VACCINE: Development of Novel BRSV Pre-Fusion Protein Recombinant Bovine Vaccine.

Lead Research Organisation: Queen's University of Belfast
Department Name: Centre for Experimental Medicine

Abstract

Bovine respiratory syncytial virus (BRSV) is a respiratory virus associated bovine respiratory disease complex, which is a major problem for the cattle industry. It results in considerable morbidity and mortality in cattle, and particularly young calves, worldwide. Efficacy of current vaccines is limited and there is a major need for a much more effective vaccine. BRSV is highly similar to Human RSV (HRSV) which is the most common cause of hospitalization in young infants and causes severe disease in elderly. No HRSV vaccines are available to date due to poor efficacy and safety issues. No HRSV vaccines are available to date due to poor efficacy and safety issues. In cattle the response of the immune system of current vaccines is compromised by the presence of maternal antibodies which neutralise the vaccine. This current proposal will target the exploitation of a combination of novel technologies to generate novel BRSV vaccines. Specifically, we will bring together world-renowned expertise in paramyxovirus reverse genetics, structure-based vaccine antigen design, and vaccine development to generate a new generation of BRSV vaccines that we are confident will circumvent many of the limitations of current vaccines. This will include the use of Sendai virus (SeV, a mouse virus) and parainfluenza virus type 5 (PIV5, a dog virus) as vehicles (vectors) to deliver a BRSV gene into the cell and produce the protein (antigen) from this. As neither of these viruses is found in cattle, they infect bovine cells but do not cause disease in cattle and there is no known cross-reactivity with bovine antibodies, problems associated with the presence of maternal antibodies will be circumvented. The use of SeV and PIV5 will facilitate development of a prime/boost vaccine regimen, which will circumvent a virus vector-specific immune response induced by the 1st vaccine administration, as both viruses are antigenically distinct. Such a regimen is likely to maximize immune responses to the BRSV antigen. We will develop both replication competent and incompetent viruses for both SeV and PIV5. The latter are produced by producing virus infectious particles that have one of the genes for further replication removed. However, these particles can enter cells and produce BRSV proteins. Our cumulative data, both published and unpublished, confirms that both replication competent viruses are independently highly efficient vectors for human RSV vaccine antigens in animal models. The development of replication-incompetent SeV and PIV5 viral vectors will address some safety concerns related to environmental dissemination of recombinant viruses, and will involve the generation of novel rescue technologies (production of virus from copies of the virus genes) for both viruses. BRSV and HRSV contain a protein called the fusion (F) protein which allows the virus to get into cells and is in a different form on the virion surface (pre-fusion) compared to when it interacts with a cell (post-fusion). The F protein induces a response by the immune system and the pre-fusion form has recently been shown to produce a several fold higher immune response than the post fusion form (in an experimental HRSV vaccine. We will exploit this finding to design and generate the most highly immunogenic BRSV vaccine, based on the use of a pre-fusion stable F protein. Precedence for this was demonstrated for human RSV pre-fusion F-based experimental vaccine, which was shown to be many fold superior to native F as a vaccine antigen. Finally, we will bring all of these complementary but disparate elements together to develop novel BRSV vaccines and vaccine regimens and test their efficacy in the final host target for the use of these vaccines, i.e., calves. These vaccines and the results from their study will also serve as the basis for design of effective human vaccines for HRSV.

