Identification of HLA-B*27 bound peptides involved in the pathogenesis of Ankylosing Spondylitis and Reactive Arthritis

Lead Research Organisation: University of Oxford
Department Name: Target Discovery Institute


HLA-B*27 is found in over 90% of patients with Ankylosing Spondylitis and Reactive Arthritis, two of the most common inflammatory arthritic diseases. Yet this immune protein is present in only 5-7% of people without these diseases. HLA-B*27, like other HLA types, is present on almost all cells of the body. The normal function of HLA-B*27 is to bind to foreign peptides that come from infecting microbes, and to stimulate T cells to eliminate the infection. One hypothesis is that sometimes T cells that are stimulated by foreign microbes also react to peptides bound to HLA-B*27 that come from normal, uninfected cells. This is a theory we want to test. In this study, we will take T cells from the joints of patients with inflammatory arthritis and use a powerful, new method to identify all the peptides they respond to. The technique involves putting HLA-B*27 onto yeast cells, each of which can place a different peptide onto the HLA-B*27 protein. We can make 100 million yeast cells, each with a different peptide. Then we will make the receptor proteins, namely the T cell receptors (TCRs), that normally react with HLA-B*27 plus peptide, from the patient's T cells, to select the yeast cells that bind. Finally, we will analyse the HLA-B*27 bound peptides that the TCRs recognise and compare them to all the peptides in the human and microbe databases. The latter have been accumulated from the Human Genome Project and the Microbe Genome Project over the last 10 years. This will allow us to identify both the microbe peptides that first stimulated the T cells, and the peptides from normal cells that they potentially attack to cause arthritis. The ultimate aim of this study is to identify if peptides that bind to HLA-B*27 cause disease. If we are successful we, and others, should be able to make candidate treatments that interfere with that process. This approach to therapy will be highly specific to this type of arthritis and far less likely to cause serious side effects than current treatments.

Technical Summary

Although the link between Ankylosing Spondylitis (AS)/Reactive Arthritis (ReA) and HLA-B*27 has been recognised for 40 years, the pathogenic role played by the disease-associated MHC class I subtypes is not understood. Various theories have been proposed, including models implicating the unusual biology of HLA-B*27 that drive the production of aberrant protein forms which contribute to inflammatory disease. The alternative and original, 'arthritogenic peptide' model suggests that disease pathology relates to normal HLA-B*27 forms, where pathogen-specific HLA-B*27-restricted T cells recruited during natural infection subsequently cross-react with self-peptides bound to HLA-B*27 and promote autoimmunity. Recent GWAS studies have re-ignited interest in this latter model, but the lack of powerful techniques to perform peptide-hunting screens have severely hampered progress in this area. In this study, we want to apply newly developed, cutting-edge MHC class I-peptide yeast display technology to re-appraise the 'arthritogenic peptide' model of disease. For the first time, this approach offers a non-bias platform to hunt for peptides that might be linked with disease. Used in conjunction with disease-associated T cell receptor screening tools, the HLA-B*27 yeast platforms will allow high-throughput screening experiments to be performed. Our goal is that this study should dissect the relevance of this model in relation to disease pathology, whilst also driving discoveries amenable to therapeutic development.

Planned Impact

A number of different sectors stand to benefit from this work.

It is anticipated that this study could lead to the development of new therapies that improve treatments for SpA. Current treatments for inflammatory arthritis, such as non-steroidal anti-inflammatory agents (NSAIDs) and anti-TNF therapies, can cause side effects; they work broadly and down-stream of the initiating disease trigger. If our research demonstrates that specific T cells recognise peptides that cause autoimmunity, then newer therapies that precisely target these cells could be developed. Improved therapies could also lead to an economic benefit. This would be experienced by pharmaceutical companies who steer the development of these reagents. An economic benefit could also occur in the workplace, with new and improved treatments leading to a reduction in patient morbidity, and therefore absenteeism.

If our work sheds light on the fundamental biology of inflammatory arthritis, not only will this benefit our scientific peers, but it will also improve the accuracy and breath of biological information available to second and third level science/medical students. Findings in this area should add to our overall understanding of general immunology.

Findings from our research could also inform funding bodies, including grant councils, how to best invest future funds to study inflammatory arthritis. Up to three models linking HLA-B*27 to inflammatory disease currently exist - if our research demonstrates that the disease model under investigation in this proposal is primarily involved in disease pathogenesis, then this information could inform subsequent government or charity funding initiatives (for example, via the initiation of funding 'calls' specific to this). Our study could also help guide treatments approved by the NHS, to devise regimens that are most beneficial and cost-effective.


