Technical development of a novel vaccine vehicle for cattle pathogens
Lead Research Organisation:
University of Edinburgh
Department Name: Sch of Biological Sciences
Abstract
Trypanosoma theileri is a ubiquitous single celled organism present in cattle herds worldwide. Using knowledge of gene expression and protein trafficking in other trypanosomatid organisms, we have successfully developed T. theileri as a delivery system for vaccine antigens and therapeutics. Following recommendations from several Animal Health companies, we now aim to develop and demonstrate the potential of T. theileri as an effective and practical vaccine-vehicle, providing a clear route to commercialisation.
In a two-step development path, Phase 1 will focus on technical refinements and analysis of the viability of the vaccine-vehicle during storage and distribution. The flexibility of the system to express antigens from a range of pathogens will also be established. Phase 2 will entail confirmation of the in vivo efficacy of the refinements developed in Phase 1.
Combined the experiments will address key hurdles to the commercial development of this patented vaccine system.
In a two-step development path, Phase 1 will focus on technical refinements and analysis of the viability of the vaccine-vehicle during storage and distribution. The flexibility of the system to express antigens from a range of pathogens will also be established. Phase 2 will entail confirmation of the in vivo efficacy of the refinements developed in Phase 1.
Combined the experiments will address key hurdles to the commercial development of this patented vaccine system.
People |
ORCID iD |
| Keith Matthews (Principal Investigator) |
Publications
Contreras Garcia M
(2022)
Comparative Sensitivity and Specificity of the 7SL sRNA Diagnostic Test for Animal Trypanosomiasis.
in Frontiers in veterinary science
Garcia, M. C.
(2022)
Comparative sensitivity and specificity of the 7SL sRNA diagnostic test for Animal Trypanosomiasis
in Frontiers in Veterinary Science
Oldrieve GR
(2022)
The genomic basis of host and vector specificity in non-pathogenic trypanosomatids.
in Biology open
| Description | This project stated in October 2014. The experiments engineered Trypanosoma theileri to express several heterologous antigens from episomal vectors. Many constructs to optimise the system and ensure containment were developed; in particular the maintenance of the episomal plasmids was assessed in vitro using GFP reporter expression and maintenance of antibiotic resistance, which was encoded on the transfected plasmid. This demonstrated that plasmids were maintained in the absence of selection for around 3 months- but then lost; this represented a very good kinetic of expression suitable for in vivo vaccination and immune stimulation. Also, we have explored the cold chain stability of Trypanosoma theileri and have generated robust and key data required by industry to assess the practical application of our novel delivery vehicle. In 2016/2017 we carried out in vivo trials using episomal expression vectors, as well as integrative expression vectors in an auxotrophic mutant. The immunogenicity of the new vehicles has been analysed and compared with earlier studies. To date our evidence is that the alternative delivery routes using episomal vectors can stimulate immunity but this is not as good as had been previously seen with a construct integrated into the genome of the parasite. It is not clear why the immune stimulation was less effective but it might be related to the expression levels of the antigen. This can be improved in future by codon optimisation. The genome of the T. theileri vehicle has also been completed and analysed in comparison to pathogenic trypanosomes and this has revealed some of the unique biology of both parasites; for T. theileri this highlighted the adaptations for immune evasion and how these differ from T. brucei. Finally, as an extension to the funded work we developed methods to differentiate the parasite to its insect form because this is expected to show enhanced thermal stability compared with the bloodstream form parasites. We also adapted to culture and successfully transfected a related Trypanosomatidae of sheep, Trypanosoma melophagium, which might form the basis of a vaccine delivery vehicle for ovine pathgens- similar to the approach using T. theileri for cattle pathogens. |
| Exploitation Route | Our research programme aimed to evaluate and develop a novel system for the sustained delivery of an unlimited range of pathogen antigens specific to cattle. Being defined and characterised in terms of their immunogenicity, vaccinated and diseased animals are able to be identified and distinguished. Moreover, the technology proposed has economic advantages over the existing expensive and labour-intensive production of attenuated vaccines. This offers clear advantages to the agricultural and animal healthcare industry at large. The proposal developed early-stage work, with substantial commitment from an Industrial Partner, to demonstrate utility and provide the required scientific validation to stimulate further development in an industrial setting with the partner company. The proposal fulfilled the strategic remit of the animal sciences panel with respect to: The 'control of infectious diseases', (priority area) comprising 