The majority of the UK population and a lesser proportion of continental European and the rest of the world have been exposed to mad cow disease or BSE contaminated foods and this has resulted in the new human disease variant CJD (vCJD). Although the number of patients remains small, the number of people infected but without any signs of illness is unknown and could be very large. The time from being infected to showing signs of the disease can be prolonged for up to 50 years. During this period infected individuals are themselves infectious and there is a risk of spreading the disease through the contamination of medical and dental instruments; the use of contaminated blood for transfusion and the transplantation of infected organs such as kidneys. All of these dangers pose significant challenges for the protection of public health and measures currently in place cost 100’s of millions of pounds per annum in the UK alone.
After infection with BSE there is a characteristic change that occurs to one of the body’s normal proteins in people who have the disease and the detection of this altered protein or PrPSc prion can be used to show someone has CJD. Unfortunately the amounts of prion are very low in blood and are difficult to tell apart from the large amounts of the normal protein that are always present in both healthy and infected people.
We are trying different ways of detecting this rare altered protein which accumulates as ordered aggregates. One approach is to copy the way the prion reproduces itself in the human body during disease. This allows us to ‘amplify’ the very small amounts of prion in blood samples into large amounts that can be easily detected using conventional laboratory tests. In doing so we are also gaining insights into the fundamental processes of prion replication that distinguish this particular protein misfolding event from those involved in other diseases that produce very similar aggregated proteins that do not infect others.
Our research requires interaction with other Unit programmes working to identify genetic modifiers of susceptibility to infection (Dr Simon Mead) and incubation period (Dr Sarah Lloyd). We work closely with Dr Jonathan Wadsworth who is leading research to purify the infectious agent. In the course of his research he is identifying other proteins and macromolecules that are associated with prions and assist us defining the molecular mechanisms of prion replication. Both ourselves and Dr Wadsworth’s group also have close interactions with Professor Anthony Clarke’s team in attempting to apply all of our research to generate synthetic prions and determine their molecular organisation and atomic structure.
Another key interface is between ourselves and Dr Julie Edgeworth who is characterising the extraordinary avidity that prions have for certain materials, in particular stainless steel. This has led to the development of a novel cell-based assay for surface bound prions that is more sensitive than conventional bioassay and is ideal for evaluating reagents and methods for the decontamination of surgical and dental instruments. An additional benefit of this work has been the discovery of effective methods for capturing and concentrating abnormal PrP from blood to enhance detection by immunoassay.
Detection of prion infection in variant Creutzfeldt-Jakob disease: a blood based assay
Edgeworth, J.A., Farmer, M., Sicilia, A., Tavares, P., Beck, J., Campbell, T., Lowe, J., Mead, S., Rudge, P., Collinge, J. and Jackson G.S.
Lancet 2011 377:487-93
A standardised comparison of commercially available prion decontamination reagents using the standard steel binding assay
Edgeworth JA, Sicilia A, Linehan J, Brandner S, Jackson GS, Collinge J.
J Gen Virol. 2010 Nov 17. [Epub ahead of print]
Pharmacological chaperone for the structured domain of human prion protein
Nicoll AJ, Trevitt CR, Tattum MH, Risse E, Quarterman E, Ibarra AA, Wright C, Jackson GS, Sessions RB, Farrow M, Waltho JP, Clarke AR, Collinge J.
Proc Natl Acad Sci USA 2010 Oct 12; 107(41):17610-5
The H187R mutation of the human prion protein induces conversion of recombinant prion protein to PrPSc-like form
Hosszu LL, Tattum MH, Jones S, Trevitt CR, Wells MA, Waltho JP, Collinge J, Jackson GS, Clarke AR.
Biochemistry, 2010, 49, 8729–8738
Spontaneous generation of mammalian prions
Julie A. Edgeworth, Nathalie Gros, Jack Alden, Susan Joiner, Jonathan D. F. Wadsworth, Jackie Linehan,
Sebastian Brandner, Graham S. Jackson, Charles Weissmann, and John Collinge.
Proc Natl Acad Sci USA 2010 Aug 10; 107(32):14402-6
PNRP Alllelic Series from 19 years of prion protein gene sequencing at the MRC Prion Unit
Beck JA, Poulter M, Campbell TA, Adamson G, Uphill JB, Guerreiro R, Jackson GS, Manji H, Collinge J, Mead S.
