Immunological toolbox for livestock
Lead Research Organisation:
THE PIRBRIGHT INSTITUTE
Department Name: UNLISTED
Abstract
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Technical Summary
unavailable
Planned Impact
unavailable
Organisations
- THE PIRBRIGHT INSTITUTE (Lead Research Organisation)
- Rosalind Franklin Institute (Collaboration)
- UNIVERSITY OF EDINBURGH (Collaboration)
- EMBL European Bioinformatics Institute (EMBL - EBI) (Collaboration)
- Technical University of Denmark (Collaboration)
- Research Complex at Harwell (Collaboration)
- Agricultural Research Council (ARC) (Collaboration)
Publications
Bakshy K
(2021)
Development of polymorphic markers in the immune gene complex loci of cattle.
in Journal of dairy science
Steyn A
(2020)
The Characterization of chIFITMs in Avian Coronavirus Infection In Vivo, Ex Vivo and In Vitro.
in Genes
| Description | The previous immunological toolbox award provided the means to translate our genomic enrichment methods to assemble immune gene complex regions from diverse cattle breeds. Although our collaboration with Kenya Wildlife Service was not productive, our contact across regions of Africa and our collaboration with the CTLGH enabled us to receive and process samples from three different countries and 6 different breeds (Sahiwal, N'Dama, Boran, White Fulani, Angoni, Barotse). In total we have sequenced 22 different animals and are currently assembling these regions of the genome. We will then compare the SNPs across these regions to those being studies and tested in GWAS as part of our USDA-NIFA collaboration and BBSRC award BB/M027155/1. We will then have a much clearer idea of where selection pressures have diversified immune functional variation in these diverse breeds where disease is endemic. We then aim to put all this genetic data together to design future experiments in African breeds that will allow us to identify genetic variation that will inform future breeding strategies to improve natural disease resilience. |
| Exploitation Route | Developing tests to predict how genomic and genetic diversity impact on disease outcome. This could lead to improved breeding strategies. |
| Sectors | Agriculture Food and Drink |
| Description | Aviagen Industry Funding |
| Amount | £422,383 (GBP) |
| Organisation | Aviagen Group |
| Sector | Private |
| Country | United States |
| Start | 08/2019 |
| End | 12/2020 |
| Description | The Pirbright Livestock Antibody Hub |
| Amount | $5,530,900 (USD) |
| Funding ID | INV-003007 |
| Organisation | Bill and Melinda Gates Foundation |
| Sector | Charity/Non Profit |
| Country | United States |
| Start | 09/2019 |
| End | 09/2024 |
| Title | 8 color flow cytometry panel for cattle B cell phenotyping |
| Description | Flow cytometry panel to characterize different B cell subsets in cattle. |
| Type Of Material | Technology assay or reagent |
| Year Produced | 2022 |
| Provided To Others? | Yes |
| Impact | Following the publication of this method, we have been contacted to provide reagents and expertise for cattle B cell phenotyping in the context of diseases, and to host a researcher form a LMIC for training in flow cytometry. |
| Title | Methods to sequence cattle antibodies at different throughputs |
| Description | A suite of methods to isolate cattle B cells, amplify and sequence cattle antibody heavy and light chains, and to analyse antibody repertoires. Workflow has been adapted for low (Sanger), medium (NGS), and high (10X genomics platform) throughput. |
| Type Of Material | Technology assay or reagent |
| Year Produced | 2023 |
| Provided To Others? | Yes |
| Impact | Following the publication of these methods, we have been contacted by several research groups (University of Sao Paulo, Glasgow, ILRI institute) for collaborations on antibody repertoire analysis. |
| URL | https://doi.org/10.3390/vaccines11061099 |
| Title | Recombinant antibody generation pipeline - WM |
| Description | Validated a sequencing protocol at Pirbright for mouse hybridomas, cattle heterohybridomas, and porcine B cells. Vector constructed for humans, mouse, cattle, pigs and chicken to allow ligation of commercially generated antibody gene blocks or PCR products generated by degenerate or 5' RACE PCR. These methods allow the following: • Sequencing mAbs • Making Fab fragments from mAbs • Class switching mAbs • Species switching mAbs |
