Cyril gay nfid vaccine research conference, fmd vaccines, april 2013
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Transcript of Cyril gay nfid vaccine research conference, fmd vaccines, april 2013
16TH ANNUAL CONFERENCE ON VACCINE RESEARCH
16TH ANNUAL CONFERENCE ON VACCINE RESEARCH
DEVELOPMENT OF VACCINES TOWARD THE GLOBAL CONTROL AND ERADICATION OFFOOT-AND-MOUTH DISEASE (FMD)
Cyril G. Gay, DVM, PhD owns stock from Pfizer Inc.
Development of vaccines toward the
global control and eradication of Foot-
and-Mouth Disease (FMD)Cyril Gerard Gay, DVM, PhDSenior National Program LeaderAnimal Production and Protection
Agricultural Research [email protected]
Presentation Outline
1. Importance of Animal Agriculture2. Disease Threats3. Cost of FMD 4. FMD Eradication5. FMD Virology and Pathogenicity6. FMD vaccines7. Conclusions
33
Importance of Animal Importance of Animal AgricultureAgriculture
• FAO estimates that livestock contribute 40% of the global value of agricultural output and support the livelihoods and food security of almost 1 billion people
44
2121stst Century Challenges Century Challenges
• World population is projected to reach 9 billion
• Global food production will need to double in order to meet these food demands.
• 73% increase in consumption of animal protein
55
58% increase in consumption of dairy products
Presentation Outline
1. Importance of Animal Agriculture2. Disease Threats3. Cost of FMD 4. FMD Eradication5. FMD Virology and Pathogenicity6. FMD vaccines7. Conclusions
66
List of 17 Most Damaging List of 17 Most Damaging Animal Disease ThreatsAnimal Disease Threats
1. Highly Pathogenic AI (F) 2. Foot-and-Mouth Disease 3. Rift Valley fever (F) 4. Exotic Newcastle Disease 5. Nipah and Hendra virus
(F) 6. Classical swine fever 7. African swine fever 8. Bovine spongiform
encephalopathy (F) 9. Rinderpest (E) 10. Japanese encephalitis (F)
11. African horse sickness12. Venezuelan equine (F)
encephalitis 13. Contagious bovine
pleuropneumonia 14. Ehrlichia ruminantium
(Heartwater)15. Eastern equine
encephalitis (F)16. Coxiella burnetii (F)17. Akabane virus
F: Potentially fatal to humansYellow text: FBI pathogens of ConcernE: Eradicated
H5N1 Avian Influenza VirusSource: PHIL CDC
Emerging Diseases(and re-emerging diseases)
Human Animal• HIV/AIDS• Ebola*• Hantaan• Legionaire’s disease• BSE*• SARS*• Dengue• West Nile*• Nipah virus*• Rift Valley Fever*• Chikungunya virus• H5N1, H3N2v, H7N9*• pandemic H1N1
• BSE*• CWD• West Nile*• Foot-and-Mouth Disease• Classical Swine Fever• Blue Ear Pig Disease• Rift Valley Fever*• Avian Influenza H5N1/H7N9*• Nipah and Hendra*• Bluetongue• African Swine Fever• African Horse Sickness• pandemic H1N1**
* Zoonoses **Reverse Zoonosis
Presentation Outline
1. Importance of Animal Agriculture 2. Disease Threats3. Cost of FMD 4. FMD Eradication5. FMD Virology and Pathogenicity6. FMD vaccines7. Conclusions
99
HistoryHistory
• Over 100 years of research in FMDV
• 1924-British Minister of Agriculture appointed a committee “to initiate, direct and conduct investigations into FMD… discovering means whereby the invasion of the new disease may be rendered less harmful to agriculture…”
(from B.W. Mahy, 2005)
• Successful eradication in Europe
• US free since 1929
In 1898, Freidrich Loeffler and Paul Frosch showed that a virus was responsible for foot-and-mouth disease in cattle
1010
The Cost of FMDThe Cost of FMD
• Total loss of 2001 outbreak in the United Kingdom was estimated to be between $12.3 and $15 billion (US$)
• 36% was lost tourism , Slaughter of 6.5 million livestock• $4.2 billion paid by government in compensation to the
agriculture and food industry• Social effects, Human cost (suicides)• For some countries mass slaughter is NO LONGER an
option!! (e.g. S. Korea, 2011)
The Cost of FMDThe Cost of FMD• On the global scale FMD causes
