A One Health approach to pan-valent morbillivirus vaccines

Lead Research Organisation: University of Glasgow
Department Name: MRC Centre for Virus Research


Paramyxoviruses are a family of viruses that pose a significant threat to humans and animals alike. For example, Hendra virus outbreaks in Australia have targeted both racehorses and their trainers, while Nipah virus outbreaks in Malaysia spread through pig farms, resulting in 105 human fatalities and the culling of approximately 1.1 million pigs. While such cross-species transmission events may occur relatively infrequently, when they do occur, the consequences can be devastating for humans and animals alike. The Paramyxovirus family also includes the morbillivirus genus of viruses, possibly the most notable of which are measles virus in humans and rinderpest virus in cattle. Morbilliviruses of animals have a very high potential for cross-species transmission, or "zoonosis". Although originally described as a pathogen of dogs, canine distemper virus (CDV) has jumped species into critically endangered lion, tiger, seal and giant panda populations. A closely related morbillivirus of sheep and goats, peste des petits ruminants virus (PPRV) has caused severe disease outbreaks in saiga antelope in Mongolia, buffalo in India and camels in Ethiopia and Sudan. If we are to combat the future spread of such zoonotic morbilliviruses, we need to develop a better understanding of the process of cross-species transmission and how this can be prevented by vaccination.

Vaccination against one species of morbillivirus can provide immunity to infection with morbilliviruses from other species; a rinderpest vaccine can protect cattle from rinderpest, a goat from PPRV and a dog from CDV. Vaccination triggers the production of neutralising antibodies (nAbs) targeting the surface glycoproteins of the virus, hence, some of these antibodies must recognize determinants that are conserved across diverse morbilliviruses. In this project, we set out to identify the determinants on the viral surface glycoproteins that are recognised by neutralising antibodies. If these binding sites can be identified, we will be able to design better pan-morbilliviral vaccines, vaccines that will protect against cross-species infection with emerging morbilliviruses.

To achieve this goal, we have designed in vitro systems with which we can recreate the cellular environment of the susceptible host species. Morbilliviruses attach to target cells by binding to either of two protein molecules expressed on the cell surface, CD150 or nectin-4. Variations in the amino acid sequences of these molecules are key determinants of which species the viruses target, hence CDV is able to bind to CD150 from many species and thus has a very broad species tropism. Neither CDV nor PPRV bind human CD150 well and hence infect human cells very inefficiently. However, with a small adaptation to the amino acid sequence of the viral glycoproteins, binding is enhanced greatly and the viruses display an enhanced ability to infect human cells. We have developed assays with which we can study the propensity for morbilliviruses to develop such mutations. Moreover, we can assess how efficient antibodies generated in the host by vaccination are at preventing infection with such viruses. In effect, we have developed systems with which we can determine the likelihood of zoonotic transmission occurring and how to prevent it by vaccination.

This project will guide the design of the next generation of pan-morbilliviral vaccines and will develop assays systems with which their likely efficacy in preventing zoonoses can be assessed in vitro. By defining the viral targets for neutralising antibodies and by assessing their function against potentially zoonotic viruses, this project offers a unique opportunity to protect both humans and animals alike against the emerging threat posed by novel morbilliviruses.

Technical Summary

Paramyxoviruses pose a significant emerging threat to humans and animals alike, exemplified by the devastating consequences of Hendra and Nipah virus outbreaks in Australia and Malaysia respectively. The very high potential for zoonotic transmission is further illustrated by the morbillivirus canine distemper virus (CDV). Ostensibly a virus of dogs, CDV has jumped species into critically endangered lion, tiger, seal and giant panda populations. Further, CDV can also infect macaques, suggesting that barriers to zoonotic transmission to humans may be fragile.

Vaccination against one species of morbillivirus can provide immunity to infection with other species, hence a rinderpest vaccine can protect cattle from rinderpest (RPV), a goat from peste des petits ruminants (PPRV) and a dog from CDV, while measles vaccination provides partial protection of non-human primates from infection with CDV. We have found that vaccinated and convalescent sera from animals and humans alike, often contain antibodies that cross-neutralise both homologous and heterologous species of virus and hence must recognise determinants that are conserved across diverse morbilliviruses. In this project, we will identify the epitopes recognised by such antibodies, informing the design of the next generation of pan-morbilliviral vaccines, vaccines that will protect against cross-species infection with emerging morbilliviruses.

To achieve this aim, we have developed in vitro systems with which we can assay neutralising activity against primary morbilliviral strains using target cells bearing both cognate and heterologous viral receptors. Moreover, we have developed in vitro assays with which we can investigate the likelihood of potentially zoonotic viruses emerging, characterise the mutations conferring enhanced transmission and assess the sensitivity of the viruses to cross-neutralising antibodies.

