Vaccine efficacy of FMD virus -like particles like particles like particles like particles produced by the produced by the produced by the baculovirus expression system expression system expression (Van den Born)

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The European Commission for the Control of Foot-and-Mouth Disease (EuFMD), one of FAO’s oldest Commissions, came into being on the 12th June 1954, with the pledge of the sixth founding member state to the principles of a coordinated and common action against Foot-and-mouth Disease.

Government & Nonprofit

Vaccine efficacy of FMD virus-like particles
produced by the baculovirus expression system
Erwin van den Born
EuFMD 2018

2
Improving VLP stability
PROTECTIVE NON-PROTECTIVE

3
Improving VLP stability
VP2 (wt)
VP2-93Phe
Example: Stabilization of
the VP2-VP2 interface
VP2-93Cys
Not stabilized
Stabilized
Cysteine
bridge
Electrostatic
interactions

4
Optimization of VLP production
• High yield of VLPs important to get
• affordable vaccines
• multivalent vaccines
• How to improve the yield
• Optimization of the baculovirus expression system
• Stabilization of the capsids
• Yield of FMDV protein increased ~102 fold

5
Generation of stabilized VLP based on 4 serotypes
Preparation of VLPs
A O Asia1 SAT2
Inactivated
FMD virus
Empty
capsids

6
Efficacy of VLP vaccines in cattle
Serotype / vaccine VN titres ≥1.5log10 Protection
O VLP (stabilized) Yes Yes
O classic Yes Yes
A VLP (stabilized) Yes Yes
A classic Yes Yes
Asia1 VLP (stabilized) Yes Yes
Asia1 classic Yes Yes
SAT2 VLP (stabilized) Yes Trial scheduled
SAT2 classic Yes Trial scheduled
- 5-6 cattle per group
- VLP vaccine contains same adjuvant as MSD current vaccine (double oil emulsion)
- Homologous challenge at 3 wpv

7
Product development aspects
VLP vaccine
Cell and
virus seeds
(EA test)
Animal
safety and
efficacy
Product
registration
Vaccine
stability
Upstream &
downstream
processing
Batch
release test
(potency)

8
Large scale production of VLP
Insect
cells
Cell cultivation Baculovirus inoculation
and VLP expression
Harvest culture Inactivation of
recombinant
baculoviruses
(multivalent)
vaccine formulation
Baculo-
virus
Concentration

10
In vitro potency test
Benefits in vitro potency test compared to batch release test using animals:
- Cheaper
- Quicker (1 day vs weeks/months)
- Fullfils the 3R principles of animal testing (replacement, reduction, refinement)
- Less variation (risk of batch failure is reduced)
- No need for high containment
X µg of A strain
Y µg of O strain
Z µg of Asia1 strain
VN test A
VN test O
VN test Asia1
Current in vivo potency test:
Requirement for high
containment and cell-
adapted viruses

11
In vitro potency test
0.0
0.5
1.0
1.5
2.0
2.5
100 300 900 2700 81002430072900218700
OD450nm
Dilution
unheated
heated, 56°C
12S particles
146S particles
Options:
 HPLC
 ELISA
 Other?
PROTECTIVE NON-PROTECTIVE
Requirements of an in vitro potency test:
- Can differentiate between 146S and 12S
- Can differentiate between serotypes

12
• Virus-like particles can be produced by baculovirus expression
• Yield and stability of capsids can be significantly improved
• Technology works for at least serotypes A, O, Asia1 and SAT2
• To maximize the benefit of the VLP technology (i.e. no high
containment, quick response to market, affordable), an in vitro batch
release test is desirable.
Virus-like particles have the potential to be a commercially viable
alternative to conventional killed virus vaccines
Summary

13
Acknowledgements
Pirbright Institute, UK
Bryan Charleston
Claudine Porta
Eva Perez
Sophia Hodgson
University of Reading, UK
Ian Jones
Silvia Loureiro
MSD Animal Health, Netherlands
Ruud Segers
Amaya Serrano García
Alexandra Jiménez-Melsió
Holger Hönemann
University of Oxford, UK
Dave Stuart
Abhay Kotecha
Elizabeth Fry
Jingshan Ren

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LSD symposium - L. Pite - Combating lumpy skin disease in Albania

EuFMD

1) The first case of Lumpy Skin Disease (LSD) in Albania was identified in June 2016. From 2016-2017, over 3,500 outbreaks were reported across Albania with morbidity of 42% and mortality of 12%. 2) Surveillance efforts included laboratory testing of over 2,000 samples confirming 881 positive cases. Risk factors for spread included proximity to infected farms (under 5km), livestock movements over longer distances, and seasonal variations correlated with temperature and vector abundance. 3) Control efforts included an emergency vaccination program using live attenuated vaccine beginning in July 2016. Over 500,000 vaccine doses were administered. Modeling estimated vaccine effectiveness was 76.5-62.5% at reducing

LSD symposium - E. Tuppurainen - Update on global distribution of lumpy skin ...