damage and hampers development : Cost USD 5 billion per year
• Outbreaks in FMD-free countries worldwide costs USD 1 billion/year
• The world is a global village; risks for FMD-free countries will only increase
• Fighting the disease at source should be part of the prevention strategy of FMD-free countries(Rushton, 2012)
Presentation Outline
1. Importance of Animal Agriculture2. Disease Threats3. Cost of FMD 4. FMD Eradication5. FMD Virology and Pathogenicity6. FMD vaccines7. Conclusions
1313
Pool 7
O, A
Pool 5 O, A, SAT 1, 2
Pool 6 SAT 1, 2, 3
Pool 4 A, O, SAT 1, 2, 3
Pool 2 O, A, Asia 1
Pool 1 O, A, Asia 1
Pool 3 O, A, Asia 1
Endemic
Free with vaccination
Intermediate, sporadic
Countries with multiple zones:FMD-free, free with vaccination or not free
Free. Virus present in game parks
Free Pool positions are approximate and colours indicate that there are three principal pools, two of which can be subdivided into overlapping areas
Status of FMD showing approximate distribution of regional virus endemic pools
Institute for Animal Health-WRLFMD®
Website: http://www.fao.org/ag/againfo/commissions/eufmd/commissions/eufmd-home/progressive-control-pathway-pcp/en/
FAO/OIE Progressive Control Pathway
for the control of FMD
Within a 15-year period:
1) countries that are currently in PCP Stages 0 and 1 will have progressed at least two stages along the PCP
2) countries in PCP Stages 2 or 3 should also move up two stages, but the final objective will depend on a country’s decision based on cost-effectiveness studies
3) countries or zones that already have an OIE-recognized FMD-free status maintain this status or further improve it (i.e. go from FMD-free with vaccination to FMD-free without vaccination)
Objectives of FMD Control Strategy
- Cost of national FMD programs for 79 initial 0-2 Stage countries: 68 M- Vaccination cost for 45 initial 1-3 Stage countries (excluding India and China): 694 M- Regional level (ref. lab and epidemiology support and networks) 47 M- Global level (coordination, evaluation) 11 M
Financial implications (first 5 years)
(in USD as calculated by the World Bank)
Presentation Outline
1. Importance of Animal Agriculture 2. Disease Threats3. Cost of FMD 4. FMD Eradication5. FMD Virology and Pathogenicity6. FMD vaccines7. Conclusions
1818
Features of FMDV
25 nm
• Family Picornaviridae, genus aphtovirus
• Positive sense RNA Approximately 8.2 kb
• Seven serotypes: A, O, C, Asia, Sat1, Sat2, Sat3
P1 P2
poly(A)
3'UTR
3B
IRESS
5'UTR
PKs
crepoly(C)
3B1231A
Lpro 1B 1C 1D
2A
2B 2C 3A 3Cpro 3Dpol
P3
**
Protease Cleavage Sites
Lpro unknown
2A3Cpro
2 in-frame AUGs **
FMDV genome
VP0(1AB)
3AB123
structural proteins nonstructural proteins
partial partial cleavagecleavageproductsproducts
P1/2A P2BC P3
3B123CD1ABC 3A
L
VP1
VP3
2C2B
3A 3B13B23B3 3C
3CD
3D
FMDV-Key Information
• Systemic disease of domestic and wild cloven-hooved animals
• Acute disease characterized by fever, lameness, and vesicular lesions on the feet, tongue, snout, and teats
• FMD is considered to be one of the most contagious infectious disease known
2121
FMDV-Key Information• Multiple subtypes reflect significant genetic
and antigenic variability • Some strains of the virus and some host
species show minimal or no signs of disease• The emergence of new variants of FMDV is
common• Fifty percent of infected cattle become carriers• The pathogenesis of FMDV, including
mechanisms of viral transmission and the carrier state, are not fully understood
• The early detection of FMDV is paramount to stop the spread of the virus and disease and reduce economic impact
2222
Aerosol Inoculation ModelAerosol Inoculation Model
The nebulizer consists of a commercially available aerosol delivery system that will produce an average of particles of 5 microns (Hess et al., 1996) and a large equine mask. Entire respiratory tract is exposed to virus.