'identification of protective antigens', 'vaccinology' and 'improved antigen delivery systems'. Strategic plan objectives for 'Sustainable Agriculture' comprise 'host/pathogen interactions', 'animal welfare' and the 'control of animal diseases'. In 2016/2017 we successfully secured further funding based on the concepts developed in this proposal. One grant was for development of an equivalent vaccination system using sheep trypanosomatids (Royal Society Challenge award), based on preliminary work carried out in this proposal. Another was to explore the viability of the current system to vaccinate against major zoonotic infections that pose risk for human epidemics (Innovate UK). Finally, application of the approach to combat East Coast Fever has been pursued through the award of funding through the Bill and Melinda Gates foundation to assess the delivery vehicle for effective immunisation against that pathogen. This new funding is currently underway. There is potential for further ongoing funding from the Bill and Melinda Gates foundation to support this ongoing work. We have also secured commercial interest on the Trypanosoma melophagium platform for vaccination against sheep pathogens. We negotiated contributory support from Roslin Technologies Etc and the BBSRC impact accelerator funds to pursue proof of concept work around this activity. This was successful with a total award of £118000 provided, of which £88000 was from Roslin Technologies. The work is currently ongoing. |
| Sectors | Agriculture, Food and Drink,Education,Manufacturing, including Industrial Biotechology |
| Description | The work has been progressed to European patent filing, and this was granted in early 2017. A US patent was also awarded in 2018. Follow on work has also been funded in collaboration with Roslin Technologies via BBSRC IAA funding (totalling £118000) and this work is currently ongoing. |
| First Year Of Impact | 2017 |
| Sector | Agriculture, Food and Drink |
| Impact Types | Economic |
| Description | BBSRC IAA |
| Amount | £118,956 (GBP) |
| Funding ID | PIII078 |
| Organisation | University of Edinburgh |
| Sector | Academic/University |
| Country | United Kingdom |
| Start | 04/2020 |
| End | 04/2021 |
| Description | GCRF Challenge award |
| Amount | £117,261 (GBP) |
| Funding ID | CH160034 |
| Organisation | The Royal Society |
| Sector | Charity/Non Profit |
| Country | United Kingdom |
| Start | 12/2016 |
| End | 10/2018 |
| Description | SBRI Vaccines for Global Epidemics - Preclinical Stage 1 |
| Amount | £334,000 (GBP) |
| Funding ID | 86235-544135 |
| Organisation | Innovate UK |
| Sector | Public |
| Country | United Kingdom |
| Start | 04/2017 |
| End | 04/2018 |
| Description | Supporting Evidence Based Interventions-Evaluation of a trypanosomatid vaccine vehicle to combat livestock infections in LMIC |
| Amount | £191,361 (GBP) |
| Organisation | Bill and Melinda Gates Foundation |
| Sector | Charity/Non Profit |
| Country | United States |
| Start | 04/2019 |
| End | 09/2020 |
| Title | Growth and transfection of Trypanosoma melophagium |
| Description | As a complement to our work with the cattle trypanosomatid Trypanosoma theileri, we also isolated and adapted to culture a line of the sheep trypanosomatid, Trypanosoma melophagium. These were solvated from feral Soay sheep on St Kilda and adapted to in vitro culture. methods for their genetic transformation were then derived. |
| Type Of Material | Cell line |
| Year Produced | 2017 |
| Provided To Others? | No |
| Impact | Trypanosoma melophagium is being developed as a vaccination platform for sheep. This is under development. |
| Title | Trypanosoma theileri genome sequence |
| Description | Genome sequence |
| Type Of Material | Database/Collection of data |
| Year Produced | 2018 |
| Provided To Others? | Yes |
| Impact | Permits comparative genomic analysis with other kinetoplastid parasites |
| URL | https://www.ncbi.nlm.nih.gov/bioproject/PRJNA273795 |
| Description | Collaboration with Moredun |
| Organisation | Moredun Research Institute |
| Country | United Kingdom |
| Sector | Charity/Non Profit |
| PI Contribution | Provision of transgenic T theileri for the infection of cattle in a contained facility to evaluate immune responses generated toe expressed antigens |
| Collaborator Contribution | Provision of a contained cattle facility |
| Impact | See publication linked to this output |
| Start Year | 2009 |
| Description | Collaboration with Roslin Technologies Ltd |
| Organisation | Roslin Technologies |
| Country | United Kingdom |
| Sector | Private |
| PI Contribution | Work based on our Trypanosoma theileri studies have been redirected toward vaccine deployment for sheep using Trypanosoma melophagium. This is being evaluated via BBSRC impact accelerator support for commercial potential. The experimental work in 2022 demonstrated that an effective immune response was not generated to the antigen expressed by Trypanosoma melophagium and the work was stopped. |
| Collaborator Contribution | Roslin Technologies Ltd have provide considerable support (approximately £80,000) to match BBBSRC impact accelerator research funding. Within this they have funded a market assessment report on the commercial potential of the technology. |