| Type Of Material | Technology assay or reagent |
| Year Produced | 2018 |
| Provided To Others? | Yes |
| Impact | The methods developed have enabled a standardised pipeline that allows cloning of PCR products or synthetic gene constructs for recombinant antibody generation from various species. This uniform approach avoids development of bespoke protocols for each species thus saving time and resources. |
| URL | http://www.immunologicaltoolbox.co.uk |
| Title | Recombinant antibody generation pipeline - WM |
| Description | • Validated a sequencing protocol at Pirbright for mouse hybridomas, cattle heterohybridomas, and porcine B cells. • Vector constructed for mouse, cattle, pigs and chicken to allow ligation of commercially generated antibody gene blocks or PCR products generated by degenerate or 5' RACE PCR. These methods allows the follows • Sequencing mAbs • Making Fab fragments from mAbs • Class switching mAbs • Species switching mAbs |
| Type Of Material | Antibody |
| Year Produced | 2018 |
| Provided To Others? | Yes |
| Impact | The methods developed have enabled a standardised pipeline that allows cloning of PCR products or synthetic gene constructs for recombinant antibody generation from various species. This uniform approach avoids development of bespoke protocols for each species thus saving time and resources. |
| Title | Recombinant antibody generation pipeline - WM |
| Description | • Validated a sequencing protocol at Pirbright for mouse hybridomas, cattle heterohybridomas, and porcine B cells. • Vector constructed for mouse, cattle, pigs and chicken to allow ligation of commercially generated antibody gene blocks or PCR products generated by degenerate or 5' RACE PCR. These methods allows the follows • Sequencing mAbs • Making Fab fragments from mAbs • Class switching mAbs • Species switching mAbs |
| Type Of Material | Technology assay or reagent |
| Year Produced | 2018 |
| Provided To Others? | Yes |
| Impact | The methods developed have enabled a standardised pipeline that allows cloning of PCR products or synthetic gene constructs for recombinant antibody generation from various species. This uniform approach avoids development of bespoke protocols for each species thus saving time and resources. |
| Title | Full length cattle MHC genes |
| Description | Using our method for full length MHC gene amplification, the largest reference set ever produced from common haplotypes has been sequenced and deposited in the publicly available data IPD-MHC with associated research tools. |
| Type Of Material | Database/Collection of data |
| Year Produced | 2020 |
| Provided To Others? | No |
| Impact | None yet, but it will allow far greater resolution when analysing MHC polymorphism and evolution in cattle populations. |
| URL | https://www.ebi.ac.uk/ipd/mhc/group/BoLA/ |
| Title | Immunological Toolbox website - WM |
| Description | Website repository for important reagents essential in immunology of livestock |
| Type Of Material | Database/Collection of data |
| Year Produced | 2019 |
| Provided To Others? | Yes |
| Impact | The reagents data repository has proved very useful for new starters in our group in enabling quick gathering of information regarding available antibodies and their applications. |
| Title | UK Immunological Toolbox website - WM |
| Description | Website repository of important reagents for veterinary immunology and vaccinology research. |
| Type Of Material | Database/Collection of data |
| Year Produced | 2019 |
| Provided To Others? | Yes |
| Impact | The reagents data repository has proved very useful for new starters in our group in enabling quick gathering of information regarding available antibodies and their applications. The website has proved useful to other researchers internationally as evidenced by requests currently being received from other countries for reagents that are not commercially available. |
| URL | https://www.immunologicaltoolbox.co.uk/ |
| Description | Collaboration with ARC (South Africa) on flow cytometry phenotyping of cattle B cells |
| Organisation | Agricultural Research Council (ARC) |
| Country | South Africa |
| Sector | Public |
| PI Contribution | We hosted a scientist form ARC (TN) to train them in flow cytometry of cattle B cell phenotyping. |
