Elucidating local and systemic immune responses of livestock to arbovirus infections and arthropod saliva.

Lead Research Organisation: The Pirbright Institute
Department Name: UNLISTED

Abstract

This interdisciplinary research project will study the immune response and pathogenesis of arboviral infections in their mammalian hosts while additionally addressing the effect of vector arthropod saliva. The majority of studies investigating the immune response towards arboviruses and vector arthropod saliva will be carried out using their natural mammalian host, either in vivo or by establishing primary and/or ex vivo cell and organ culture systems. A major aspect of the in-vivo studies will be the cannulation of superficial lymphatic vessels in cattle and sheep to compare the inflammatory and migratory cell response in the collected lymph between infected and non-infected as well as arthropod exposed and non-exposed animals. The project will also investigate if certain aspects of the innate and systemic immune response contribute to the development of disease in mammalian hosts, rather than conferring protection.

Initial studies will focus on the immune response of ruminants towards bluetongue virus infection and Culicoides spp. blood-feeding. Innate and systemic immune responses in ruminants towards Culicoides spp, can later be compared to responses of hosts towards other haematophagous arthropod vectors (e.g. ticks and mosquitoes) as well as additional arboviruses. The in-vitro work studying the immune response of skin cells and endothelial cells has the additional benefit of being transferable to include equine derived cells, therefore allowing research into important horse disease or in the wider future important zoonotic arboviruses such as West Nile virus.

Publications

10 25 50
 
Description Additionally work funded by this award contributed to implementing a highly relevant in vivo animal study design where BTV is transmitted to the natural ruminant host via the blood-feeding of infected Culicoides biting midges. Combining infection studies in the natural host with insect transmission of the virus mimics natural arbovirus infection more closely and realistically than needle inoculations.

An ELISA was developed to detect antibodies (immunoglobulins G, M and E) against Culicoides saliva proteins to analyse the immune response of cattle and horses to Culicoides blood-feeding. We have further analysed ruminant and horse serum by western blot to identify the specific immunogenic salivary proteins and while different serum anti-saliva IgG profiles are detectable certain proteins are dominantly recognised.

The methodology of saliva collection was further transferred to Aedes aegypti mosquitoes - allowing to extend current research of the influence of arthropod vector saliva to mosquito-borne viruses. We previously demonstrated that saliva of the vector Culicoides sonorensis efficiently cleaves the outer coat protein of BTV, thereby changing its infectivity. In contrast Ae. aegypti saliva was not able to modify BTV proteins, suggesting that the interaction of vector saliva - arbovirus might reflect a selective relationship.

Furthermore, we have established primary bovine cell culture models (monocytes, endothelial cells) as well as organ model ex vivo and demonstrated successful infection with BTV allowing the detection of BTV infected cells in model tissue sections as well as detection of progeny virus in organ culture supernatant.
Exploitation Route The current studies investigating the effect of Culicoides saliva on co-transmitted viruses can also be transferred to research on other insect vector- arboviruses systems. Successfully transferring the methodology of saliva collection to Aedes mosquitoes will result in the availability of sufficient amounts of mosquito saliva for in-vitro studies on mosquito borne viruses.

Furthermore being able to carry out in vivo studies using the natural host, vectors and virus complexes will provide a unique opportunity to compare results with those generated in models (mostly rodents) for animal and human arthropod-borne pathogen research. Other international research team have already implemented bluetongue virus pathogenesis studies in the context of Culicoides blood-feeding exposure to assess the impact on BTV virulence and infectivity
Sectors Agriculture, Food and Drink

 
Description Findings that some BTV strains ( such as BTV-26) can be transmitted by contact and others by oral and mechanical transmission routes will influence BTV control strategies, inform veterinary practitioners and impact on the design of BTV surveillance and diagnostic strategies
First Year Of Impact 2014
Sector Agriculture, Food and Drink
Impact Types Economic

 
Description Bluetongue virus information campaign (JAB campaign led by National farmers Union)
Geographic Reach Local/Municipal/Regional 
Policy Influence Type Influenced training of practitioners or researchers
Impact The "Joint action against bluetongue" campaign of the National Farmers Union provided Veterinarians and Farmers with the opportunity to make an informed decision on BTV vaccination in accordance with the latest research following the re-emergence of BTV-8 in France and the subsequent availability of commercial BTV-8 vaccine. Preventing a Bluetongue virus outbreak in the UK will greatly benefit the national agriculture sector
 
Description BBSRC The Pirbright Institute PhD studentship
Amount £48,000 (GBP)
Funding ID BBS/E/I/00001728 
Organisation Biotechnology and Biological Sciences Research Council (BBSRC) 
Sector Public
Country United Kingdom
Start 10/2013 
End 09/2017
 
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 Collaborative studentship PPRV 
Organisation University of Surrey
Department School of Veterinary Medicine
Country United Kingdom 
Sector Academic/University 
PI Contribution Practical laboratory work and in vivo animal studies carried out
Collaborator Contribution Intellectual input in scientific remit of studentship, training of student, use of facilities and lab
Impact DOI: 10.1128/JVI.01471-18 Thesis: The immune response to live, attenuated peste des petits ruminants virus vaccines
Start Year 2014
 
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 Pathology of bovine skin post Culicoides midge blood-feeding 
Organisation University of Surrey
Department School of Veterinary Medicine
Country United Kingdom 
Sector Academic/University 
PI Contribution Collection of skin samples from cattle responding with local inflammation to Culicoides biting midges blood feeding, sending of materials
Collaborator Contribution Processing of samples, histopathological analysis, sharing of results
Impact Further identification of cellular and histological local changes in ruminant skin to Culicoides blood-feeing which will inform pathogenesis of co-transmitted arboviruses such as BTV
Start Year 2017
 
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 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 Video podcast Understanding Animal Research 
Form Of Engagement Activity A broadcast e.g. TV/radio/film/podcast (other than news/press)
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Public/other audiences
Results and Impact Participated in filming and interviewing for a labanimaltour where 4 Research Institutes openly discuss the use of animals in research. Currently already more than 1000 views of the Bluetongue virus related video either on youtube or facebook
Year(s) Of Engagement Activity 2016,2017
URL http://www.labanimaltour.org
 
Description Video potcasts on youtube for BTV clinical signs 
Form Of Engagement Activity A broadcast e.g. TV/radio/film/podcast (other than news/press)
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Professional Practitioners
Results and Impact Produced a youtube video on the clinical signs of bluetongue virus (an arbovirus of ruminants) together with colleagues from The Pirbright Institute and Animal and Plant Health Agency

3000 views so far
Year(s) Of Engagement Activity 2016
URL https://www.youtube.com/watch?v=fwJ8zAxiLOE