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Description AJD00080 AJ01.01 T cell-based diagnostic assay development for Ankylosing Spondylitis
Amount £7,000 (GBP)
Funding ID AJD00080 AJ01.01 
Organisation Oxford University Press 
Sector Private
Country United Kingdom
Start 05/2019 
End 07/2021
Description MRC Project Grant
Amount £500,000 (GBP)
Organisation Medical Research Council (MRC) 
Sector Public
Country United Kingdom
Start 06/2015 
End 06/2018
Description Rosetrees Trust
Amount £21,000 (GBP)
Organisation Rosetrees Trust 
Sector Charity/Non Profit
Country United Kingdom
Start 03/2015 
End 03/2018
Description Rosetrees Trust matched-funding stipend
Amount £15,000 (GBP)
Organisation Rosetrees Trust 
Sector Charity/Non Profit
Country United Kingdom
Start 03/2018 
End 10/2018
Title Blue Native analysis of MHC 
Description We have applied Blue Native gel technology to the analysis of proteins - this approach allows to evaluate the conformational heterogeneity and stability of proteins samples. We have deciphered that the technique is especially useful for mac class I proteins. 
Type Of Material Technology assay or reagent 
Year Produced 2018 
Provided To Others? Yes  
Impact Colleagues have asked or commented on the utility of the approach 
Description Dmitriy TCRs 
Organisation Masaryk University
Country Czech Republic 
Sector Academic/University 
PI Contribution We have generated a soluble T cell receptor screening reagent based in the information supplied by this collaborator.
Collaborator Contribution The collaborators provided the sequences of the T cell receptor chain from which we generated the soluble disease-related T cell receptor
Impact Unfortunately, this TCR did not retrieve any hits in the HLA-B*27 screening runs. However, the collaboration remains open for further evaluation of disease-related T cell receptors that might emerge in subsequent screens. Disciplines includes T cell receptor identification (sequencing), production of soluble T cell receptor reagents (protein production) and screening by HLA*B27 yeast display (molecular and sequencing).
Start Year 2017
Description Inman TCRs 
Organisation University of Toronto
Country Canada 
Sector Academic/University 
PI Contribution We are generating disease related T cell receptor reagents based on their lab-based findings
Collaborator Contribution Their are providing us with T cell receptor sequence which allows us to generate disease related T cell receptor reagents
Impact Outcomes will depend whether these candidate T cell receptors lead to the identification of pathogen and self related antigens that underlie the pathogenesis of HLA-B*27-related SpA. Should they be identified, they is potential for the development of antigen-based therapeutics that specifically block these T cells. Disciplines involved include T cell isolation, generation of soluble TCR reagents (protein production and reagent generation), screening of reagents for potential recognition of self and pathogen-related peptides (molecular screening).
Start Year 2017
Description Professor Chris Garcia 
Organisation Stanford University School of Medicine
Country United States 
Sector Academic/University 
PI Contribution We have provided and produced TCRs from AS patients that are in current screening runs in their MHC yeast display platforms
Collaborator Contribution Our collaborators screens TCRs we have identified in AS patients on their MHC yeast display platforms
Impact Collaboration is ongoing. Initial results published in the journal Nature in late 2022.
Start Year 2015
Description TCR sequences from Synovial fluid - AS study 
Organisation Russian Academy of Sciences
Department Institute of Bio Organic Chemistry, Russia
Country Russian Federation 
Sector Academic/University 
PI Contribution We used the T cell receptor sequence data to prepare T cell receptor screening tools that were run on MHC yeast display screening platforms in Stanford
Collaborator Contribution The group provided disease-linked T cell receptor single cell sequence data from AS patients
Impact Manuscript will be prepared later this year.
Start Year 2018
Description Host to A-level students 
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 Pitched the science to explain how we are using TCRs as tools to hunt for peptide ligands. The students also got hands-on experience that helped towards their UCAS applications, and all were accepted into their 1st choice places at University (Medicine at Imperial, Biology at UCL, and Neuroscience at Bristol)
Year(s) Of Engagement Activity 2014,2016,2017
Description sign-up to In2ScienceUK A-level placement scheme 
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 Aim is to provide students with hands on science experience during placements, in addition to more desk based research (analysis of existing research).

We are planning to hosting students individually, or space allowing as a small group.
Year(s) Of Engagement Activity 2020