Glycoengineering of Veterinary Vaccines

Lead Research Organisation: London Sch of Hygiene and Trop Medicine
Department Name: Infectious and Tropical Diseases


A healthily maintained livestock is essential for the economy and prosperity of the UK. Additionally some infected livestock are the source of human diseases, particularly through foodborne infections. Historically, vaccines have been the most successful and effective intervention to reduce the burden of infectious diseases in humans. By contrast, the application of vaccines in veterinary medicine is rudimentary, mainly due to the economic necessity for reduced costs to vaccinate animals and because our knowledge of the pathogens that cause animal diseases lags behind that of human counterparts.

A defining characteristic of a successful vaccine is the ability to evoke long-lasting protective immunity with minimal side effects. Many of the most successful human vaccines are glycoconjugates, a combination of a protein coupled to a glycan, which induces both a T-cell dependent and independent immune response generating a protective and lasting immunity. Examples of currently licensed human glycoconjugate vaccines include those against Haemophilus influenzae, Neisseria meningitidis and Streptococcus pneumoniae, in which glycans (lipopolysaccharides or capsular polysaccharides) are chemically coupled to immunogenic carrier proteins. However, the production of these vaccines requires multistep procedures that are often complex and expensive, and can exhibit batch-to-batch variation.

We recently developed Protein Glycan Coupling Technology (PGCT) that can overcome the complex procedures required for chemically synthesising glycoconjugate vaccines by expressing the vaccine in an Escherichia coli cell in a single-step procedure. The advantages of applying PGCT to veterinary vaccines are (i) glycoconjugate vaccines can be produced at low cost, (ii) the flexibility of coupling "any glycan" with "any protein" facilitates the production of vaccine combinations providing the opportunity to evaluate a greater variety of vaccine candidates, and (iii) combination vaccines against more than one disease can be produced, further reducing cost and obviating the need to administer multiple vaccines (or antibiotics).

In this study we will use PGCT to produce inexpensive triple combination poultry vaccines to reduce infection from E. coli, Salmonella, Campylobacter jejuni/coli and C. perfringens. This will not only protect poultry flocks from severe disease but would also protect the human population from the most common foodborne infections including those caused by Salmonella and Campylobacter. In addition we will construct and evaluate a dual Coxiella/C. perfringens vaccine to protect cattle, sheep and goats against severe disease. This vaccine would also prevent the spread of Q-fever to humans, which is caused by the highly infectious Coxiella burnetii pathogen. The principles developed in this proposal could subsequently be widely applied to produce inexpensive efficacious vaccines against most animal species and promise to break new ground in veterinary vaccine production.

Technical Summary

An unmet need in veterinary vaccinology is the production of low cost effective vaccines that can protect against multiple infectious agents. We will aim to capitalise on our recent characterisation of a novel N-linked general glycosylation system in Campylobacter jejuni that can be used to engineer multiple combinations of glyco-modified proteins in different bacteria including E. coli and Salmonella species. This protein glycan coupling technology (PGCT) is proven in the production of human vaccines, but has yet to be applied to veterinary vaccines. We propose to use PGCT to construct dual and triple combination vaccines to reduce the carriage of Salmonella, Campylobacter, E. coli and Clostridium perfringens in poultry. The engineered constructs will also be used to investigate basic immunological responses in chickens to these pathogens, and will be tested for protection in chickens against infection.

To expedite the application of PGCT we will develop a more detailed understanding of glycobiosynthetic pathways in pathogenic bacteria including Coxiella burnetii. The novel LPS biosynthetic pathway will be thoroughly characterised by genetic, chemical and structural analyses. We will clone and express the LPS from C. burnetii in E. coli and couple this to genetic toxoids from C. perfringens including deactivated NetB to produce a dual vaccine. We will assess vaccine candidates produced by examining markers for humoral and cellular immunity, and the ability to induce protective immunity against C. perfringens toxins and C. burnetii in mice. In parallel with the development of the stated veterinary vaccines, we will use the opportunity to further innovate PGCT. We aim to improve the utility, efficacy and general applicability of PGCT for glycoconjugate vaccinology and for further glycobiotechnological applications. Additionally, we will foster our industrial collaborations to fully exploit the vaccines and innovations derived from this research.

Planned Impact

The economy
The knowledge generated in the program and application of the research would clearly benefit the poultry and livestock industry as well as farming communities. Ultimately, through reduced occurrence of food poisoning, the knowledge gained in this study will improve the health and wealth of the nation. The reduction of serious infections in livestock coupled with the development and manufacture of novel vaccines will provide significant benefits to the UK economy. The impacts of the research program are potentially enormous and manifold. Vaccines are proven for the control of infectious diseases in both humans and in animals, and suitably designed vaccines will reduce our reliance on antibiotics. With the UK livestock industry (including cows, pigs, sheep, poultry and fish) estimated to have an annual value of over £14bn in 2013, smart design vaccines will have direct benefits for the UK economy. Chickens alone are the world's most popular food animal with global poultry production tripling in the past 20 years and will continue to increase. Therefore, farmers and the agricultural industry will significantly benefit from cheaper more effective vaccines that target livestock.

The general public
The general public will benefit from less food poisoning in the reduction of C. jejuni and C. coli in the food chain, with the resultant economic benefit to the UK economy in terms of improved productivity. C. perfringens is major cause of disease in domesticated livestock ranging from enterotoxaemia in sheep, goals and calves to necrotic enteritis in poultry, a disease which is emerging following the EU ban on the use of antibiotics to promote growth. An effective poultry vaccine would also discourage the indiscriminate use of antimicrobials in livestock and contribute to reducing antimicrobial resistance. Thus a significant impact will be the reduction in antibiotic use, a key government policy and priority Coxiella is a zoonotic agent therefore an effective animal vaccine would reduce transmission of Q fever to humans. Additionally, there would be a market for a human Coxiella vaccine to protect workers likely to come into contact with infected animals and where Q fever is endemic as well as for defense purposes. Therefore, the proposal will considerably enhance the quality of life and improve the economic competitiveness of the UK.