Technical Summary

Bovine respiratory syncytial virus (BRSV) is associated bovine respiratory disease complex, a major problem for the cattle industry, and is closely related to human RSV (HRSV). It results in considerable morbidity and mortality in calves worldwide. Efficacy of current vaccines is limited because they are compromised by maternal antibodies in calves. This proposal will exploit of a combination of novel technologies to generate new BRSV vaccines. We will bring together world-renowned expertise in paramyxovirus reverse genetics, structure-based vaccine antigen design, and vaccine development to generate a new generation of BRSV vaccines designed to circumvent the limitations of current vaccines. This will include the use of Sendai virus (SeV) and parainfluenza virus type 5 (PIV5) as vectors for BRSV vaccine antigens. As neither of these viruses is found in cattle and there is no known cross-reactivity with bovine antibodies, problems associated with maternal antibodies will be circumvented. Use of SeV and PIV5 will facilitate development of a prime/boost vaccine regimen, which will circumvent virus vector-specific immunity induced by the 1st vaccine administration, as both viruses are antigenically distinct. We will develop both replication competent and incompetent viruses for both SeV and PIV5. Replication-incompetent SeV and PIV5 vectors will address safety concerns related to environmental dissemination of recombinant viruses. We will exploit novel structure-based vaccine design to generate the most highly immunogenic BRSV antigen, based on a BRSV pre-fusion stable F protein. Precedence for this was demonstrated for a HRSV pre-fusion F protein, which was many fold superior to native F as a vaccine antigen. We will bring these complementary but disparate elements together to develop novel BRSV vaccines and vaccine regimens and test their efficacy in calves. These vaccines-natural host systems will also serve as the basis for design of effective HRSV vaccines.

Planned Impact

This proposal focuses on the generation of novel bovine respiratory syncytial virus (BRSV) vaccine candidates. BRSV is a respiratory virus associated bovine respiratory disease complex, which is a major problem for the cattle industry and is closely related to human RSV (HRSV) in terms of virus structure and associated clinical disease. It results in considerable morbidity and mortality in cattle, and particularly young calves, worldwide. Efficacy of current vaccines is limited and there is a major need for a much more effective vaccine. The immunogenicity of current vaccines is compromised by the presence of maternal antibodies. This current proposal will target the exploitation of a combination of novel technologies to generate novel BRSV vaccines. Specifically, we will bring together world-renowned expertise in paramyxovirus reverse genetics, structure-based vaccine antigen design, and vaccine development to generate a new generation of BRSV vaccines that we are confident will circumvent many of the limitations of current vaccines. This will include the use of Sendai virus (SeV) and parainfluenza virus type 5 (PIV5) as vectors to deliver BRSV vaccine antigens. As neither of these viruses is found in cattle and there is no known cross-reactivity with bovine antibodies, problems associated with the presence of maternal antibodies will be circumvented. The use of SeV and PIV5 will facilitate development of a prime/boost vaccine regimen, which will circumvent virus vector-specific immunity induced by the 1st vaccine administration, as both viruses are antigenically distinct. Such a regimen is likely to maximize immune responses to the BRSV antigen. We will develop both replication competent and incompetent viruses for both SeV and PIV5. Our cumulative data, both published and unpublished, confirms that both replication competent viruses are independently highly efficient vectors for human RSV vaccine antigens in animal models. The development of replication-incompetent SeV and PIV5 viral vectors will address some safety concerns related to environmental dissemination of recombinant viruses, and will involve the generation of novel rescue technologies for both viruses. We will exploit novel state-of-the-art structure-based vaccine design to generate the most highly immunogenic BRSV antigen, based on the use of a pre-fusion stable F protein construct. Precedence for this was demonstrated for a human RSV pre-fusion F-based vaccine, which was shown to be many fold superior to native F as a vaccine antigen (Science. 2013 342(6158):592-8; McLellan JS et al,Science. 2013 May 31;340(6136):1113-7). Finally, we will bring all of these complementary but disparate elements together to develop novel BRSV vaccines and vaccine regimens and test their efficacy in the final host target for the use of these vaccines, i.e., calves.
Therefore, reasonable outcomes of this project will include:
- novel rescue technologies for replication incompetent SeV and PIV5 vectors;
- novel BRSV vaccines based on replication competent and incompetent SeV and PIV5 vectors expressing BRSV pre-fusion F;
- a novel vaccine regimen to induce efficient protective immunity against BRSV, even in the presence of maternal antibodies;
- a novel BRSV vaccine and regimen with sufficient data to justify commercial development.
- in view of the considerable similarities between BRSV and HRSV, both virologically and clinically, the data derived from this study will likely inform future vaccine strategies for HRSV.
As such, this project is likely to have a considerable impact on UK industry, both biotech and agricultural, and increase the reputation of the UK as a world leader in vaccine design and technologies that directly impact on society.