| Collaborator Contribution | None |
| Impact | Transfer of reagents, knowledge and expertise to a LMIC. |
| Start Year | 2023 |
| Description | Development of methods for species switching of mouse monoclonal antibodies to mouse-porcine hybrid recombinant antibody |
| Organisation | Technical University of Denmark |
| Country | Denmark |
| Sector | Academic/University |
| PI Contribution | • Provided mouse and porcine immunoglobulin expression vectors to enable expression and species switching of mouse monoclonal antibodies to mouse-porcine hybrid recombinant antibodies. • Provided intellectual input in designing the recombinant antibody expression system. |
| Collaborator Contribution | Provided own antibodies for sequencing and cloning. |
| Impact | The protocols and reagents that we provided have enabled Prof. Gregers Jungersen group at DTU to amplify and Sequence several important monoclonal antibodies against cell surface molecules of the pig immune cells. These sequences will now enable them to molecularly engineer these antibodies to make mouse-pig hybrid recombinant antibodies as a useful research tool for in vitro investigations of cells and antibody interactions. |
| Start Year | 2018 |
| Description | Development of methods for species switching of mouse monoclonal antibodies to mouse-porcine hybrid recombinant antibody - WM |
| Organisation | Technical University of Denmark |
| Department | Department of Biotechnology and Biomedicine |
| Country | Denmark |
| Sector | Academic/University |
| PI Contribution | • Provided mouse and porcine immunoglobulin expression vectors to enable expression and species switching of mouse monoclonal antibodies to mouse-porcine hybrid recombinant antibodies. • Provided intellectual input in designing the recombinant antibody expression system. |
| Collaborator Contribution | TBA |
| Impact | TBA |
| Start Year | 2018 |
| Description | Development of recombinant antibody production pipeline |
| Organisation | Rosalind Franklin Institute |
| Country | United Kingdom |
| Sector | Charity/Non Profit |
| PI Contribution | Provided Bovine, porcine and chicken immunoglobulin sequences to enable design of recombinant antibody Expression vectors. |
| Collaborator Contribution | • Provided intellectual input in designing recombinant antibody expression system. • Provided backbone vectors for recombinant antibody expression • Provided training for three staff members on recombinant antibody production methods |
| Impact | Using the tools developed, 100 antibodies against cell surface molecules of cattle, pig, chicken immune cells and viruses have already been sequenced and cloned in expression vectors. The first sequencing step in the recombinant workflow has been applied to many more hybridoma cells, so these reagents are now secure for the future as they exist as sequences and are also available to be molecularly engineered to better suit research needs e.g. available to be class switched if required. |
| Start Year | 2018 |
| Description | Development of recombinant antibody production pipeline - WM |
| Organisation | Research Complex at Harwell |
| Department | Oxford Protein Production Facility-UK (OPPF-UK) |
| Country | United Kingdom |
| Sector | Academic/University |
| PI Contribution | Provided Bovine, porcine and chicken immunoglobulin sequences to enable design of recombinant antibody vectors. |
| Collaborator Contribution | • Provided intellectual input in designing recombinant antibody expression system. • Provided backbone vectors for recombinant antibody expression • Provided training for three staff members on recombinant antibody production methods |
| Impact | Using the tools developed, 50 important hybridomas (held at the Pirbright Institute) for monoclonal antibodies against cell surface molecules of cattle, pig and chicken immune cells have already been sequenced and cloned in expression vectors. These reagents are now secure for the future as they exist as sequences and are also available to be molecularly engineered to better suit research needs e.g available to be class switched if required. |
| Start Year | 2018 |
| Description | EBI |
| Organisation | EMBL European Bioinformatics Institute (EMBL - EBI) |
| Country | United Kingdom |
