Studentship: The inflammatory response to Bluetongue virus and vector insect saliva in ovine and bovine immune and skin cells
Lead Research Organisation:
THE PIRBRIGHT INSTITUTE
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
Bluetongue virus (BTV) is the aetiological agent of an economically important disease of ruminants (bluetongue disease). Severe clinical signs develop mainly in sheep while infected cattle remain largely asymptomatic. The reason for the difference in disease manifestation is not fully understood, however different immune responses to BTV may play a vital role.
BTV is transmitted between its ruminant hosts almost exclusively by Culicoides biting
midges. Insect saliva proteins can elicit strong inflammatory reaction in mammalian skin and it has become evident that many arboviruses benefit from co-inoculation with insect saliva.
A method has been developed to collect Culicoides saliva proteins enabling the design of in vitro assays and proteomic analysis.
The overall aim of this PhD project is to investigate the inflammatory response of certain immune and skin cells towards BTV infections in the presence and absence of Culicoides saliva in vitro and compare the immune response of ovine and bovine host cells.
This project will investigate the following hypothesises:
1. The inflammatory response of certain (primary) immune and skin cells towards BTV
infections differs between bovine and ovine derived cells
2. Culicoides saliva elicits an inflammatory response in ovine and bovine immune and skin cells that influences BTV replication
3. Individual saliva proteins of Culicoides elicit specific inflammatory responses in immune cells and the saliva protein responsible can be identified
This multidisciplinary project will utilise a combination of techniques e.g. cell culture, virus infection and titration, confocal microscopy, real-time PCR, Proteomics, ELISA, flow cytometry and Culicoides saliva collection. Methods to fractionate Culicoides saliva while maintaining the biological activity will be developed. The successful candidate will acquire expertise in immunology, virology and proteomics
BTV is transmitted between its ruminant hosts almost exclusively by Culicoides biting
midges. Insect saliva proteins can elicit strong inflammatory reaction in mammalian skin and it has become evident that many arboviruses benefit from co-inoculation with insect saliva.
A method has been developed to collect Culicoides saliva proteins enabling the design of in vitro assays and proteomic analysis.
The overall aim of this PhD project is to investigate the inflammatory response of certain immune and skin cells towards BTV infections in the presence and absence of Culicoides saliva in vitro and compare the immune response of ovine and bovine host cells.
This project will investigate the following hypothesises:
1. The inflammatory response of certain (primary) immune and skin cells towards BTV
infections differs between bovine and ovine derived cells
2. Culicoides saliva elicits an inflammatory response in ovine and bovine immune and skin cells that influences BTV replication
3. Individual saliva proteins of Culicoides elicit specific inflammatory responses in immune cells and the saliva protein responsible can be identified
This multidisciplinary project will utilise a combination of techniques e.g. cell culture, virus infection and titration, confocal microscopy, real-time PCR, Proteomics, ELISA, flow cytometry and Culicoides saliva collection. Methods to fractionate Culicoides saliva while maintaining the biological activity will be developed. The successful candidate will acquire expertise in immunology, virology and proteomics
Planned Impact
unavailable
Publications
Darpel KE
(2016)
Bluetongue - an ever present threat to UK ruminants?
in Cattle Practise
Stevens L
(2019)
A low-passage insect-cell isolate of bluetongue virus uses a macropinocytosis-like entry pathway to infect natural target cells derived from the bovine host
in Journal of General Virology
| Description | Bluetongue virus (BTV) replicates to higher titres in cultured bovine peripheral blood mononuclear cells (PBMCs) in the presence of saliva from its insect vector Culicoides biting midges. A higher percentage of bovine monocytes within cultured PBMCs express BTV structural and non-structural proteins in the presence of Culicoides saliva. Infection enhancement effects are only detectable if the insect saliva is added at the same time as the virus or in the first few hours post infection suggesting an effect on virus entry or replication initiation. Cytokines produced by PBMCs following BTV infection are modified in the presence of Culicoides saliva. However individual animals seem to vary in specific responses assessed Overall these results further demonstrate that the saliva of its Culicoides biting midge vector has a significant impact on the infectivity and replication of bluetongue virus in natural host cells. |
| Exploitation Route | Culicoides spp. saliva contains numerous bioactive proteins many of them of yet unknown function. By further elucidating the effect of insect saliva on cellular innate immune responses and blood coagulation it might be possible to assign observed functions to specific proteins which could be exploited for other medical use. Observed impact of insect vector saliva on the infectivity and immune response to co-transmitted virulence might allow the design of better control measures to disrupt virus transmission including better vaccine design for insect transmitted viruses. The obtained results also further strengthen the argument that in-vivo studies investigating arbovirus pathogenesis, dissemination and transmission as well as host anti-viral responses need to consider the arthropod vector as a key part of the transmission cycle which will also influence infection outcome of the vertebrate host. |
| Sectors | Agriculture Food and Drink Pharmaceuticals and Medical Biotechnology |
