Download 30. Evaluation of cross-protection between O1 Manisa and O1 Campos in cattle vaccinated with O1 Manisa vaccines

Appendix 30
Objectives
EVALUATION OF CROSS-PROTECTION BETWEEN
O1 MANISA AND O1 CAMPOS IN CATTLE
VACCINATED WITH O1 MANISA VACCINES
V.A.Srinivasan1*, S.B.Nagendrakumar1, M.Madhanmohan1, S.Yuvaraj1,
S.Parida2, Antenello di Nordo2, D.J.Paton2,
1 Indian Immunologicals Limited, Rakshapuram, Gachibowli Post, Hyderabad 500
019, India
2 Pirbright Laboratory, Institute for Animal Health, Ash Road, Woking, Surrey
GU24 0NF, UK
[email protected]
• Serology is used to predict vaccine induced protection against
challenge with a heterologous strain of the same serotype of
foot-and-mouth disease virus (FMDV)
• To evaluate the accuracy of such predictions the protection
afforded to cattle vaccinated with different payloads of the O1
Manisa strain of FMDV against challenge with either a
homologous (O1 Manisa) or a heterologous strain (O1
Campos) was compared (FP6 grant SSPE-CT-2003-503603).
• Serology by virus neutralization test (VNT) using O1 Manisa
antiserum predicted an acceptable protection (r1 = 0.60)
against such a challenge
Experimental Design
• Oil adjuvant monovalent vaccines with O1 Manisa (2ml/dose)
were prepared with the following payloads
–
–
–
–
0.94 µg
3.75 µg
15 µg
60 µg
• Preparation of cattle challenge viruses
– O1 Manisa
– O1 Campos
• Cattle challenge studies using modified potency test
– Homologous potency test using blends 15, 3.75 and 0.94µg
– Heterologous potency test using blends 60, 15, 3.75 µg
Results of percentage protection against homologous challenge with O1 Manisa on
21 dpv in cattle calves vaccinated with different payloads of O1 Manisa vaccine
Experiment 1
Experiment 2
Groups
Payload
(μg)
Group 2
15
8
8
100
5
5
100
Group 4
3.75
8
8
100
5
5
100
Group 6
0.94
8
7
88
5
1
20
Group 8
UV
Control
8
0
0
2
0
0
Number
Number Percentage
Number
Number Percentage
Challenged Protected Protection Challenged Protected Protection
“New tools and challenges for progressive control”
Open Session of the EuFMD Research Group, Vienna (Austria) 29 September ‐ 1 October 2010
Appendix 30
Results of percentage protection against heterologous challenge with O1 Campos on
21 dpv in cattle calves vaccinated with different payloads of O1 Manisa vaccine
Experiment 1
Experiment 2
Number
Number Percentage
Challenged Protected Protection
Number
Number Percentage
Challenged Protected Protection
Groups
Payload
(μg)
Group 1
60
8
6
75
5
5
100
Group 3
15
8
4
50
5
3
60
Group 5
3.75
7
2
29
5
0
0
Group 7
UV
Control
8
0
0
2
0
0
Serum antibody response towards O1 Manisa vaccine in animals challenged with
(a) O1 Manisa and (b) O1 Campos
Experiment 1
Mean rectal temperature in vaccinated and control animals
Homologous challenge
Homologous challenge
Experiment 2
Mean of copy numbers of FMDV detected at 5, 10, 15, 21 and 35 days post-challenge
by quantitative RT-PCR with different Ag payloads used for homologous
(a) and heterologous (b) challenge. Error bars represent 95% CI.
“New tools and challenges for progressive control”
Open Session of the EuFMD Research Group, Vienna (Austria) 29 September ‐ 1 October 2010
Appendix 30
Scatter plot of O1 Manisa serological responses at 21 days post-vaccination and proportion protected against
homologous and heterologous challenge with fitted lines for the RTO models and corresponding 95% CIs.
Data from both experiment 1 and 2 were included to plot the graphs.
Scatter plot of O1 Campos a serological responses at 21 days post-vaccination and proportion protected against
homologous and heterologous challenge with fitted lines for the RTO models and corresponding 95% CIs.
Data from both experiment 1 and 2 were included to plot the graphs.