| Impact | A publication exploring the hoist and vector specificity of trypanosomatid parasites in sheep and cattle was generated: The genomic basis of host and vector specificity in non-pathogenic trypanosomatids. Oldrieve GR, Malacart B, López-Vidal J, Matthews KR. Biol Open. 2022 Apr 15;11(4):bio059237. doi: 10.1242/bio.059237. Epub 2022 May 3. |
| Start Year | 2020 |
| Description | Collaboration with researchers at Roslin Institute |
| Organisation | University of Edinburgh |
| Department | The Roslin Institute |
| Country | United Kingdom |
| Sector | Academic/University |
| PI Contribution | We expressed antigens of the cattle Parasite Trypanosoma congolense in Trypanosoma theileri and this was used as the basis of experiments to evaluate the ability of these T congolense antigens to generate immune responses in cattle. |
| Collaborator Contribution | Antigen sequences were provided to generate the recombinant T. theileri cell lines and these are being evaluated by them for immunogenicity in cattle |
| Impact | A funding application has been submitted to the Bill and Melinda Gates foundation to progress development of this vaccination approach |
| Start Year | 2016 |
| Description | Intervet collaboration |
| Organisation | Intervet-Schering Plough |
| Country | United States |
| Sector | Private |
| PI Contribution | We evaluated a novel vaccine vehicle for the delivery of antigens to cattle. We expressed an antigen provided by Intervet (after expression optimisation of the vaccine delivery vehicle) and tested its immunogenicity in cattle. |
| Collaborator Contribution | Intervet provided technical guidance, reagents and IP protection support. They also provided a cash contribution to the award as required by the Industrial Partnership scheme. |
| Impact | Publication Mott, G. A.; Wilson, R.; Fernando, A.; Robinson, A.; MacGregor, P.; Kennedy, D.; Schaap, D.; Matthews, J. B. and Matthews, K. R. (2011) Targetting cattle-borne zoonoses and cattle pathogens using a novel trypanosomatid-based delivery system. PLoS Pathogens, 7(10):e1002340. Outreach Wellcome Trust blog: http://blog.wellcome.ac.uk/2012/02/14/neglected-tropical-diseases-working-with-animals/ BBSRC news feature: http://www.bbsrc.ac.uk/news/food-security/2011/111028-pr-cattle-parasite-vaccine.aspx Veterinary record news report: http://veterinaryrecord.bmj.com/content/169/21/541.1.extract |
| Start Year | 2009 |
| Description | collaboration with the 'Supporting Evidence based Interventions' programme at the University of Edinburgh |
| Organisation | University of Edinburgh |
| Country | United Kingdom |
| Sector | Academic/University |
| PI Contribution | Our vaccine vehicle technology is being use din a collaboration with the Supporting Evidence Based interventions programme at the University of Edinburgh to investigate the potential of using the cattle vaccination approach to target thjileria parasites in Africa |
| Collaborator Contribution | The SEBI programme seeks to develop novel technologies to address issues of livestock productivity in sub Saharan Africa. It is funded by the Bill and Melinda Gates foundation. |
| Impact | The work is ongoing |
| Start Year | 2018 |
| Title | RECOMBINANT TRYPANOSOMA THEILERI PARASITE |
| Description | Patent for the production of recombinant Trypanosoma theileri for the purposes of vaccine delivery |
| IP Reference | EP2598163 |
| Protection | Patent granted |
| Year Protection Granted | 2013 |
| Licensed | No |
| Impact | None |
| Title | RECOMBINANT TRYPANOSOMA THEILERI PARASITE |
| Description | The present invention relates to the field of the veterinary medicine of bovine animals. In particular the invention relates to a recombinant Trypanosoma theileri parasite, preferably comprising a heterologous nucleic acid sequence that is capable of encoding a protein for instance an antigen, a cytokine, a hormone, an antimicrobial protein, or an antibody. Also disclosed are uses of and methods for making and using the recombinant T. theileri parasite in medical or non-curative treatments; in particular as a sustained delivery vector for proteins to bovine animals, e.g. as a vaccine. |
| IP Reference | WO2012013939 |
| Protection | Patent application published |
| Year Protection Granted | 2012 |
| Licensed | No |
| Impact | The patent application is under review in the US at present. Development work is being supported by a BBBSRC Super Follow on fund application |
| Description | FAO/IAEA Symposium on Sustainable Improvement of Animal Production and Health |
| Form Of Engagement Activity | A talk or presentation |
| Part Of Official Scheme? | No |
| Geographic Reach | International |
| Primary Audience | Professional Practitioners |
| Results and Impact | Postdoctoral research Javier Lopez Vidal will present data on the use of transgenic Trypanosoma theileri for vaccination against livestock diseases (meeting is scheduled for June 2021) |
| Year(s) Of Engagement Activity | 2021 |
| URL | https://www.iaea.org/events/aphs2021 |
| Description | vaccination for trypanosomatid parasites discussion group |
| Form Of Engagement Activity | Participation in an activity, workshop or similar |
| Part Of Official Scheme? | No |
| Geographic Reach | National |
| Primary Audience | Professional Practitioners |
| Results and Impact | Work shop on vaccination for trypanosomatid parasites; held at the Veterinary School, University of Edinburgh |
| Year(s) Of Engagement Activity | 2019 |