Pacheco et al, 2008
Dose: FMDV 107 TCID50
2323
Summary of FMD Early Pathogenesis in Cattle
V V V V
V VV VV
V
V V
SystemicCirculation
Aerosol ExposureT = 0.1 HPA T = 3 – 6 HPAT = 12 HPAT = 24 HPAT = 48 HPA
24
FMD-BovineMouth
25
FMD-BovineHoof
26
Presentation Outline
1. Importance of Animal Agriculture2. Disease Threats3. Cost of FMD 4. FMD Eradication5. FMD Virology and Pathogenicity6. FMD vaccines7. Conclusions
2727
FMDV- Commercial Vaccines
• Conventional inactivated vaccines have been successfully used in disease eradication programs in endemic areas of Africa, South America, and Europe
• Requires adaptation of wild type virus to cell culture• Virulent virus grown on BHK cells• Production of large volume requires BSL-3 facilities• Virus yield sometimes low for hard-to-adapt viruses• Inactivated with binary ethyleneimine (BEI)• Non-structural proteins removed• Non-formulated bulk fluids are stored frozen for stockpiling• Adjuvanted with alum or oil emulsion• Vaccines provide fail to induce long lasting immunity
2828
T.R. Doel / Virus Research 91 (2003) 81/99 86
Current Vaccines
29
Inactivated Vaccine
FMDV- Vaccine Information
• FMDV Serotype O is less immunogenic• FMDV Serotype O is more prevalent in
South America • Vaccines for FMDV Serotype O need a
higher payload than Serotypes A, C, Asia, or SAT
• FMDV Serotypes SAT1, SAT2, SAT3 antigens are less stable
• FMDV Serotypes A and SAT 2 are more hypervariable than other serotypes
3030
Risk of Vaccine Production Risk of Vaccine Production with Virulent FMDV with Virulent FMDV
On Friday August 3, 2007 FMD was detected in a farm in Southern England located within 6 miles of the Pirbright Laboratory site Outbreak resulted in trade barriers and billions $$ loses
3131
Concerns with FMD Vaccines in Disease-Free Countries
• Require adaptation and growth of large volumes of wild type virus in cells
• Escape of virus from manufacturing facilities• Require banking of multiple antigen concentrates • Some antigens lack stability (low potency/short shelf
life)• Onset of protection 7-14 days• Short duration of immunity <6 months • Difficult to differentiate vaccinated from infected
animals (DIVA) due to presence of NS proteins• Vaccinated and exposed animals become carriers
3232
Characteristics of the “Ideal” FMD Vaccine
• Effective, rapid and long-lasting protection with one inoculation
• Prevents viral transmission• Allow differentiation of infected from vaccinated
animals (DIVA)• Produced without the need for virulent FMDV• Prevent development of carrier state• Protection against multiple serotypes• Stable antigen – long shelf life• Reasonable cost to enable eradication programs
Adenovirus-Vectored FMD Vaccine
Expressing Empty Viral Capsids
• Contains all protective epitopes present on current inactivated virus vaccine but lacks infectious viral nucleic acid and non-structural protein (NSP)
• Allows to “clearly” distinguish vaccinated from infected animals using 3D and other NSP diagnostic tests
• Can be safely produced in the United States
“Left-out” proteins can be used for DIVA tests
Processed products display
epitopes resembling
intact capsid.