PA50 for O1 Manisa with O1 Manisa VNT results: 0.963
PA50 for O1 Campos with O1 Manisa VNT results: 1.741
PA50 for O1 Manisa with O1 Campos VNT results: 0.879
PA50 for O1 Campos with O1 Compos VNT results: 1.771
Probit probability analysis of serum antibody titers for PA50 values
PA50 for O1 Manisa: 0.879
Probit probability analysis of antigen payload for PD50 values
PD50 for O1 Manisa: 0.92 mcg/dose
PA50 for O1 Campos: 1.739
PD50 for O1 Campos: 32.2 mcg/dose
B cell epitopes in 1D region
Sequence identities of the O1 Manisa viruses used in vaccine
production, cattle challenge test and neutralisation test
OMANISA69 TTSAGESADPVTATVENYGGETQVQRRQHTDVSFILDRFVKVTPKDQINVLDLMQTPAHT
O1CAMPOS55 TTSAGESADPVTTTVENYGGETQIQRRQHTDVSFIMDRFVKVTPQNQINILDLMQIPSHT
OMANISA69 LVGALLRTATYYFADLEVAVKHEGNLTWVPNGAPEAALDNTTNPTAYHKAPLTRLALPYT
O1CAMPOS55 LVGALLRASTYYFSDLEIAVKHEGDLTWVPNGAPEKGLDNTTNPTAYHKAPLTRLALPYT
9
T
P
P
P
P
P
P
VP4
40
D
D
D
E
E
E
E
OMANISA69
O1CAMPOS55
KRAETYCPRPLLAIHPDQARHKQKIVAPVKQLL
KRAETYCPRPLLAIHPTEARHKQKIVAPVKQTL
BHK7
BHKCZ
Cattle
Passage 1
Cattle
Passage 2
Affected
cattle - 3930
Feet
Epithelium
Amino acid
Position
OMANISA69 APHRVLATVYNGNCKYGDGTVANVRGDLQVLAQKAARALPTSFNYGAIKATRVTELLYRM
O1CAMPOS55 APHRVLATVYNGECRYSRNAVPNVRGDLQVLAQKVVRTLPTSFNYGAIKATRVTELLYRM
O1MANISA69
[AJ251477]
Cattle
challenge
virus
Capsid
Protein
61
N
N
N
K
K
K
K
139
A
A
A
P
P
P
P
163
S
S
S
P
P
P
S
57
P
R
R
P
P
P
P
137
G
V
V
V
V
V
V
VP2
Examination of the B-cell epitopes in the 1D region show perceptible differences
VP1
No changes were noticed in VP2 region
“New tools and challenges for progressive control”
Open Session of the EuFMD Research Group, Vienna (Austria) 29 September ‐ 1 October 2010
Appendix 30
Sequence identities of the O1 Campos viruses used in vaccine
production, cattle challenge test and neutralisation test
Capsid
Protein
Comparison of r1 values for protection against
heterologous challenge
r1 values
Protected
Not Protected
Total
% Protection
<0.19
2
2
4
50.00
A
0.20-0.39
2
1
3
66.67
A
A
0.40-0.70
5
4
9
55.56
A
A
A
0.71-1.00
4
3
7
57.14
A
A
A
>1
9
6
15
60.00
Amino acid
Position
O1CAMPOS58
[AJ320488]
BHK5
BHK6
Cattle
Passage 1
Cattle
Passage 2
Cattle
challenge
virus
Affected
cattle - 4167
Feet
Epithelium
4
A
A
A
V
V
V
13
T
T
A
A
A
97
G
G
A
A
156
V
V
A
A
VP1
No changes were noticed in VP4, VP2 and VP3 regions
Mean Protection
Comparison of serum antibody titres for
protection against heterologous challenge
≤ Titres
Based on O1 Manisa titres
Protected
Not Protected
Total
% Protection
0.61
1.00
0.00
1.00
100.00
0.91
0.00
1.00
1.00
1.21
0.00
2.00
2.00
1.51
1.00
5.00
1.81
7.00
6.00
2.11
4.00
2.00
2.41
4.00
2.71
2.00
3.01
1.00
57.87
Comparison of serum antibody titres for
protection against heterologous challenge
≤ Titres
Based on O1 Campos titres
Protected
Not Protected
Total
% Protection
0.61
1.00
1.00
2.00
50.00
0.00
0.91
0.00
1.00
1.00
0.00
0.00
1.21
2.00
1.00
3.00
66.67
6.00
16.67
1.51
2.00
7.00
9.00
22.22
13.00
53.85
1.81
4.00
5.00
9.00
44.44
6.00
66.67
2.11
2.00
2.00
4.00
50.00
1.00
5.00
80.00
2.41
5.00
1.00
6.00
83.33
1.00
3.00
66.67
2.71
3.00
0.00
3.00
100.00
0.00
1.00
100.00
3.01
1.00
0.00
1.00
100.00
Mean protection
52.63
Interpretations
Mean protection
52.63
Interpretations
• The two experiments produced almost similar results.
• Vaccine containing the highest payload of 60 µg could offer only partial
protection (75%) in Expt 1 where as in Expt II it was 100%.
• Post-challenge generalization of the disease was noticed in all but one
unvaccinated control animal.
• Vaccines containing the payload of 15 µg could offer only 50-60%
protection.
• FMD-NSP antibodies could be detected from all the control animals until
day 35 post challenge except in case of two animals in the homologous
challenge group.
• Hence the results confirm that there is only partial cross protection for O1
Campos in animals vaccinated with O1 Manisa
• Viral RNA/virus could be detected intermittently from the control animals
up to day 35 post-challenge in most of the animals.
• To achieve cross protection O1 Manisa pay load must be very high or
possibly a repeat vaccination with the optimal payload may offer protection
(to be confirmed by further studies)
• Two control animals each in homologous and heterologous challenge did
not show virus RNA/virus on day 35 post challenge.
“New tools and challenges for progressive control”
Open Session of the EuFMD Research Group, Vienna (Austria) 29 September ‐ 1 October 2010
Appendix 30
Interpretations
• NSP antibody response was noticed in most of the animals indicating virus
replication in both the experiments
• Virus was not isolated from the probang samples in any of the vaccine
groups.
• However, viral RNA could be detected from one animal in all the O1
Manisa challenged groups and one each in two O1 Campos challenged
groups (60 and 3.75 µg) by qRT-PCR.
• The Log PA50 values for serum antibody tires for O1 Manisa was 0.879
while that for O1 Campos was 1.739
• The PD50 values for antigen payload for O1 Manisa was 0.92 mcg/dose
while that for O1 Campos was 32.2 mcg/dose.
Thank you
Acknowledgements
"The research leading to these results have received
funding from the European Community's Seventh
Framework Programme (FP7/2007-2013) under grant
agreement n° 226556"
“New tools and challenges for progressive control”
Open Session of the EuFMD Research Group, Vienna (Austria) 29 September ‐ 1 October 2010