Development of Novel, Highly Protective Foot and Mouth Disease Virus vaccines in conjunction with the Global FMDV Research Alliance
Lead Research Organisation:
John Innes Centre
Department Name: UNLISTED
Abstract
Abstracts are not currently available in GtR for all funded research. This is normally because the abstract was not required at the time of proposal submission, but may be because it included sensitive information such as personal details.
Technical Summary
The overall aim of the project is to perform small scale pilot studies to determine the suitability of plant-based vector systems for the expression of assembled capsids of foot-and-mouth disease virus (FMDV). Suitable FMDV sequences for subcloning into plant-based expression systems will be provided by IAH-P or Reading University. Using Hepatitis B core antigen (HBcAg) we have already established that virus-like particles (VLPs) can be produced and assembled in plants using vectors based on both full-length and deleted versions of Cowpea mosaic virus (CPMV) RNA-2. The advantage of the deleted version of RNA-2 (delRNA-2) is that it can tolerate large inserts and no longer has the capacity for unwanted spread in the environment. For this reason we will utilise the delRNA-2 approach for this project.
To achieve expression of assembled of FMDV capsids, sequencing encoding the P1 region of various FMDV serotypes will be inserted into delRNA-2. To achieve processing, the enzyme responsible for proteolytic cleavage, 3CPro, will be provided either in cis (i.e. fused to the P1 sequence) or in trans (on a separate construct). We will initially concentrate on serotype A as a suitable construct is available at the start of the project and serotype A capsids are known to be particularly stable. The P1 construct with and without 3CPro will be agroinfiltrated into leaves and the expression and processing of P1 into the mature capsid proteins will be monitored immunologically. The ability of the cleaved P1 proteins to assemble into VLPs will be assessed by a number of criteria (electron microscopy, sucrose gradients) and there ability to raise an immune response in animals will be determined in collaboration with IAH.
To achieve expression of assembled of FMDV capsids, sequencing encoding the P1 region of various FMDV serotypes will be inserted into delRNA-2. To achieve processing, the enzyme responsible for proteolytic cleavage, 3CPro, will be provided either in cis (i.e. fused to the P1 sequence) or in trans (on a separate construct). We will initially concentrate on serotype A as a suitable construct is available at the start of the project and serotype A capsids are known to be particularly stable. The P1 construct with and without 3CPro will be agroinfiltrated into leaves and the expression and processing of P1 into the mature capsid proteins will be monitored immunologically. The ability of the cleaved P1 proteins to assemble into VLPs will be assessed by a number of criteria (electron microscopy, sucrose gradients) and there ability to raise an immune response in animals will be determined in collaboration with IAH.
Planned Impact
unavailable
Organisations
People |
ORCID iD |
| George Lomonossoff (Principal Investigator) |
Publications
Madi M
(2015)
Development of a non-infectious encapsidated positive control RNA for molecular assays to detect foot-and-mouth disease virus
in Journal of Virological Methods
Montague NP
(2011)
Recent advances of cowpea mosaic virus-based particle technology.
in Human vaccines
Porta C
(2013)
Efficient production of foot-and-mouth disease virus empty capsids in insect cells following down regulation of 3C protease activity.
in Journal of virological methods
| Description | This work has demonstrated that the co-expression of the P1 coat protein precursor of FMDV and the 3C proteinase necessary for its processing can result in the correct processing of the precursor. To achieve this it was necessary to develop methods for the down-regulation of 3C expression as this is toxic to most cells. By deploying these methods it was ultimately demonstrated that VLPs can be produced in several expression systems. |
| Exploitation Route | The methods developed for controlling expression levels of the 3C proteinase are generic and can be applied to any situation in which a potentially toxic protein is being expressed. An immediate potential application is to the production of VLPs of other picornaviruses such as poliovirus. |
| Sectors | Agriculture Food and Drink Manufacturing including Industrial Biotechology Pharmaceuticals and Medical Biotechnology |
| Description | The overall aim of this collaborative project was to develop novel vaccines against Foot-and-mouth disease virus (FMDV) based on non-infectious virus-like particles (VLPs). A variety of expression systems, including plants, were investigated as production platforms. It was shown that VLPs could be produced as long as the toxicity of the 3C proteinase was modulated by down-regulating its expression. This work has paved the way to the development of next-generation FMDV vaccines. |
| Sector | Agriculture, Food and Drink,Pharmaceuticals and Medical Biotechnology |
| Impact Types | Economic |