Hum Mutat. 2010 Jul; 31(7): E1551-63
A highly sensitive immunoassay for the detection of prion-infected material in whole human blood without the use of proteinase K
Tattum MH, Jones S, Pal S, Khalili-Shirazi A, Collinge J, Jackson GS
Transfusion. 2010 Jun 18
Superoxide dismutase 1 and tgSOD1 mouse spinal cord seed fibrils, suggesting a propagative cell death mechanism in amyotrophic lateral sclerosis
Chia R, Tattum MH, Jones S, Collinge J, Fisher EM, Jackson GS.
PLoS One. 2010 May 13;5(5):e10627
The Legs at odd angles (Loa) mutation in cytoplasmic dynein ameliorates mitochondrial function in SOD1G93A mouse model for motor neuron disease
Morsi El-Kadi A, Bros-Facer V, Deng W, Philpott A, Stoddart E, Banks G, Jackson GS, Fisher EM, Duchen MR, Greensmith L, Moore AL, Hafezparast M
J Biol Chem. 2010 Jun 11;285(24):18627-39
Modification of superoxide dismutase 1 (SOD1) properties by a GFP tag - implications for research into amyotrophic lateral sclerosis (ALS).
Stevens JC,Chia R, Hendriks WT, Bros-Facer V,van Minnen J, Martin JE,Jackson GS,Greensmith L,Schiavo G, Fisher EM
PLoS One. 2010 Mar 8;5(3):e9541
Discrimination between prion-infected and normal blood samples by protein misfolding cyclic amplification
Tattum MH, Jones S, Pal S, Collinge J, Jackson GS
Transfusion. 2010 May;50(5):996-1002
The R1441C mutation alters the folding properties of the ROC domain of LRRK2
Li Y, Dunn L, Greggio E, Krumm B, Jackson GS, Cookson MR, Lewis PA, Deng J.
Biochim Biophys Acta. 2009 Dec;1792(12):1194-7
Conformational properties of beta-PrP
Hosszu LL, Trevitt CR, Jones S, Batchelor M, Scott DJ, Jackson GS, Collinge J, Waltho JP, Clarke AR.
J Biol Chem. 2009 Aug 14;284(33):21981-90
Crystal structure of human prion protein bound to a therapeutic antibody.
Antonyuk, S.V., Trevitt, C., Strange, R., Jackson, G.S., Sangar, D., Batchelor, M., Cooper, S., Fraser, C., Khalili-Shirazi, A., Clarke, A.R., Hasnain, S.S. and Collinge, J.
Proc Natl Acad Sci U S A. 2009 Feb 24;106(8):2554-8
Folding kinetics of the human prion protein probed by temperature jump.
Hart, T.C., Hosszu, L.L.P., Trevitt, C.R., Jackson, G.S., Waltho, J.P., Collinge, J., Clarke, A.R.
Proc Natl Acad Sci U S A. 2009 Apr 7;106(14):5651-6
Highly sensitive, quatative cell-based assay for prions adsorped to solid surfaces.
Edgeworth, J.A., Jackson, G.S., Clarke, A.R., Weissman, C. and Collinge J.
Proc Natl Acad Sci U S A. 2009 Mar 3;106(9):3479-83
A role of cellular prion protein in programming T-cell cytokine responses in disease.
Ingram, R.J., Isaacs, J.D., Kaur, G., Lowther, D.E., Reynolds, C.J., Boyton, R.J., Collinge, J., Jackson, G.S. and Altmann, D.M.
The FASEB Journal 2009 Jun;23(6):1672-84
The cellular prion protein is preferentially expressed by CD4+ CD25+ Foxp3+ regulatory T-cells.
Isaacs, J.D., Garden, O.A., Kaur, G., Collinge, J., Jackson, G.S., and Altmann, D.M.
Journal of Immunology 125(3):313-9 (2008).
Detection of proteinase K-sensitive disease related prion protein with thermolysin.
Cronier, C., Gros, N., Tattum, M.H., Jackson, G.S., Collinge, J. and Wadsworth, J.D.F.
Biochemical Journal 416(2):297-305 (2008).
Inhibition of proteinase K activity by copper(II) ions.
Stone, L.A., Jackson, G.S., Clarke, A.R., Collinge, J. and Wadsworth, J.D.
Biochemistry 46(1):245-252 (2007).
Efficient dissemination of prions through preferential transmission to nearby cells.
Pacquet, S., Langevin, C., Chapius, J., Jackson, G.S., Laude, H. and Vilette, D.
Journal of General Virology 88: 706-713 (2007).
Disease-associated prion protein oligomers inhibit the 26S proteasome.
