Use of reverse genetics systems to study avian viruses responsible for endemic diseases and the development of novel vaccine strategies

Lead Research Organisation: The Pirbright Institute
Department Name: UNLISTED

Abstract

This project will use reverse genetics systems to study the role of genes involved in viruses responsible for causing endemic diseases in poultry. This will involve studying the role of the genes in the biology of the viruses, their transmission and in pathogenicity. The work will allow the rational development of molecularly defined vaccines and generation of diagnostic reagents. Other aspects of the work will involve the identification interactions between viral and host proteins to help understand their role in pathogenicity. The ultimate outcomes of the studies will provide information for the development of new generations of vaccines against avian viruses using novel approaches.

Publications

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Bickerton E (2014) Cellular tropism of the infectious bronchitis virus spike glycoprotein in 8th International Symposium on Avian Corona- and Pneumoviruses and complicating pathogens 2nd Annual Meeting of Cost Action FA1207, Rauischholzhausen, Germany

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Bickerton E (2017) Reverse Genetics System for the Avian Coronavirus Infectious Bronchitis Virus. in Methods in molecular biology (Clifton, N.J.)

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Britton P (2012) Recombinant vaccines against infectious bronchitis in Avian Pathology Newsletter

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Dent SD (2015) The proteome of the infectious bronchitis virus Beau-R virion. in The Journal of general virology

 
Description We have used the reverse genetic system we developed for the pathogenic M41 strain of avian coronavirus infectious bronchitis virus (IBV) to investigate the role of the spike glycoprotein in tropism and pathogenicity. To do this, we generated several recombinant IBVs expressing the whole spike glycoprotein or a chimaeric spike glycoproteins from different strains of IBV. We assessed replication of the recombinant strains in vitro in primary chicken kidney cells and in Vero cells, a continuous cell line and confirmed that the extended tropism of the Beaudette strain of IBV in cell culture is conferred by the S2 subunit of the spike glycoprotein. We also assessed replication of one recombinant IBV in vivo.
Exploitation Route Knowledge about how recombinant IBVs with chimaeric spike glycoproteins replicate in cell culture will be of use in the rational design of next generation vaccines against IBV. This may aid the design of vaccines that can be propagated in cell culture rather than in embryonated eggs, an important consideration under the 3Rs. The findings of this work has contributed to the application and funding of a BBSRC Follow-on Fund grant, which will continue to develop some aspects of this research.
Sectors Agriculture, Food and Drink,Pharmaceuticals and Medical Biotechnology

 
Description The findings have been used in the design of new vaccine candidates against avian coronavirus, infectious bronchitis virus (IBV). These vaccines are being evaluated in collaboration with industrial partners. Funding has been awarded from BBSRC to continue some aspects of the research. Vaccines that can be produced in cell culture rather than in embryonated eggs would be very beneficial for the poultry industry.
First Year Of Impact 2016
Sector Agriculture, Food and Drink,Pharmaceuticals and Medical Biotechnology
Impact Types Societal

 
Description Contribution to a POSTnote on 'Reducing UK Antibiotic Use in Animals'
Geographic Reach National 
Policy Influence Type Participation in a advisory committee
 
Description Training MSc students
Geographic Reach Local/Municipal/Regional 
Policy Influence Type Influenced training of practitioners or researchers
Impact Delivered a new training session to MSc students teaching viral quantification assays. This lead to a greater understanding of laboratory techniques and academic research.
 
