
Esse
ntia
lN
on-e
ssen
tial
gB
gC
gD
gH/L
gK
gE/I
gG
gM/N
Antibodies
+
+
+++
+++
+
+
+
++
+
+++
+
+
+
+
-
Courtesy Thomas C. Mettenleiter, Institute of Molecular Biology
DIVA DIVA ((ddifferentiating ifferentiating iinfected from nfected from vvaccinated accinated aanimals)nimals) vaccinesvaccines
Pseudorabies virus
SecondSecond--generation licensed veterinary viral vaccinesgeneration licensed veterinary viral vaccines
Meeusen et al., Clin. Microbiol. Rev., 2003
ZoonosesZoonoses
Bauerfeind mod. from Woolhouse & Gowtage-Sequeria, 2005
Total number of known human pathogens: 1.407
Zoonotic pathogens
Pathogens specific for humans
„Emerging /“Re-emerging Diseases“:
177 pathogens816 (58 %)
591 (42 %)of thesezoonotic pathogens: 130
of these pathogensspecific for humans: 47
Lanzas et al., Nature Rev. Microbiol., 2010
Farm animal populationsFarm animal populationsas models for infectious diseases of humansas models for infectious diseases of humans
27 / 27 calves (min. tested until 9th week of life)
stx1 stx2
p. n. 24 h 1 2 3 4 5 6 7 8 9 10 1211 17/18 21/2213/14 15/16 19/20 23/24 dama. p.
damp. p.
Week of life
123456789
101112131415161718192021222324252627
Calf no.
Fröhlich et al., 2009, Appl. Environ. Microbiol.
STEC (EHEC) shedding in calves STEC (EHEC) shedding in calves ((stxstx detection in detection in faecalfaecal samples)samples)
eine wirksame Bekämpfung von
0
500
1000
1500
2000
2500
3000
34 36 38 40 42 44 46 48 50 52 2 4 6 8 10 12 14 16 18 20 22 24 26 28 30 32 34 36 38 40 42 44 46 48 50 52 2 4 6 8 10 12 14 16 18 20 22 24 26 28 30 32 34 36 38 40 42 44 46 48 50 52 2 4 6 8 10 12 14 16 18 20 22 24 26 28 30 32 34 36 38 40 42 44 46 48 50 52
Calendar week
Num
ber o
f cas
es/o
utbr
eaks
Goats
Sheep
Cattle
Source: TSN, 2010-03-16
20072006 2008 2009
Start vaccination21 calendar week 2008
Vaccination campaign
2008
Vaccination campaign
2009
BluetongueBluetongue DiseaseDisease (BTV) in Germany(BTV) in Germany
Courtesy Thomas C. Mettenleiter, Institute of Molecular Biology
2007: 20.813 cases/outbreaks
Wolfe et al., Nature Reviews, 2007
Origins of major human infectious diseasesOrigins of major human infectious diseases
chemokineinduction
STEC
CD8αβ+
CD21
CD21+
CD77dim
CD8αβ+
CD77dim
CD14+
CD8αβ+ CD8αβ+
IL-4
chemokineinductionIL-10
NOchemokinemodulationX
(mucosal) macrophage
X
XCD80/CD86
IL-4IL-6
TNF-α
phagocytosis, oxidative burst
Stx
ShigatoxinsShigatoxins modulate the modulate the immune response in cattleimmune response in cattle
Models• Long-lived
Chronic diseases• Diagnostic approaches
comparable to human medicineTranslational medicine
• Parallel assessment of a plethora of parameters (individually and over time)
Systems biology• Traditional models
Multitude of archiveddata available togenerate hypotheses
• Biologically more relevant,less animals needed
economic,more ethical
Infectious Disease Research in LivestockInfectious Disease Research in Livestock
Targets• Natural host species for the
causative agents of animal epidemics and zoonoses
Prototypic strains“out of the field”Specific host-pathogeninteractionsVaccine developmentEpidemiologicalmodeling
• Diagnostic approaches identical to routine diagnostics
Eradication programs
• Biologically more relevant,less animals needed
economic,more ethical
beyond• Defined and traceable
populationsModels for transmissionof diseases(including wildlife-derived and vector-borne)
• Natural (intermediate) reservoir species of zoonotic pathogens
Evolution of novelpathogens
• Biologically more relevant,less animals needed
economic,more ethical
ThanksThanks to to ……Thomas Mettenleiter
Institute of Molecular BiologyMartin Groschup
Institute of Novel and Emerging Infectious DiseasesTim Harder, Martin Beer
Institute of Diagnostic VirologyHeinrich Niemann
Institute of Farm Animal GeneticsSven Dänicke
Institute of Animal NutritionPetra Reinhold, Elisabeth M. Liebler-Tenorio, Carola Ostermann, Maria Braukmann, Angela Berndt
Institute of Molecular Pathogenesis
Jürgen A. RichtCollege of Veterinary Medicine
Helge KarchInstitute for Hygiene
Rolf Bauerfeind, Georg Baljer, Philip S. BridgerInstitut for Hygiene and Infectious Diseases of Animals
Klaus Doll, Torsten SeegerClinics for Ruminants and Swine