Publications

10 25 50
 
Description We have successfully modified our rescue system for generating recombinant Sendai viruses expressing heterologous genes. This will greatly help our ability to exploit this technology platform for the generation of novel vaccines against other infectious diseases.
Exploitation Route The modified rescue system for Sendai virus will help us generate the viruses we require to meet the milestones of the project. It will also greatly help us exploit this virus vector platform for the generation of novel vaccines against other viruses.
Sectors Agriculture, Food and Drink,Healthcare,Pharmaceuticals and Medical Biotechnology

 
Description DAERA PhD studentship scheme 2018
Amount £62,106 (GBP)
Organisation Northern Ireland Department of Agriculture, Environment and Rural Affairs (DAERA) 
Sector Public
Country United Kingdom
Start 09/2018 
End 08/2021
 
Title Bovine RSV calf challenge model 
Description As part of Aim 4 of this project, a major milestone was the development of a bovine RSV challenge model in calves. Our collaborators in AFBI, under the direction of Prof. Cosby, have successfully developed the challenge model, as demonstrated by the induction of strong clinical symptoms and extensive lung pathology in calves following infection with bRSV strain 274, which is a wild type Swedish isolate. The challenge virus was delivered by aerosolisation. Clinical symptoms included nasal mucus secretion, raised rectal temperatures (peak 40oC) and increased respiratory rate, peaking at 6-7 days post infection. Lung pathology was demonstrated by macroscopic lesions and histopathology. qRT-PCR demonstrated robust virus growth kinetics in nasal tissues, as determined by daily sampling, and high virus titres were determined in lung tissues at necropsy and associated with strong disperse immunofluorescence in lung tissues. Importantly, the challenge virus was generated by passage in foetal bovine lung cells without subsequent passage in calves, as proposed in the initial application. Thus, the number of animals used for this part of the project was reduced relative to the anticipated numbers. Furthermore this challenge model was effective even in the presence of moderate levels of maternal antibody. 
Type Of Material Model of mechanisms or symptoms - mammalian in vivo 
Year Produced 2018 
Provided To Others? No  
Impact As this model has only recently been established, there are no impacts to report as yet. 
 
Title Improved rescue system for recombinant Sendai virus 
Description As part of this project, we have systematically revisited our Sendai virus (SeV) reverse genetics protocol and optimized conditions to rescue recombinant Sendai viruses expressing heterologous genes. This included introducing a hammerhead ribozyme immediately upstream of the Sendai virus genome leader sequence in the infectious clone and mutations in the artificial EIS sequence designed to prevent homologous recombination associated with the use of vaccinia virus expressing T7 polymerase (which drives transcrption of the infectious clone and helper plasmids). We have also included the use of LLC-MK2 cells as part of the protocol. Recombinant SeVs expressing eGFP and glycoproteins from other paramyxoviruses have been successfully rescued and we are exploring possibilities for a patent submission on our updated protocol. 
Type Of Material Technology assay or reagent 
Year Produced 2019 
Provided To Others? No  
Impact We have exploited our modified SeV rescue system to generate rSeVs expressing eGFP and a glycoprotein from a different paramyxovirus. 
 
Description AFBI 
Organisation Agri-Food and Biosciences Institute
Country United Kingdom 
Sector Public 
PI Contribution I am the project coordinator and have brought together a team of international experts in vaccine generation and development with a view to developing a novel vaccine against bovine RSV. My team brings the Sendai virus vaccine vector platform to the table.
Collaborator Contribution AFBI provide expertise in calf models of bRSV infection and vaccine immunogenicity and protective efficacy testing.
Impact There are no outputs from this collaboration yet.
Start Year 2017
 