Description BBSRC responsive mode link award
Amount £1,382,000 (GBP)
Funding ID BB/P019137/1 
Organisation Biotechnology and Biological Sciences Research Council (BBSRC) 
Sector Public
Country United Kingdom
Start 02/2018 
End 01/2021
 
Description British Egg Marketing Board PhD studentship
Amount £76,000 (GBP)
Organisation British Egg Marketing Board 
Sector Private
Country United Kingdom
Start 10/2018 
End 09/2022
 
Description Further development of infectious bronchitis virus vaccines adapted for production in cell culture
Amount £262,701 (GBP)
Funding ID BB/R019576/1 
Organisation Biotechnology and Biological Sciences Research Council (BBSRC) 
Sector Public
Country United Kingdom
Start 10/2018 
End 03/2020
 
Description Taiwan Partnering Award
Amount £40,700 (GBP)
Funding ID BB/S020624/1 
Organisation Biotechnology and Biological Sciences Research Council (BBSRC) 
Sector Public
Country United Kingdom
Start  
 
Title Optimisation of chicken kidney cell culture preparation 
Description We have optimised the method of extracting kidneys and preparing chicken kidney (CK) cell cultures from chickens. CK cells are used for in vitro assays to study avian viruses, in particular infectious bronchitis virus. 
Type Of Material Cell line 
Year Produced 2017 
Provided To Others? No  
Impact This has increased the yield of viable cells and the quality of the cell cultures. We can now obtain the number of cells we require using fewer chicken kidneys, therefore requiring fewer chickens to be culled. This is an important development for the 3Rs. We hope to publish this method for others' information. 
 
Title Optimisation of preparation of TOCs from adult chickens 
Description Tracheal organ cultures (TOCs) are prepared from chicken embryos and used to assess growth of avian viruses that are not able to be propagated in cell culture. We have optimised the method of TOCs preparation from 3-week old chickens and are using these to assess viral genetic stability and phenotype in organ cultures that are a similar environment to the site of replication in whole chickens. 
Type Of Material Cell line 
Year Produced 2018 
Provided To Others? No  
Impact We hope that this technique will reduce the number of chickens required for in vivo experiments, in line with the 3Rs, and plan to publish the method in due course. 
 
Description Collaboration with University of Cambridge 
Organisation University of Cambridge
Country United Kingdom 
Sector Academic/University 
PI Contribution We have performed in vitro experiments with infectious bronchitis virus, contributed to data analysis, preparation of manuscripts and discussed further funding opportunities.
Collaborator Contribution Our partners have analysed viral transcription and translation, and host gene expression after infection of host cells with infectious bronchitis virus, prepared manuscripts and discussed further funding opportunities.
Impact Dinan, A., Keep, S., Bickerton, E., Britton, P., Firth, A. and Brierley, I. (2019) Comparative analysis of gene expression in virulent and attenuated strains of infectious bronchitis virus at sub-codon resolution. J. Virol. 93(18) e00714-19.
Start Year 2012
 
Description Identification of conserved B-cell epitopes of highly pathogenic coronaviruses for broadspectrum immunotherapy and vaccine design. 
Organisation Imperial College London
Country United Kingdom 
Sector Academic/University 
PI Contribution We have performed experiments to generate preliminary data for a collaborative grant application, contributed to the writing of the grant proposal and responded to reviewers comments.
Collaborator Contribution My collaborators have performed experiments to generate preliminary data for a collaborative grant application, contributed to the writing of the grant proposal and responded to reviewers comments.
Impact Submission of proposal entitled "Identification of conserved B-cell epitopes of highly pathogenic coronaviruses for broadspectrum immunotherapy and vaccine design" to the "One Health Approaches to Accelerate Vaccine Development" call that forms part of the UK government's commitment to Official Development Assistance (ODA) in October 2017.
Start Year 2017
 
Description Identification of conserved B-cell epitopes of highly pathogenic coronaviruses for broadspectrum immunotherapy and vaccine design. 
Organisation University of Kent
Country United Kingdom 
Sector Academic/University 
PI Contribution We have performed experiments to generate preliminary data for a collaborative grant application, contributed to the writing of the grant proposal and responded to reviewers comments.
Collaborator Contribution My collaborators have performed experiments to generate preliminary data for a collaborative grant application, contributed to the writing of the grant proposal and responded to reviewers comments.
Impact Submission of proposal entitled "Identification of conserved B-cell epitopes of highly pathogenic coronaviruses for broadspectrum immunotherapy and vaccine design" to the "One Health Approaches to Accelerate Vaccine Development" call that forms part of the UK government's commitment to Official Development Assistance (ODA) in October 2017.
Start Year 2017
 