3D
FMDV Empty Capsid Vaccine
P1 2A 2B’ 3B’ 3C
VP0 VP3 VP1
3C
L P1 2A 2B 2C 3A 3B 3C
Remove regions unnecessary for
capsid formation.
3D
DHS TAD ProgramLicensed by CVB-APHIS
2 Negative markers: DIVA tests
3Dpol ELISA
3B ELISA
3B233’NTR
IRESS
1A
1B 1C 1D
2A
2B 2C 3A 3C 3D A
poly(C)
NON-STRUCTURALSTRUCTURAL
L
5’NTR
RE1 RE2
Easy swap of capsid sequences
Vaccine seed antigens
Deletion of Leader protein (543 bp)
Attenuating factor
FMD-LL3B3D: A Safe Platform For FMD Vaccine Production With Built-In DIVA Markers Key Features
36
FMD Vaccine Product Profiles : FMD Vaccine Product Profiles : Current Inactivated versus Current Inactivated versus Inactivated FMD-LL3B3DInactivated FMD-LL3B3D
NoNoProvides cross-serotype protection
NoCompatible with “vaccinate to live” strategy
NoNoReadily deployable (ready to use)
NoNoLong duration of immunity
PossibleNoDomestic production (USA)
+/-Marked vaccine (DIVA capable)
Early onset of immunity (7 DPV)
Prevents viral transmission
MOLECULAR INACTIVATED
CURRENT INACTIVATED
PRODUCT PROFILE
Possible
Proteinase Proteinase domain/De-domain/De-ubiquitinaseubiquitinase
Topology, Topology, DNA binding, DNA binding, transcription transcription regulationregulation
Topology, Topology, interaction with interaction with phosphoproteins, phosphoproteins, signaling signaling interferenceinterference
Interaction Interaction with eIF4Gwith eIF4G
SAP FHA-likeProteinase
1 29 75 112 167 183 201
N C
Lab Lb
Bioinformatics analysis suggests the presence of multiple domains
(by Dr. James Zhu)
Disruption of the L protein SAP domain results in attenuation in vitro
SAP domains are conserved protein domains present in eukaryotic nuclear proteins involved in chromosomal organization and repression of transcription.
Double mutation of FMDV L protein SAP domain results in:
Altered L protein sub-cellular distribution: L SAP mutant localizes only to the cytoplasm of infected cells by 6 hpi while L wild type is in the cytoplasm and nucleus.
L SAP mutant is unable to cause degradation of NF-κB inducing higher levels of IFN, inflammatory cytokines and chemokines in comparison to WT.
(de los Santos et al., 2009)
00 11 22 33 44 55 66 77dpidpi
Virus intradermal inoculation in Virus intradermal inoculation in right rear foot-padright rear foot-pad
TemperatureSerumNasal Swabs
Temperature Plasma/Serum Clinical signs Nasal Swabs
1414 2121
x3x3 x3x3 x3x3xx33
x3x3
Group #1:Group #1: Group #2:Group #2: Group #3:Group #3: Group #4:Group #4: Group #5:Group #5:
FMDV A12-WT1x105 pfu/pig
FMDV A12-WT1x106 pfu/pig
FMDV A12-SAP1x105 pfu/pig
FMDV A12-SAP1x106 pfu/pig
FMDV A12-SAP1x107 pfu/pig
Serum
Does disruption of the L protein SAP Does disruption of the L protein SAP domain results in attenuation domain results in attenuation in vivo?in vivo?