EuFMD

LSD symposium - J. Chan - Lumpy skin disease in Hong Kong

EuFMD

Dr. Jason Chan presented on the outbreak of Lumpy Skin Disease (LSD) in feral cattle populations in Hong Kong from 2020-2021. The key points were: 1) The initial outbreak was reported in October 2020 across multiple country parks. Disease investigation found that 72% of cattle in one herd showed skin lesions and 84% were seropositive. 2) By March 2021, no new clinical cases were reported. Surveillance since found 14 juveniles seronegative, suggesting LSD may have disappeared due to lack of susceptible newborn cattle. 3) Continued clinical and serological surveillance is important since Hong Kong has a small teaching farm. No urgency exists currently to declare freedom

LSD symposium - N. Zainuddin - Indonesian experience on simultaneous LSD and ...

EuFMD

1) Lumpy skin disease was first reported in Indonesia in February 2022 in Riau Province, and has since spread to several other provinces, most recently to Central Java in August 2022. 2) As of February 2023, over 249,000 cattle have been vaccinated across 9 provinces as a control measure. Other control measures include movement restrictions, vector control, and educating farmers. 3) Key challenges to control efforts include the extensive animal farming system, illegal animal movement, limited number of vaccinators, and high workload from controlling both lumpy skin disease and foot-and-mouth disease. Recommended solutions include improving handling capacity, better border control, engaging other institutions to assist with vaccination

LSD symposium - R. Ainsworth - Lumpy skin disease (LSD) in Southeast Asia Mar...

EuFMD

Lumpy Skin Disease (LSD) is spreading through cattle movements in Southeast Asia. The document discusses how government policies around quarantine, compensation and corruption can accelerate the virus's spread by encouraging illicit cattle movements. It also notes that traditional smuggling routes go against the direction LSD has spread. The rapid transmission of LSD occurred during COVID border closures, and its direction of movement corresponds with prevailing winds rather than cattle trade routes. Government policies and wind patterns may be aiding the long-distance airborne spread of LSD across Southeast Asia.

LSD symposium - P. Malik - Lumpy skin disease experience from India

EuFMD

Lumpy Skin Disease (LSD) was first reported in India in 2019. It has since spread to 23 states and union territories, affecting over 3 million animals and causing over 185,000 deaths. The disease manifests as skin nodules and lesions on internal organs. Vaccination is a key control strategy, with over 87 million animals vaccinated to date using a goatpox vaccine. ICAR has also developed an indigenous LSD vaccine that is undergoing field trials and licensing. States are implementing control measures like quarantine, vaccination, vector control and public awareness campaigns to curb the spread and impact of LSD.

LSD symposium - G. Cattoli - Lessons from ten years of experience building me...

EuFMD

LSD symposium - H. Bergmann - Systemic review and expert ranking of potential...

EuFMD

LSD symposium - E. Klemen - Modes of transmission of lumpy skin disease

EuFMD

Indirect transmission, likely through blood-sucking flying insects, is the primary mode of transmission for lumpy skin disease virus. While direct contact can transmit the virus, studies have found no transmission between clinically infected and susceptible cattle housed together without vectors. Mathematical models also indicate indirect transmission alone can explain outbreak dynamics. The virus can spread over long distances, possibly aided by winds carrying infected vectors, though local spread is typically 10 km per week. Subclinical infections may transmit the virus but appear to play a minor role compared to clinical cases.

LSD symposium - A. Van Schalkwyk - Evolutionary changes between LSDV strains ...

EuFMD

This document summarizes the evolution of lumpy skin disease virus (LSDV) strains in South Africa over time. It discusses: 1) The spread of LSDV from northern Rhodesia (now Zambia) to South Africa in the 1940s and isolation of the first South African strain, Neethling, in 1957. 2) Genetic analysis showing Neethling strain from 1957 is the prototype for cluster 1.1 strains found only in South Africa until 1997, accumulating around 60 SNPs over 43 years. 3) The Kenyan strain from 1958 is the prototype for cluster 1.2, now the only LSDV observed in South Africa, accumulating around 48 SNPs over 22 years

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