Description Partnering with National Taiwan University 
Organisation National Taiwan University
Country Taiwan, Province of China 
Sector Academic/University 
PI Contribution We have prepared and submitted an application for a BBSRC-Taiwan Partnering Award to study "Assembly of Recombinant Infectious Bronchitis Virus and Determination of Antigenic Sites that Confer Hemagglutination Activity".
Collaborator Contribution The partners conceived the project and got in contact about submitting an application for funding together.
Impact An application has been made for a BBSRC-Taiwan Partnering Award in November 2018.
Start Year 2018
 
Description Careers fair 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Schools
Results and Impact Students from several schools in the region attended the careers fair where we had a stall. Several students requested information about apprenticeships and were interested in possible careers in science.
Year(s) Of Engagement Activity 2011,2012,2013,2014,2015,2016,2017
 
Description Centre of excellence for research on avian diseases (CERAD) 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Other audiences
Results and Impact I was invited to present my work at the UK-China Centre of excellence for research on avian diseases (CERAD) meeting attended by researchers from UK, China and Thailand. There was lots of discussion about the research and future directions, including potential collaborations.
Year(s) Of Engagement Activity 2017
 
Description International Avian Respiratory Disease Conference 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Industry/Business
Results and Impact Michael presented his research at the 2018 International Avian Respiratory Disease Conference in the US to an audience of avian researchers and representatives form the poultry veterinary vaccine industry. This sparked discussion and potential future collaborations
Year(s) Of Engagement Activity 2018
 
Description International Day of Women and Girls in Science 
Form Of Engagement Activity Engagement focused website, blog or social media channel
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Public/other audiences
Results and Impact I gave a quote as a STEM ambassador about why I think it is important to make sure women and girls have the same opportunities in STEM. This was shared on social media to celebrate International Day of Women and Girls in Science, a UN-led initiative.
Year(s) Of Engagement Activity 2019
 
Description Interview for national newspaper 
Form Of Engagement Activity A press release, press conference or response to a media enquiry/interview
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Media (as a channel to the public)
Results and Impact Interview with a journalist from The Telegraph about coronavirus replication and transmission.
Year(s) Of Engagement Activity 2020
 
Description Interview with Associated Press 
Form Of Engagement Activity A press release, press conference or response to a media enquiry/interview
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Media (as a channel to the public)
Results and Impact Interview with a journalist from Associated Press about coronavirus research at The Pirbright Institute.
Year(s) Of Engagement Activity 2020
 
Description Interview with BBC Radio Surrey 
Form Of Engagement Activity A press release, press conference or response to a media enquiry/interview
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Media (as a channel to the public)
Results and Impact Interview with journalists from BBC Radio Surrey on the breakfast show about coronavirus research at The Pirbright Institute.
Year(s) Of Engagement Activity 2020
 
Description Interview with BuzzFeed News 
Form Of Engagement Activity A press release, press conference or response to a media enquiry/interview
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Media (as a channel to the public)
Results and Impact Interview with a journalist from BuzzFeed News about coronavirus research at The Pirbright Institute.
Year(s) Of Engagement Activity 2020
 
Description Invited seminar speaker 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Other audiences
Results and Impact Erica Bickerton was invited to give a seminar at Imperial College London, which sparked questions and discussion afterwards.
Year(s) Of Engagement Activity 2017
 
Description MSc student visit 
Form Of Engagement Activity Participation in an open day or visit at my research institution
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Postgraduate students
Results and Impact I hosted several MSc students from University of Surrey, discussed avian research with them and demonstrated some laboratory techniques. The students gained an understanding of academic research and laboratory work.
Year(s) Of Engagement Activity 2018,2019
 
