foot-and-mouth disease virus: linking transmission
TRANSCRIPT
Foot-and-mouth disease virus:
linking transmission pathogenesis
and immunology
Transmission study
2 days later 4 days later 6 days later 48 hrs
inoculates
challenged donors
direct contact recipients
Animals monitored for
14 days post-challenge
In contact
for 24 hrs
FMDV transmission biology
Probang
Nasal swab
Blood
Transmission
Type 1 interferon production (donors)
0
2
4
6
8
10
12
14
0 1 2 3 4 5 6 7 8
Days post infection
U/m
l VN89
VN90
Clinical score VS study day
0
2
4
6
8
10
12
D-2 D-1 D0 D1 D2 D3 D4 D5 D6 D7 D8 D9 D10 D11 D12 D13 D14 D15 D16 D17 D18 D19 D20
Study day
Clin
ica
l s
co
re
VN89
VN90
Type 1 Interferon is found in sera of FMDV infected cattle
at the onset of clinical signs
Data.038
CD172a on CD4+/CD3-
(100,000) stimulated cells
FL2-HC
ount
100
101
102
103
104
0
5
10
15
20
Data.039
MHC II on CD4+/CD3-
(100,000) stimulated cells
FL2-H
Count
100
101
102
103
104
0
5
10
15
20
Data.040
CD11c on CD4+/CD3-
(100,000) stimulated cells
FL2-H
Count
100
101
102
103
104
0
5
10
15
20
Data.042
CD45RB on CD4+/CD3-
(100,000) stimulated cells
FL2-H
Count
100
101
102
103
104
0
5
10
15
20Data.030
FL1-H
FL4-
H
100
101
102
103
104
100
101
102
103
104
13.85%68.18%
16.11%1.85%
CD3
CD 172a MHC II CD 11c CD 45RB
Induction of type-1 interferon by bovine plasmacytoid dendritic cells (pDCs) in response to foot-and-mouth disease virus
Data.005
CD3
CD4
100
101
102
103
104
100
101
102
103
104
Gate 2
4.64%
Gate 2
4.64% 8.88%
79.52% 6.97%
Is the virus entering the cell via the FC receptor? Use antibodies to CD 32 (FC receptor) to block entry
O1K alone O1K+IC
with isotype control
O1K+IC
blocked with anti CD32
Non-structural protein
FMDV capsid protein
0
50
100
150
200
250
300
Medium O1-BFS O1-BFS + immune
IgG
O1-BFS UV O1-BFS UV +
immune IgG
Co
un
ts p
er
min
ute
x 1
03
*
Dendritic cell targeting of FMDV antigen can be improved
Immune response to vaccination rapid induction of antibody variable/ short duration of immunity variable CD4 T cell response
Foot-and-mouth disease virus
Immune response to infection rapid induction of antibody prolonged duration of immunity modest CD4 T cell response
FMDV capsid: T independent and T dependent epitopes
-50
0
50
100
150
200
250
300
350
400
450
500
-1 0 1 2 3 4 5 6 7 9 13 16 21
Control group
Depleted group
Effect of CD4 T cell response on
neutralising antibody response to
vaccination
Persistence of FMDV antigen
4 µm DAPI CD21 FMDV
No expression of non-structural proteins: -Non-replicating -Extracellular
FMDV capsid protein in mouse spleen GC
Structural analysis at the Diamond light source
Structure-based stabilisation of FMDV capsids
Structure-based stabilisation of FMDV capsids
Sum average of aligned images
Capsid stable at 55oC or pH5 for 2 hours
A22_H93C combined – 2 A22_H93C combined – 2
0
200
400
600
800
1000
1200
1400
An
tib
od
y T
itre
Week
Group 1 - Baculovirus WT A capsid
FMD 140
FMD 141
FMD 142
FMD 143
0
200
400
600
800
1000
1200
1400
An
tib
od
y T
itre
Week
Group 2 - Baculovirus mt A capsid
FMD 144
FMD 145
FMD 146
FMD 147
Cattle vaccinated with 12ug of Baculovirus derived FMDV
capsids in commercial oil adjuvant Day 0 and Day 21
0
20000
40000
60000
80000
100000
120000
Media BHK-21 1ug/ml A22 wt Vaccinia
capsid (1μg/ml)
Day 42
WT
mt A
T cell response
Antibody titre
- Stabilise antigen (Persistence of antigen)
- Target antigen - Provide co-stimulation (Danger signals)
Vaccination
Nick Juleff
Julian Seago
Lucy Robinson
Kerry Mclaughlin
Liz Reid
Miriam Windsor
Terry Jackson
Claudine Porta
Dave Stuart
Liz Fry
Ivan Morrison
Mark Woolhouse
Roy Bengis
Francois Maree