Molecular analysis of the lymphocyte cell surface

Lead Research Organisation: University of Oxford
Department Name: Sir William Dunn Sch of Pathology

Abstract

My first aim is to continue to ensure that our work is published as soon as possible in mainstream journals and in the form of reviews to meet a wider audience. The WWW is becoming very important in disseminating information and our WWW pages are regularly updated. I am keen on open access and I have contributed to the recent government enquiry on this subject. I have recently published an article in BMC Biochem with another to submitted later this month. The nature of our basic research is relatively technical and not disease oriented which limits interest to the media. However in the past press releases have led to TV and radio interest and I will continue to encourage these where appropriate. I will continue to take the opportunity to discuss the work and recently I was the focus for an article in the magazine Genome Technology (Feb 2003). We will continue to encourage visits from school members and one visit to a school is planned shortly (MHB). In addition two members of my group presented at the recent British Association for the Advancement of Science meeting in Manchester along with a former Post doc and this type of contact will be encouraged.

Technical Summary

The immune system provides a system of cells that provide a highly controlled response of the optimal duration at the site of the pathogen and very rarely makes errors but in these circumstances they lead to autoimmune disease or cancer. It is through the interactions of these membrane molecules that the immune system is regulated and the characterisation of these interactions at a molecular level is central to understanding the immune system and improving therapies for autoimmune diseases, cancer and permitting transplantation. The programme continues the identification of these interactions and use of the resulting reagents to analyse their functions at a molecular level. The interactions are characterised quantitviely as this is essential in working out the relevant interactions in mediating fucntion. The major lines of research are;
1. The systematic identification of interactions of T cell membrane proteins using existing methods and new protein microarray assays.
2. The continuing detailed biochemical analysis of CD200/CD200R, CD47/SIRP and CD6/CD166 interactions that have been shown to regulate immune cells through cell cell contact
3. Viral homologues of CD200 and CD47 will be characterised to understand their role in the virus and of the host protein.
4. Proteins interacting with the cytoplasmic regions of the above receptors will be identified biochemically giving insight into the mode of transmission of signals.
5. Model systems of immune reactions will be perturbed in vivo and in vitro using antibodies and recombinant proteins and changes in gene expression characterised to probe functional effects.
6. Production of the extracellular regions of these proteins using cell lines with reduced glycosylation heterogeneity for X-ray crystallography using high throughput crystallisation methods at the MRC Oxford Protein Production Facility
7. Further detailed functional analysis will be carried out in collaboration with academic and industrial partners and potential as therapeutics explored.

Publications

10 25 50
 
Description MRC Research Grant
Amount £1,800,000 (GBP)
Organisation Medical Research Council (MRC) 
Sector Academic/University
Country United Kingdom
Start 04/2010 
End 03/2015
 
Description MRC Research Grant
Amount £190,000 (GBP)
Organisation Medical Research Council (MRC) 
Sector Academic/University
Country United Kingdom
Start 07/2007 
End 07/2010
 
Description Wellcome Trust Project Grant (The structural basis of the binding specificity of the family of SIRP paired receptors)
Amount £210,000 (GBP)
Funding ID 084194 
Organisation Wellcome Trust 
Sector Charity/Non Profit
Country United Kingdom
Start 04/2008 
End 04/2011
 
Title Monoclonal antibodies 
Description Monoclonal antibodies have been made that recongise membrane proteins. They are vital for our research but we continue to make them widely available to the scientific community by direct distribution and through commercialisation. 
Type Of Material Antibody 
Year Produced 2007 
Provided To Others? Yes  
Impact They allow proteins to be recognised. Equivalent reagents have not been produced by others 
 
Title Monoclonal antibody 
Description Monoclonal antibody 
Type Of Material Antibody 
Provided To Others? No  
Impact gives new insight into family of receptors 
 
Title Multivalent reagents to detect weak protein interactions 
Description We developed new ways to detect the weak protein interactions that are common for cell surface proteins but difficult to detect. 
Type Of Material Data analysis technique 
Provided To Others? No  
Impact We identified one particuarly interesting interaction and we and others are using the technique. 
 