| Sector | Academic/University |
| PI Contribution | We maintain and curate the IPD-MHC database |
| Collaborator Contribution | They host the database and provide generic tools as an in kind contribution |
| Impact | All usage of the website and other outcomes associated with this award |
| Start Year | 2015 |
| Description | Monoclonal antibody sequencing |
| Organisation | University of Edinburgh |
| Department | The Roslin Institute |
| Country | United Kingdom |
| Sector | Academic/University |
| PI Contribution | Performed the sequencing of hybridoma stocks from the Roslin Institute. |
| Collaborator Contribution | Provided intellectual input and hybridoma cells for sequencing. |
| Impact | Using the tools that we have developed, 29 hybridomas of important monoclonal antibodies (from the Roslin Institute) against cell surface molecules of Cattle, pig and chicken immune cells have already sequenced. The sequences will enable securing these reagents for the future as they will exist as sequences and are also make them available to be molecularly engineered to better suit research needs. |
| Start Year | 2018 |
| Description | Monoclonal antibody sequencing - WM |
| Organisation | University of Edinburgh |
| Department | The Roslin Institute |
| Country | United Kingdom |
| Sector | Academic/University |
| PI Contribution | Performed the sequencing of hybridoma stocks from the Roslin Institute |
| Collaborator Contribution | Provided intellectual input and hybridoma cells for sequencing |
| Impact | Using the tools that we have developed, 20 additional hybridomas of important monoclonal antibodies (from the Roslin Institute) against cell surface molecules of Cattle, pig and chicken immune cells have already amplified by PCR and submitted for sequencing. The sequences, when analysed, will enable securing these reagents for the future as they will exist as sequences and are also make them available to be molecularly engineered to better suit research needs. |
| Start Year | 2018 |
| Description | BSI antibody meeting |
| Form Of Engagement Activity | A talk or presentation |
| Part Of Official Scheme? | No |
| Geographic Reach | International |
| Primary Audience | Professional Practitioners |
| Results and Impact | Presented work on how we have developed methods to study cattle B cells and antibody responses. |
| Year(s) Of Engagement Activity | 2018 |
| Description | Gordon's schools careers Fair-AH |
| 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 | Provoking interest in a range of science career paths |
| Year(s) Of Engagement Activity | 2020 |
| Description | Mock Interview Programme Cove School- Ai, AA |
| 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 | Stimulating increased interest in science and research |
| Year(s) Of Engagement Activity | 2019 |
| Description | Sir John Kingman (UKRI) visit |
| Form Of Engagement Activity | Participation in an open day or visit at my research institution |
| Part Of Official Scheme? | No |
| Geographic Reach | National |
| Primary Audience | Policymakers/politicians |
| Results and Impact | Visit to highlight Pirbright science to UKRI |
| Year(s) Of Engagement Activity | 2019 |
| Description | Surrey MSc Student Visit - WM |
| Form Of Engagement Activity | A talk or presentation |
| Part Of Official Scheme? | No |
| Geographic Reach | Local |
| Primary Audience | Postgraduate students |
| Results and Impact | Changes in public attitudes and stimulating increased interest in research |
| Year(s) Of Engagement Activity | 2019 |
| Description | Surrey Vet School Open Day - WM |
| Form Of Engagement Activity | Participation in an activity, workshop or similar |
| Part Of Official Scheme? | No |
| Geographic Reach | National |
| Primary Audience | Undergraduate students |
| Results and Impact | Changes in public attitudes and stimulating increased interest in research |
| Year(s) Of Engagement Activity | 2018 |
| Description | Utrect-EVIC |
| Form Of Engagement Activity | A formal working group, expert panel or dialogue |
| Part Of Official Scheme? | No |
| Geographic Reach | International |
| Primary Audience | Postgraduate students |
| Results and Impact | Sat as part of an expert panel on the future of immune reagents for veterinary research |
| Year(s) Of Engagement Activity | 2018 |