Academic and industrial organisations
The development of PGCT would enhance the commercial private sector for the production of vaccines and potentially for glycoengineering human therapeutics. We have close links with Zoetis, Merck, Glycovaxyn, VaxAlta and Malicisbo and will use licensing agreements through our respective technology transfer offices to ensure pipelines to vaccine production and exploitation are in place. Developing a basic understanding of the glycobiosynthetic pathways for the pathogens in this study will not only be important for understanding pathogenesis and vaccine production, but has other practical applications. The inhibition of bacterial glycosyltransferases is a useful target to disable the pathogenic bacteria providing a novel approach for antimicrobial development termed "glycobiotics". Additionally, bacterial glycans are often surface exposed and specific to individual species or virulent clones providing improved diagnostics benefitting human and veterinary health. The technology developed through may have enormous implications for policy makers to future disease outbreaks and impact on exports.

Training opportunities
The consortium will employ and train and develop a cohort of scientists with diverse experience with a "one health" mentality that can be applied in academia, the public sector and industry. The multidisciplinary team will add to the UK science base in an important and economically vital research area.
Description Several combined glycoconjugate vaccine candidate have been constructed using Protein Glycan Coupling Technology that are currently being tested in chickens Routes for the scalable chemical synthesis of virenose monoaccharide have been devised. Methods for the generation of the anomeric sugar phosphate therefrom are in hand, with extension to putative sugar nucleotide building blocks to begin shortly, in support of studies at Exeter Synthetic virenose disaccharides, conjugated to hepatitis B VLPs, have been immunised into rabbits, with bloods for analysis due April 2018.
First Year Of Impact 2017
Sector Agriculture, Food and Drink,Manufacturing, including Industrial Biotechology
Description Bovine Tuberculosis
Amount £11,051,222 (GBP)
Funding ID BB/N004590/1 
Organisation Biotechnology and Biological Sciences Research Council (BBSRC) 
Sector Public
Country United Kingdom
Start 09/2015 
End 09/2019
Description Developing the E. coli GlycoCell
Amount £455,000 (GBP)
Funding ID BB/R008124/1 
Organisation Biotechnology and Biological Sciences Research Council (BBSRC) 
Sector Public
Country United Kingdom
Start 06/2018 
End 06/2021
Description Universal protection against Streptococcus pnuemoniae
Amount £989,000 (GBP)
Funding ID MR/R001871/1 
Organisation Medical Research Council (MRC) 
Sector Academic/University
Country United Kingdom
Start 04/2018 
End 04/2021
Description Zoetis Development of poultry vaccines 
Organisation Zoetis
Country United States 
Sector Private 
PI Contribution Technology to make cheaper more effective vaccines
Collaborator Contribution Testing of vaccines in poultry by Zoetis
Impact Early stage as vaccines are being prepared
Start Year 2017
Title Bacterial Glycotoolbox 
Description Fine tuning of the E. coli cell to express and produce bacterial glycans 
IP Reference GB1603963.8 
Protection Patent granted
Year Protection Granted 2016
Licensed Commercial In Confidence
Impact The establishment of vaccine technology platform to engineer low cost recombinant glycoconjugate vaccines
Title Whole Cell Vaccines 
Description Development of Protein Glycan Coupling Technology for vaccines for veterinary purposes 
IP Reference GB1603958.8 
Protection Patent application published
Year Protection Granted 2016
Licensed Commercial In Confidence
Impact The establishment of vaccine technology platform to engineer low cost recombinant glycoconjugate vaccines
Company Name ArcVax 
Description A spin out company from LSHTM was set up in Dec 2016 to using Protein Glycan Coupling Technology to produce low cost glycoconjugate vaccines. Currently investment is being sought. 
Year Established 2016 
Impact To rapidly develop candidate glycoconjugate vaccines for the Veterinary Vaccine Market, eg multiprotective vaccines for poultry.
Description Feature for BBC2 Food Detectives programme 
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 Public/other audiences
Results and Impact Professor Stevens took part in a BBC2 Food Detectives programme which featured Campylobacter in chickens. He helped to design a survey of Campylobacter contamination in a sample of fresh chicken obtained from Scottish retailers and interpret the findings. This were close to Food Standards Agency findings, which were broadcast as part of an interview with Prof. Alice Roberts. The interview took place at The Roslin Institute and included a lay description of Campylobacter, the importance of chickens as a reservoir of infection and strategies by which consumers can protect themselves. It also highlighted BBSRC-funded work at Roslin to mitigate the problem based on genetic selection (this project) and vaccines (other BBSRC projects held by Prof. Stevens).
Year(s) Of Engagement Activity 2016
Description Public lecture entitled Confronting the Microbial Menace in Our Food'. Professor Mark Stevens 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? No
Geographic Reach Regional
Primary Audience Schools
Results and Impact The inaugural lecture of Professor Mark Stevens was held on 30 October 2017 on his BBSRC-funded research to identify bacterial and host factors influencing the ability of Salmonella, Campylobacter and E. coli to colonise farm animals and cause disease. It was attended by children from local schools, members of the public, students at The Roslin Institute and wider University of Edinburgh and posted online.
Year(s) Of Engagement Activity 2017,2018