Description Analysis of viral homologues 
Organisation Robert Koch-Institut
Country Germany 
Sector Public 
PI Contribution We analysed the reaction between a viral protein and the host receptor. Viral work carried out by collaborator.
Collaborator Contribution On going collaboration that should lead to publishable results
Impact Publication 19570854. Continuing to follow up results under curent programme grant
 
Description Control of dendritic cell activity 
Organisation Technical University of Dresden
Country Germany 
Sector Academic/University 
PI Contribution We provided reagents and scientific input
Collaborator Contribution They did the majority of the functional experiments
Impact 16782032
 
Description Interactions with extracellular matrix 
Organisation Cleveland Clinic
Department Department of Cellular & Molecular Medicine
Country United States 
Sector Academic/University 
PI Contribution We carried out BIAcore studies that complemented their studies
Collaborator Contribution The group are experts in extracelullar matrix and able to allow us to relate our biochemical data to their structure and functional data
Impact 18285447
Start Year 2007
 
Description Role of CD200/CD200R in viral infection 
Organisation University of Oxford
Department Kennedy Institute of Rheumatology
Country United Kingdom 
Sector Academic/University 
PI Contribution We provided reagents and data about biochemical analysis vital for interpretation
Collaborator Contribution It allowed expansion of our studies on CD200/CD200R to be expanded into the area of viral infection
Impact 18660812
 
Description Structure of CD5 
Organisation Medical Research Council (MRC)
Department MRC National Institute for Medical Research (NIMR)
Country United Kingdom 
Sector Public 
PI Contribution We made recombinant proteins, tested mutants made by NIMR and advised on significance
Collaborator Contribution This collaborator solved the structure of protein involved in T cell recognition. This was technically very difficult and the successful approach was reached after many attempts and more than 10 years.Our collaborator solved the NMR structure of CD5 that we study. This was technically very difficult and many appraches were attempted over more than 10 years
Impact 18339402
 
Description Structure of CD5 
Organisation Medical Research Council (MRC)
Department MRC National Institute for Medical Research (NIMR)
Country United Kingdom 
Sector Public 
PI Contribution We made recombinant proteins, tested mutants made by NIMR and advised on significance
Collaborator Contribution This collaborator solved the structure of protein involved in T cell recognition. This was technically very difficult and the successful approach was reached after many attempts and more than 10 years.Our collaborator solved the NMR structure of CD5 that we study. This was technically very difficult and many appraches were attempted over more than 10 years
Impact 18339402
 
Description X-ray crystallography of membrane proteins 
Organisation University of Oxford
Department Sir William Dunn School of Pathology
Country United Kingdom 
Sector Academic/University 
PI Contribution A new collaboration has now been set up with Susan Lea's group in Pathology. One structure has been solved and another is on the way (no published)
Collaborator Contribution This new collaboration has now been set up with Susan Lea's group in Pathology. One structure has been solved and another is on the way (not published)
Impact One structure has been solved and another is on the way (no published). The collaboration allows direct access to X-ray crystallography with one member of my group working at alls tages
Start Year 2009
 
Description X-ray crystallography of membrane proteins 
Organisation University of Oxford
Department Wellcome Trust Centre for Human Genetics
Country United Kingdom 
Sector Academic/University 
PI Contribution We made all the proteins, set up crystallisation trials using faciliteis at Wellcome Trust Centre. They solved the crystal structures and helped in their analysis.
Collaborator Contribution We had a joint project that led to high profiile publications
Impact 19628875 18657508 17369261
 
Description Talk to West Oxford Academy by group member 
Form Of Engagement Activity A talk or presentation
Part Of Official Scheme? Yes
Primary Audience Public/other audiences
Results and Impact Talk and discussion to local group in Oxford

Considerable interest and postive feedback of interest
Year(s) Of Engagement Activity 2008