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

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
 
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 10/2017 
End 03/2019
 
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 07/2017 
End 07/2021
 
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
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
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 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 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