Clinical Score
pfu
/ml
0
2
4
6
8
10
12
14
16
18
0dpc 1dpi 2dpi 3dpi 4dpi 5dpi 6dpi 7dpi
10e5 A12-SAP
10e5 A12-WT
10e6 A12-WT
10e6 A12-SAP
10e7 A12-SAP
Viremia
Nasal swabs
0dpi 1dpi 2dpi 3dpi 4dpi 5dpi 6dpi1.0E+00
1.0E+01
1.0E+02
1.0E+03p
fu/m
l
0.0E+00
4.0E+02
8.0E+02
1.2E+03
1.6E+03
0dpi 1dpi 2dpi 3dpi 4dpi 5dpi 6dpi 7dpi
A12-SAP mutant is attenuated A12-SAP mutant is attenuated in vivoin vivoS
co
re (
ma
x.
17
)
Animals inoculated with A12-SAP are Animals inoculated with A12-SAP are completely protected when challenged completely protected when challenged with WT FMDV at 21 dayswith WT FMDV at 21 days
Group Animal Challenge virus at 21dpi
Dose Viremia (dpc, day of onset, duration)
PFU in nasal swabs (dpc, day of onset,
duration)
Neutralizing antibodies PRN70
7dpc
A12-SAP 1x105pfu/pig
90 A12-WT 1x105 Neg. Neg. >3.1
91 Neg. Neg. >3.1
92 Neg. Neg. >3.1
A12-SAP 1x106pfu/pig
93 A12-WT 1x105 Neg. Neg. >3.1
94 Neg. Neg. >3.1
95 Neg. Neg. >3.1
A12-SAP1x107pfu/pig
96 A12-WT 1x105 Neg. Neg. 3.0
97 Neg. Neg. 3.1
98 Neg. Neg. >3.1
x3x3
x3x3
x3x3
FMDV A12-SAP1x106 pfu/pig
FMDV A12-SAP1x106 pfu/pig
FMDV A12-SAP1x106 pfu/pig
CONTROLPBS
x3x3
x3x3
FMDV A12-SAP1x106 pfu/pig
VACCINATION DOSE
TIME OF CHALLENGE
CHALLENGE DOSE
14dpv14dpv
7dpv7dpv
4dpv4dpv
2dpv2dpv
14dpi14dpi
FMDV A12-WT5x105 pfu/pig
FMDV A12-WT5x105 pfu/pig
FMDV A12-WT5x105 pfu/pig
FMDV A12-WT5x105 pfu/pig
FMDV A12-WT5x105 pfu/pig
SAP mutant vaccination experiment with SAP mutant vaccination experiment with early challenge in swineearly challenge in swine
Inoculation with FMDV A12-SAP confers Inoculation with FMDV A12-SAP confers protection as early as 2 days post protection as early as 2 days post vaccinationvaccination
Vir
us
in s
eru
m o
r n
asal
sw
abs
(p
fu/m
l)
Clin
ical
Sco
re
1.0E+00
1.0E+01
1.0E+02
1.0E+03
1.0E+04
1.0E+05
1.0E+06
0dpc 1dpc 2dpc 3dpc 4dpc 5dpc 6dpc 7dpc0
2
4
6
8
10
12
14
16
18
14 dpv
4 dpv
7 dpv
2 dpv
control
viremia NScs
SummarySummary
A12-SAP mutant is avirulent in swine but induces a strong neutralizing antibody response
In vivo attenuation correlates with increased levels of pro-inflammatory cytokines whose transcription
depends on NF-κB
Vaccination of swine with A12-SAP results in complete protection against homologous challenge
as early as 2 days post-inoculation, when no adaptive immune response is detectable
Conclusions
1. Importance of Animal Agriculture2. Disease Threats3. Cost of FMD 4. FMD Eradication5. FMD Virology and Pathogenicity6. FMD vaccines
4646
www.ars.usda.gov/research/programs/programs.htm?NP_CODE=103Publications
www.ars.usda.gov/GFRA/
Global Foot-and-Mouth Disease Research Alliance
4747
AcknowledgementAcknowledgement
• Luis Rodriguez• Marvin Grubman• Jonathan Arzt• Juan Pacheco• Elizabeth Rieder• James Zhu• Teresa de los Santos• Bill Golde
4848
Thank you!
Thank you !!!