Planned Impact

Informing the design of the next generation of morbilliviral vaccines: Live attenuated morbilliviral vaccines have proven efficacy in preventing, controlling and eradicating morbilliviral diseases. Some of these vaccines induce potent cross-neutralising antibodies. The significance of these observations cannot be overstated, if we can identify the molecular determinants targeted by these antibodies, we can engineer the next generation of "pan-morbilliviral" vaccines that will prevent the emergence of novel morbilliviruses. Researchers may choose to pursue diverse vaccine platforms, from classical live attenuated viruses, to engineered chimp adenoviruses bearing surface glycoproteins or adjuvanted recombinant proteins. Irrespective of the platform technology, the data generated in this project will facilitate enhancement and optimisation of antigenicity, inducing a more potent and cross-protective immune response.

Platform technologies for vaccine efficacy testing: This project will deliver high-throughput quantitative assays that facilitate a rapid assessment of neutralising activity in sera from vaccinates and convalescent individuals against biologically relevant field strains of virus. Moreover, we will deliver target cells that recreate the cellular environment of the host species for which efficacy is being assessed. The novel approach we present to predicting likely emergent strains and evaluating their sensitivity to cross-protective neutralising antibodies presents a unique opportunity for vaccine researchers and manufacturers to optimise vaccine formulations to protect animals and humans against not only "known" pathogens, but also against likely "emerging" pathogens, a novel approach to contingency planning in vaccine development.

Tools, resources and reagents for application in the field: As we build up a map of cross neutralising epitopes on the morbilliviral haemagglutinin proteins, we will develop an antigenic map of the major morbillivirus neutralising epitopes targeted by field sera. Hence, when novel field strains of virus emerge that appear to evade immunity induced by existing vaccines, it should be possible to predict which epitopes confer resistance upon the escape mutants. By building structure predictions based on the solved structures of measles virus haemagglutinin, it will be possible to make informed decisions regarding future vaccine compositions. Ultimately, it should be possible to generate a database of morbillivirus mutations that can be used for epidemiological monitoring of inter-species and zoonotic morbillivirus emergence.

Advancement of scientific knowledge within both the academic community and general public: The academic community will benefit directly from this research through an advancement of our understanding of morbillivirus pathogenesis and immunity, the nature of the virus-receptor interaction and the mechanisms of virus neutralisation, stimulating the emergence of new areas of research. At present, the field of paramyxovirus research is relatively small in the UK. This project will build capacity for animal and human morbilliviral research, establishing a strong collaboration between two of the UK's major centres for virology, the Centre for Virus Research and The Pirbright Institute.

Development and showcasing of novel technologies and assay platforms: We have developed a number of novel strategies for integrating high throughout fusion and budding assays with immune-surveillance (particularly neutralising antibodies) and the prediction of zoonotic transmission events. By publishing and presenting these approaches we will foster their application in academia, industry and one-health care. Our close links with clinicians in human and animal health will enable us to expand the outlook of researchers in both human and animal diseases, providing an informed perspective on zoonoses, their animal origins and how to prevent them happening.


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Description In initial studies, we have demonstrated that a morbillivirus that causes disease in small ruminants such as sheep and goats, known as peste des petits ruminants or "PPRV" (a virus that is related to measles virus - the human morbillivirus), could, through minor changes in a particular protein, overcome barriers that currently prevent it from entering human cells.
In collaboration with Dr Dalan Bailey from the Pirbright Institute, the University of Cambridge and other institutions, this study identified that a small change to the PPRV surface protein enables it to use the human receptor to gain entry into human cells.
Using our existing understanding of how these proteins interacted, and previous sequencing and structural studies, the study was able to identify and confirm that a single amino acid in the PPRV hemagglutinin enabled infection of human cells. While this does not mean the virus would have the potential to cause disease in humans as there are many other factors required for the virus to successfully replicate and cause clinical symptoms, it does indicate that these viruses have zoonotic potential given the right mutations and conditions.
The laboratory techniques used to identify these changes were novel. Significantly, the approach used negated the requirement for live infectious virus and high containment laboratories and meant modified live virus with potential zoonotic capability were not, and will not, be generated.
The research was published in the Journal of Virology.
The successful eradication of rinderpest virus (a large ruminant morbillivirus) and the potential eradication of measles has led to concerns that small changes in other morbilliviruses, including PPRV, could enable the disease to emerge in so-called 'vacated ecological niches'. Improving understanding of the determinants of host-range, particularly at the genetic and structural level, will allow researchers to monitor the emergence of these viruses with increased accuracy.
Exploitation Route All the techniques used in the study are described fully in the published version, including comprehensive supporting online materials. The reagents used are freely available.
Sectors Agriculture, Food and Drink,Healthcare,Pharmaceuticals and Medical Biotechnology