Description University of Georgia 
Organisation University of Georgia
Country United States 
Sector Academic/University 
PI Contribution I am the project coordinator and have brought together a team of international experts in vaccine generation and development with a view to developing a novel vaccine against bovine RSV. My team brings the Sendai virus vaccine vector platform to the table.
Collaborator Contribution My collaborator brings the PIV5 vaccine vector platform to the table for this grant.
Impact There are not outcomes yet from this project.
Start Year 2017
 
Description Vaccine Research Centre, NIAID, NIH, USA 
Organisation National Institutes of Health (NIH)
Department Vaccine Research Center (VRC)
Country United States 
Sector Public 
PI Contribution I am the project coordinator and have brought together a team of international experts in vaccine generation and development with a view to developing a novel vaccine against bovine RSV. My team brings the Sendai virus vaccine vector platform to the table.
Collaborator Contribution Our collaborators provided the information necessary to generate the bovine RSV pre-fusion stabilised F protein construct.
Impact There are no outputs from this project yet.
Start Year 2017
 
Description Final year undergraduate research project 
Form Of Engagement Activity Participation in an open day or visit at my research institution
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Undergraduate students
Results and Impact A 3rd year MSci student selected my undergraduate year 3 research project, as part of her obligations to fulfill requirements for her MSci degree in Biochemistry from Queen's University Belfast. Because of this experience, she decided to undertake her year long research project as part of the requirements to complete her MSci degree.
Year(s) Of Engagement Activity 2017
 
Description Northern Ireland Science Festival 
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 Public/other audiences
Results and Impact My team provided an overview of the research undertaken in my group, with emphasis on viruses, the diseases that they cause and how they are transmitted. This was part of an open day at the Wellcome Wolfson Institute of Experimental Medicine at Queen's University Belfast, which was part the Northern Ireland Science Festival 2018. The audience ranged in age from young children to adults.
Year(s) Of Engagement Activity 2018
 
Description Summer studentship 
Form Of Engagement Activity Participation in an open day or visit at my research institution
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Undergraduate students
Results and Impact A 2nd year undergraduate student undertook an 8 week summer studentship in my laboratory, working on parts of this project. Following this, she successfully applied for funding from DAERA to undertake her PhD in my laboratory, working on aspects this project.
Year(s) Of Engagement Activity 2017
 
Description Wellcome Wolfson Institute for Experimental Medicine open day "Know Your Enemy: Disease-Focussed Research at Queen's University". 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Public/other audiences
Results and Impact Dr. Power gave a talk on vaccines, providing an overview of the history of vaccines,the amazing successes associated with 2 vaccines (smallpox and rinderpest), the immunology behind vaccines, and the concept of risk/benefit in relation to vaccine use.
Year(s) Of Engagement Activity 2019
URL https://www.nisciencefestival.com/event.php?e=141
 
Description Wellcome Wolfson Institute for Experimental Medicine open day "Know Your Enemy: Disease-Focussed Research at Queen's University". 
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 Public/other audiences
Results and Impact My team provided an overview of the research undertaken in my group, with emphasis on viruses, the diseases that they cause and how they are transmitted. Members of my team devised several games including 'Catch the Sneeze' and 'Virus match-up'. Okido (Breathing Together project partner) provided magazines, soft books and supplies for make-and-do activities, such as making inflatable paper bag lungs. This was part of an open day at the Wellcome Wolfson Institute of Experimental Medicine at Queen's University Belfast, which was part the Northern Ireland Science Festival 2019. The audience ranged in age from young children to adults. Feedback for the event was overwhelmingly positive with attendees mentioning our stand as a highlight of the day.
Year(s) Of Engagement Activity 2019
URL https://www.qub.ac.uk/research-centres/wwiem/EducationandTraining/Outreach/NIScienceFestival2019/
 
Description Work experience secondary school student 
Form Of Engagement Activity Participation in an open day or visit at my research institution
Part Of Official Scheme? No
Geographic Reach Local
Primary Audience Schools
Results and Impact In May 2017, my lab hosted a work experience secondary school student from Bloomfield Collegiate School. The student shadowed different activities in the lab and attended our lab meeting to learn about the research we conduct.
Year(s) Of Engagement Activity 2017