Kristiansen, M., Deriziotis, P., Dimcheff, D.E., Jackson, G.S.,Ovaa, H.,.Naumann, H., Clarke, A.R., van Leeuwen, F.W, Menéndez-Benito, V., Dantuma, N.P., Portis, J.P., Collinge, J. and Tabrizi, S.J.
Mol. Cell. 26(2):175-88 (2007).
Proteomic Profiling of Plasma in Huntington's Disease Reveals Neuroinflammatory Activation and Biomarker Candidates.
Dalrymple A., Wild E.J., Joubert R., Sathasivam K., Bjorkqvist M., Petersen A., Jackson G.S., Isaacs J.D., Kristiansen M., Bates G.P., Leavitt B.R., Keir G., Ward M., Tabrizi S.J.
J. Proteome Res. 6(7):2833-40 (2007).
The b-PrP form of the human prion protein stimulates production of monoclonal antibodies to epitope 91-110 that recognise native PrPSc.
Khalili-Shirazi, A, Kaisar, M., Mallinson, G., Jones, S., Bhelt, D., Fraser, C., Clarke, A.R., Hawke, S.H., Jackson, G.S. and Collinge, J.
Biochim Biophys Acta 1774(11):1438-50 (2007).
Elongated Oligomers assemble into mammalian amyloid fibrils. Tattum, M.H., Cohen-Krausz, S., Khalili-Shirazi, A., Jackson, G.S., Orlova, E, Collinge, J., Clarke, A.R. and Saibil, H.
Journal of Molecular Biology 357(3):975-85 (2006).
The prion protein does not require a GPI anchor to be infectious.
Lewis, P.A., Properzi, F., Prodromidou, P., Clarke, A.R., Collinge, J. and Jackson, G.S.
Biochemical Journal 395(2):443-8 (2006).
Codon 129 polymorphism of the human prion protein influences the kinetics of amyloid formation.
Lewis, P., Tattum, M.H., Jones, S., Bhelt, D., Batchelor, M., Clarke, A.R., Collinge, J. and Jackson, G.S.
Journal of General Virology 87: 2443-2449 (2006).
A Quantitative Reassessment of Copper (II) Binding in the Full-Length Prion Protein.
Wells, M.A., Jackson, G.S., Jones, S., Hosszu, L.L.P., Craven, C., Clarke, A.R., Collinge, J. and Waltho, J.P.
Biochemical Journal 399(3):435-44 (2006).
Multiple forms of copper(II) coordination occur throughout the disordered N-terminal region of the prion protein at pH7.4.
Wells, M.A., Jelinska, J., Hoszu, L.L.P., Craven, J.C., Clarke, A.R., Collinge, J., Waltho, J.P. and Jackson, G.S.
Biochemical Journal 400(3) 501-510 (2006).
The human prion protein residue 129 polymorphism lies within a cluster of epitopes for T cell recognition.
Isaacs, J.D., Collinge, J., Altmann, D.M. and Jackson G.S.
Journal of Neuropathology and Experimental Neurology 65(11):1059-1068 (2006).
The role of the cellular prion protein in the immune system.
Isaacs JD, Jackson GS, Altmann DM.
Clin Exp Immunol. 2006 Oct;146(1):1-8
An enzyme-detergent method for effective prion decontamination of surgical steel.
Jackson, G.S., Mackintosh, E., Flecgsig, E., Prodromidou, K., Hirsch, P., Linehan, J., Brandner, S., Clarke, A.R., Weissman, C. and Collinge, J.
Journal of General Virology 86(3): 869-878 (2005).
Protein conformation significantly influences immune responses to prion protein.
Khalilli-Shirazi, A., Summer, L., Londei, M., Quarantino, S., Clarke, A.R., Hawke, S., Jackson, G.S. and Collinge J.
Journal of Immonology 174(6): 3256-3263 (2005).
PrP Glycoforms are associated together in a strain-specific ratio in native PrPSc.
Khalilli-Shirazi,A., Summers, L., Linehan, J., Mallinson, G., Anstee, D., Hawke, S., Jackson, G.S., and Collinge J.
Journal of General Virology 86(9): 2635-2644 (2005).
Recombinant prion protein does not possess SOD-1 activity.
Jones, S., Batchelor, M., Bhelt, D., Clarke, A.R., Collinge J. and Jackson, G.S.
Biochemical Journal 392(2), 309-312 (2005).
Defineable equilibrium states in the folding of human Prion protein.
Hosszu, L.L.P., Wells, M.A., Jackson, G.S., Jones, S., Batchelor, M., Clarke, A.R., Craven, C.J., Waltho, J.P. and Collinge, J.
Biochemistry 44 (50), 16649-16657 (2005).