URL https://www.pirbright.ac.uk/news/2018/10/pirbright-scientists-use-ground-breaking-techniques-identify-how-ruminant-virus-can
Description Early findings published in Journal of Virology in collaboration with Dr Dalan Bailey, The Pirbright Institute, we discussed in the the podcast This Week in Virology (TWIV). WIV appeals to a broad audience with both academic and non-academic backgrounds, including the wider scientifically literate general public. During the podcast episode (TWIV Episode 532: Morbillivirus had a little lamb - http://www.microbe.tv/twiv/twiv-532/), the hosts discuss the impact of our studies and the opportunities for using such approaches to gain-of-function analyses that do not involve the generation of potentially hazardous in vitro adapted pathogens.
First Year Of Impact 2019
Sector Agriculture, Food and Drink,Healthcare
Impact Types Cultural

Description European Medicines Agency Ad Hoc Advisory Group
Geographic Reach Europe 
Policy Influence Type Participation in a advisory committee
Impact The advisory committee was tasked with assessing the risks associated with the presence of contaminating retroviruses in feline and canine vaccines. The risks were assessed and guidelines for the Animal Healthcare Industry, the European Medicines Agency, veterinary practitioners and the public were drafted and published. The committee engaged in productive dialogue with representatives of the Animal Healthcare Industry, ensuring that vaccine products designed for use in cats and dogs met the highest possible standards of safety.
URL https://www.ema.europa.eu/documents/regulatory-procedural-guideline/cvmp-risk-management-strategy-ma...
Title Development of viral pseudotypes bearing morbilliviral glycoproteins 
Description We are developing and optimising assay systems with which morbilliviral glycoproteins H (hemagglutinin) and F (fusion) can be expressed on the surface of vesicular stomatitis virus (VSV) particles. The particles carry a luciferase marker gene and hence may be used to measure virus neutralising antibodies with high sensitivity and under reduced (CL2) containment. 
Type Of Material Technology assay or reagent 
Year Produced 2016 
Provided To Others? Yes  
Impact The techniques have been used to investigate the seroprevalence of morbilliviruses in livestock in Tanzania, revealing widespread infection of cattle, sheep and goats with diverse morbilliviruses. The techniques have also been used to detect low levels of neutralising antibodies in peste des petits ruminants virus (PPRV)-vaccinated cattle prior to challenge with rinderpest virus (RPV), and to measure cross-neutralisation of measles virus (MeV) by sera from goats exposed to PPRV. We have exported the technique to other laboratories in London and Thailand. 
Description Exchange of novel hemagglutinin and fusion protein expression constructs with Pirbright 
Organisation The Pirbright Institute
Country United Kingdom 
Sector Academic/University 
PI Contribution We have prepared expression vectors bearing clones of the hemagglutinin and fusion protein genes from the four major lineages of peste des petits ruminants (PPRV) circulating globally. Similarly, we prepared H and F expression constructs from diverse global lineages of canine distemper virus (CDV), and from the virulent RBOK strain of rinderpest virus (RPV). These constructs were shared with Dr. Dalan Bailey, our collaborator at the Pirbright Institute.
Collaborator Contribution Dr. Dalan Bailey, our collaborator at the Pirbright Institute, will use the PPRV, CDV and RPV clones to investigate the virus-receptor interaction, the determinants that govern the emergence of potential zoonotic threats and the rol of escape from neutralising antibodies in driving the emergence of novel viral variants.
Impact Initial findings underpinned by this collaboration were published in Journal of Virology - https://jvi.asm.org/content/92/23/e01248-18
Start Year 2017
Description Provision of sera and RNAs for analysis of PPRV lineage-specific neutralisation 
Organisation FAO/IAEA Agriculture and Biotechnology Laboratories
Country Austria 
Sector Private 
PI Contribution We have conducted assays for neutralising antibodies using sera from PPRV vaccinated and infected animals. We have cloned and sequenced H and F genes from isolates of PRPV from distinct lineages.
Collaborator Contribution Researchers in the IAEA laboratories have provided sera and RNAs to facilitate our studies on lineage specific neutralisation of PPRV.
Impact Manuscript describing lineage specific neutralisation in preparation.
Start Year 2013
Description Screening of ruminant sera from Tanzania for morbillivirus-specific neutralising antibodies 
Organisation University of Glasgow
Department Institute of Biodiversity, Animal Health and Comparative Medicine
Country United Kingdom 
Sector Multiple 
PI Contribution We have screened sera from diverse wildlife species and domesticated livestock for morbillivirus-specific antibodies.