| Description | Salivary proteins from Culicoides biting midges collected for this and other projects will be supplied to a company for inclusion in their veterinary allergy tests. Potential to widen the collection and usage of insect saliva to further health diagnostic areas |
| First Year Of Impact | 2016 |
| Sector | Agriculture, Food and Drink |
| Impact Types | Economic |
| Description | BBSRC responsive mode grant |
| Amount | £367,000 (GBP) |
| Funding ID | BB/P006841/1 |
| Organisation | Biotechnology and Biological Sciences Research Council (BBSRC) |
| Sector | Public |
| Country | United Kingdom |
| Start | 09/2017 |
| End | 03/2019 |
| Description | Pirbright Institute PhD studentship |
| Amount | £100,000 (GBP) |
| Organisation | The Pirbright Institute |
| Sector | Academic/University |
| Country | United Kingdom |
| Start | 09/2019 |
| End | 09/2023 |
| Description | Workpackage within a Horzion 20:20 grant |
| Amount | € 6,000,000 (EUR) |
| Funding ID | EU project 727393 Horizon 20:20 PALE-Blue |
| Organisation | European Union |
| Sector | Public |
| Country | European Union (EU) |
| Start | 05/2017 |
| End | 11/2021 |
| Description | Collaboration studentship BTV |
| Organisation | University of Surrey |
| Department | Faculty of Health and Medical Sciences |
| Country | United Kingdom |
| Sector | Academic/University |
| PI Contribution | Collaborative studentship with virologist at the University of Surrey, student registered at the University |
| Collaborator Contribution | Intellectual input, training of student |
| Impact | Thesis http://epubs.surrey.ac.uk/845643/ |
| Start Year | 2013 |
| Description | Microbial products in insect saliva and effects on virus infection enhancement |
| Organisation | Boston University School of Medicine |
| Country | United States |
| Sector | Academic/University |
| PI Contribution | Sharing of results on the discovery of bacterial LPS in Culicoides saliva and its infection enhancing effect on BTV in bovine monocytes. Discussion of future work and comparison of the results to other insect transmitted viruses including those of high importance to humans |
| Collaborator Contribution | Sharing of data from their portfolio on mosquito saliva effects on virus infectivity and replication. Comparison of methodology and discussion of future collaborations |
| Impact | On-going sharing of results and discussion of future experiments - collaborator left academia to work in industry. Contact still remains. |
| Start Year | 2018 |
| Description | Attandence of the BBSRC cross institute microbiome workshop (7th and 8th January 2019) |
| 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 | Attendance of the BBSRC cross institute microbiome workshop (7th and 8th January 2019) by postdoctoral scientist - discussion of future collaborations and links |
| Year(s) Of Engagement Activity | 2019 |
| Description | Epizone, Berlin - invited speaker |
| Form Of Engagement Activity | A talk or presentation |
| Part Of Official Scheme? | No |
| Geographic Reach | International |
| Primary Audience | Postgraduate students |
| Results and Impact | Invited speaker on "Orbiviruses on the move" at the EPIZONE conference in Berlin 2019, audience mixture of veterinary virologists, early career researchers and veterinary policy makers |
| Year(s) Of Engagement Activity | 2019 |
| Description | Hampshire sheep discussion forum |
| Form Of Engagement Activity | A talk or presentation |
| Part Of Official Scheme? | No |
| Geographic Reach | Regional |
| Primary Audience | Industry/Business |
| Results and Impact | Presented and then led a discussion group on bluetongue virus emergence, current research and potential control measures to farmers, veterinarians and interested member of the public |
| Year(s) Of Engagement Activity | 2016 |
| Description | Invited speaker VECTOR-BORNE DISEASES (VBD) IN THE UK - BIENNIAL MEETING 2018 John Innes Centre Norwich |
| Form Of Engagement Activity | A talk or presentation |
| Part Of Official Scheme? | No |
| Geographic Reach | National |
| Primary Audience | Professional Practitioners |
| Results and Impact | Invited speaker on "The Host-vector-pathogen Interface of Culicoides-borne Bluetongue Virus" at the VECTOR-BORNE DISEASES (VBD) IN THE UK - BIENNIAL MEETING 2018 John Innes Centre Norwich 4th-5th December 2018 |
| Year(s) Of Engagement Activity | 2018 |
| Description | Microbiology Society Annual conferences- invited speaker |
| Form Of Engagement Activity | A talk or presentation |
| Part Of Official Scheme? | No |
| Geographic Reach | International |
| Primary Audience | Other audiences |
| Results and Impact | Scientific talk on successful cross-species transmission of vector-borne viruses |
| Year(s) Of Engagement Activity | 2019 |
| Description | Participation in School outreach event by student |
| Form Of Engagement Activity | Participation in an activity, workshop or similar |
| Part Of Official Scheme? | No |
| Geographic Reach | Local |
| Primary Audience | Schools |
| Results and Impact | Participation in outreach activity organised as a bug hunt at local primary school by student on the grant |
| Year(s) Of Engagement Activity | 2017 |
| Description | Presentation of project results at the 13th International dsRNA Virus Symposium 24th-28th of September 2018 |
| Form Of Engagement Activity | A talk or presentation |
| Part Of Official Scheme? | No |
| Geographic Reach | International |
| Primary Audience | Professional Practitioners |
| Results and Impact | Poster and Oral presentations included key results of the studentship project |
| Year(s) Of Engagement Activity | 2018 |
| Description | Yearly flow cytometry training (2 days/ year) on ruminant inat eimmune cell recognition to MSc student by posgraduate student (LC) on this studentship |
| Form Of Engagement Activity | Participation in an activity, workshop or similar |
| Part Of Official Scheme? | No |
| Geographic Reach | Local |
| Primary Audience | Postgraduate students |
| Results and Impact | To enable postgraduate students to utilise and understand flow cytometry in immunology research an innate cellular immune responses |
| Year(s) Of Engagement Activity | 2014,2015,2016 |