Description Microbiology Society Annual conference (UK) 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Industry/Business
Results and Impact Poster or oral presentation at Microbiology Society annual conference, a major international event. The participants included researchers from many different areas of science. This provides a platform to promote research to a wider audience as well as discuss the key findings with experts in my own field.
Year(s) Of Engagement Activity 2010,2011,2012,2013,2014,2015,2016,2017,2018,2019
 
Description Microbiology Society Avian Focus Meeting (UK) 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Industry/Business
Results and Impact I gave a 15 minute oral presentation to an audience of around 80 people in 2016 and 2018. The audience consisted of researchers from many different scientific institutions from different areas of avian research. This was a great opportunity to present to experts in this field and gain novel insights into the project and the possible applications of my work.
Year(s) Of Engagement Activity 2016,2018
 
Description Nidovirus symposium 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Other audiences
Results and Impact I presented my work at the Nidovirus Symposium, which is held every three years and brings together researchers from industry and academia from many different countries. I had some interesting discussions with other researchers and built my network.
Year(s) Of Engagement Activity 2017
 
Description Positive Strand Viruses Conference 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Industry/Business
Results and Impact This was a poster presentation. There were several different poster sessions at the conference allowing a large selection of people to view each poster. The audience included world leaders in my research field and important names from groups all over the world. This gave me the opportunity to discuss my work with other scientists on a one to one basis, leading to generation of new ideas for future experiments and collaborations.
Year(s) Of Engagement Activity 2010,2013,2016,2019
 
Description Presentation at Global Alliance for Research on Avian Diseases Conference 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Other audiences
Results and Impact Presentation of my research at the Global Alliance for Research on Avian Diseases Conference in Vietnam to approximately 80 conference participants resulted in discussions about future research directions.
Year(s) Of Engagement Activity 2018
 
Description Presentation at RIVR meetings 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Other audiences
Results and Impact Presentation of my research at Recently Independent Virology Researchers (RIVR) meetings in 2016, 2017, 2018, 2019 and 2020. I discussed my work and future collaborations with other virology researchers working in the UK.
Year(s) Of Engagement Activity 2016,2017,2018,2019,2020
 
Description Presentation to BBSRC Executive Board 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Other audiences
Results and Impact I presented our research to the BBSRC Executive Board when they visited The Pirbright Institute in September 2018 and discussed the impacts of our work..
Year(s) Of Engagement Activity 2018
 
Description Reverse genetics seminar (University of Surrey) 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Postgraduate students
Results and Impact A seminar was given based upon the reverse genetics system of infectious bronchitis virus. The audience was a group of 10 -15 MSc students from University of Surrey. The students were engaged, asked questions about the research and careers in science. We were asked to repeat the seminar for the next year's intake of students.
Year(s) Of Engagement Activity 2013,2014,2015,2016,2017,2018
 
Description School visit (Compton, West Berkshire) 
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 The school visits included working with small groups of a-level biology students demonstrating a technique in the curriculum. They were encouraged to ask us questions about our work and how we use the technique as well as trying it out for themselves.
Year(s) Of Engagement Activity 2012,2013,2014,2015
 
Description Teentech 
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 The annual Teentech student science event in Guildford aims to encourage teenagers to consider science subjects and science careers. Our stand had considerable interest from the schoolchildren who were very enthusiastic. The event stimulated an increased interest in science and research.
Year(s) Of Engagement Activity 2016,2019
 
Description organised workshops at Microbiology Society annual conferences 2014 - 2018 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Other audiences
Results and Impact I co-organised virology division workshops on viral evolution and diversity and RNA viruses with colleagues from the virology division. The workshops were held at the annual Microbiology Society conferences in spring each year. Organisation involved selecting abstracts for oral or poster presentation, ordering the presentations and chairing the sessions.
Year(s) Of Engagement Activity 2015,2016,2017,2018