Collaborator Contribution Our colleagues in IBAHCM (led by Prof. Sarah Cleaveland) provide access to sera from historical and ongoing studies on a range of pathogens for morbillivirus antibody screening. They have advised on economic and cultural factors influencing the likelihood of exposure to pathogens in pastoralist communities in Tanzania. The collaboration has extended to include Dr Harriet Auty (Scotland's Rural College); Dr. Furaha Mramba, Tanzania Veterinary Laboratory; Prof. Rudovick Kazwala, Sokoine University of Agriculture; Dr. Julius Keyyu and Dr. Robert Fyumagwa, Tanzania Wildlife Research Institute. Dr. Harriet Auty is a veterinary epidemiologist investigating the dynamics, impacts and control of vector-borne diseases and zoonotic diseases of livestock. She is a research fellow with EPIC - the Scottish Government's Centre of Expertise on Animal Disease Outbreaks. Dr. Auty will lead field studies in the coming years. Prof. Sarah Cleaveland is an internationally renowned expert on rabies and zoonotic diseases. Elected a fellow of the Royal Society in 2016, she has worked extensively on the control of neglected tropical diseases in Africa, building and sustaining partnerships with African institutions that will create positive changes in the lives of disadvantaged people. Dr. Furaha Mramba, head of the Tanzania Veterinary Laboratory Agency, liaises with the Ministry of Agriculture, Livestock and Fisheries in applying results to the development of control strategies in Tanzania, and supports coordination of field work in Tanzania.
Impact This collaboration provided some of the original data that informed the development of the the pseudotype based neutralisation assay. Shared master's students have been supervised and a PhD student has been appointed under a separate funding programme. Data generated through this collaboration underpinned the successful application for follow-on funding from the BBSRC and a new sample set collected in 2016 is integral to the first year of the new study.
Start Year 2013
Description Screening of sera from PPRV and RPV exposed/vaccinated ruminants for neutralising antibodies 
Organisation The Pirbright Institute
Country United Kingdom 
Sector Academic/University 
PI Contribution Our laboratory conducts the screening of sera for neutralising antibodies against morbilliviruses. We have been able to generate data for their recent vaccine studies using our enhanced sensitivity assay. The data from this collaboration are due to be published in Journal of Virology in the near future.
Collaborator Contribution The Pirbirght Institute have provided serum samples for analysis, diagnostic standards and valuable advice on assay development.
Impact Manuscript submitted and in review: "Protection of cattle against rinderpest by vaccination with wild type but not attenuated strains of peste des petits ruminants virus". Dr. Michael D. Baron, Barbara Holzer , Sophia Hodgson (Pirbright Institute) & Nicola Logan, Brian J. Willett (University of Glasgow).
Start Year 2014
Description Screening of wildlife sera for antibodies against canine distemper virus (CDV) 
Organisation Wildlife Conservation Society
Country United States 
Sector Charity/Non Profit 
PI Contribution We have conducted screening of wildlife sera in Russia for exposure to CDV, assessing the potential threat of zoonotic spread to susceptible hosts (such as endangered tigers). We have prepared viral pseudotypes using genes derived from field strains of CDV that are circulating in Russia, enabling us to measure neutralising antibodies against biologically relevant strains of virus.
Collaborator Contribution Our partners in the WCS have provided serum samples for serological testing and host nucleic acid samples for amplification of viral sequences to generate viral pseudotypes bearing envelope glycoproteins from circulating visual strains in Russia.
Impact We have validated out pseudo typing strategy using sera provided by WCS researchers, generating antibody titre data in return. We have generated novel viral sequences for phylogenetic studies, complementing similar studies conducted by the WCS.
Start Year 2014
Description Laboratory techniques training session 
Form Of Engagement Activity Participation in an activity, workshop or similar
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Professional Practitioners
Results and Impact Visit to the Nelson Mandela African Institute of Science and Technology in Tanzania to host laboratory workshop during which local scientists learned ELISA techniques used to measure Rift Valley Fever Virus antibodies in sera.
Year(s) Of Engagement Activity 2013
Description Postgraduate lectures 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Postgraduate students
Results and Impact Delivery of postgraduate teaching on taught component of masters degree and laboratory projects on morbilliviruses.
Year(s) Of Engagement Activity 2015,2016
Description Social media acitvities 
Form Of Engagement Activity Engagement focused website, blog or social media channel
Part Of Official Scheme? No
Geographic Reach International
Primary Audience Media (as a channel to the public)
Results and Impact Whenever significant progress is made in the research project, our findings are publicised through CVR and Pirbright social media outlets, and through our laboratory twitter account @Animal_Viruses. We direct followers to the source publication, to News articles covering the publication, and to comment articles. For example, our recent publication in Journal of Virology was discussed on the TWIV podcast (This Week In Virology).
Year(s) Of Engagement Activity 2018,2019
Description Undergraduate lectures 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach National
Primary Audience Undergraduate students
Results and Impact The provision of research-led undergraduate lectures on morbilliviruses.
Year(s) Of Engagement Activity 2015,2016