Partners: The Pirbright Institute

Staff: Prof. Terry Jackson, Dr. Toby Tuthill, Dr. Don King, Dr. Lidia Lasecka, Dr. Sarah Gold

The Pirbright Institute (formerly the Institute for Animal Health) contains internationally recognised expertise in virology, immunology, genetics, pathogenesis, molecular epidemiology, diagnostics, mathematical biology and bioimaging. Significant investment has been made to redevelop the Pirbright site and the construction and upgrading of facilities is well underway. The inter-disciplinary nature of the institute provides an environment where basic and applied science is combined so fundamental discoveries can be developed with in-house expertise in global disease control and the commercialisation of science.

The Institute specialises in the control of viral diseases and is the only site in the UK licensed for work with foot-and-mouth disease virus (FMDV). Dr Terry Jackson, Dr Toby Tuthill and Dr Don King have combined interests which include molecular mechanisms of FMDV entry, replication and assembly; viral population diversity and evolution; molecular epidemiology and diagnostics.

Toby Tuthill’s research focuses on understanding fundamental molecular mechanisms of picornavirus (FMDV and related viruses) membrane penetration, replication, capsid assembly and genome packaging and the application of this knowledge towards novel targets/strategies for disease control. We use a variety of approaches such as: cell culture, molecular biology, fluorescent microscopy, recombinant proteins, model membranes and electron microscopy. Structural biology approaches such as X-ray crystallography and nuclear magnetic resonance are carried out in collaboration with universities such as Oxford and Harvard.

Current projects include research to answer the following questions: How does FMDV interact with membranes? What is the role of picornavirus capsid protein VP4 in membrane penetration? How does myristoylation and proteolytic processing control the properties of FMDV pentamers? What confers specificity to picornavirus packaging?

Selected publications:

A plate-based high-throughput assay for virus stability and vaccine formulation. Walter TS, Ren J, Tuthill TJ, Rowlands DJ, Stuart DI, Fry EE. J Virol Methods. 2012 Oct;185(1):166-70. Epub 2012 Jun 26.

Crystal structure of equine rhinitis A virus in complex with its sialic acid receptor. Fry EE, Tuthill TJ, Harlos K, Walter TS, Rowlands DJ, Stuart DI. J Gen Virol. 2010 Aug;91(Pt 8):1971-7. Epub 2010 Apr 28.

Picornaviruses. Tuthill TJ, Groppelli E, Hogle JM, Rowlands DJ. Curr Top Microbiol Immunol. 2010;343:43-89.

Cell entry of the aphthovirus equine rhinitis A virus is dependent on endosome acidification. Groppelli E, Tuthill TJ, Rowlands DJ. J Virol. 2010 Jun;84(12):6235-40. Epub 2010 Apr 7.

Equine rhinitis A virus and its low pH empty particle: clues towards an aphthovirus entry mechanism? Tuthill TJ, Harlos K, Walter TS, Knowles NJ, Groppelli E, Rowlands DJ, Stuart DI, Fry EE. PLoS Pathog. 2009 Oct;5(10):e1000620. Epub 2009 Oct 9.

Foot-and-mouth disease virus assembly: processing of recombinant capsid precursor by exogenous protease induces self-assembly of pentamers in vitro in a myristoylation-dependent manner. Goodwin S, Tuthill TJ, Arias A, Killington RA, Rowlands DJ. J Virol. 2009 Nov;83(21):11275-82. Epub 2009 Aug 26.

Recombinant VP4 of human rhinovirus induces permeability in model membranes. Davis MP, Bottley G, Beales LP, Killington RA, Rowlands DJ, Tuthill TJ. J Virol. 2008 Apr;82(8):4169-74. Epub 2008 Feb 6.

Mouse models of rhinovirus-induced disease and exacerbation of allergic airway inflammation. Bartlett NW, Walton RP, Edwards MR, Aniscenko J, Caramori G, Zhu J, Glanville N, Choy KJ, Jourdan P, Burnet J, Tuthill TJ, Pedrick MS, Hurle MJ, Plumpton C, Sharp NA, Bussell JN, Swallow DM, Schwarze J, Guy B, Almond JW, Jeffery PK, Lloyd CM, Papi A, Killington RA, Rowlands DJ, Blair ED, Clarke NJ, Johnston SL. Nat Med. 2008 Feb;14(2):199-204. Epub 2008 Feb 3.

Inhibition of biofilms associated with dentures and toothbrushes by tetrasodium EDTA. Devine DA, Percival RS, Wood DJ, Tuthill TJ, Kite P, Killington RA, Marsh PD. J Appl Microbiol. 2007 Dec;103(6):2516-24.

Inhibition of hepatitis C virus p7 membrane channels in a liposome-based assay system. StGelais C, Tuthill TJ, Clarke DS, Rowlands DJ, Harris M, Griffin S. Antiviral Res. 2007 Oct;76(1):48-58. Epub 2007 Jun 4.

Characterization of early steps in the poliovirus infection process: receptor-decorated liposomes induce conversion of the virus to membrane-anchored entry-intermediate particles. Tuthill TJ, Bubeck D, Rowlands DJ, Hogle JM. J Virol. 2006 Jan;80(1):172-80.

Discrimination among rhinovirus serotypes for a variant ICAM-1 receptor molecule. Xiao C, Tuthill TJ, Bator Kelly CM, Challinor LJ, Chipman PR, Killington RA, Rowlands DJ, Craig A, Rossmann MG. J Virol. 2004 Sep;78(18):10034-44.

Mouse respiratory epithelial cells support efficient replication of human rhinovirus. Tuthill TJ, Papadopoulos NG, Jourdan P, Challinor LJ, Sharp NA, Plumpton C, Shah K, Barnard S, Dash L, Burnet J, Killington RA, Rowlands DJ, Clarke NJ, Blair ED, Johnston SL. J Gen Virol. 2003 Oct;84(Pt 10):2829-36.

Terry Jackson‘s group carries out novel, fundamental research aimed at improving knowledge of the mechanisms of infection by foot-and-mouth disease virus (FMDV). The principle areas of our research are the mechanisms of FMDV endocytosis and formation of the viral replication complex. Through these studies we hope to gain a better understanding of how FMDV replicates, and to inform applied research for control of this important animal pathogen.

We use integrated approaches combining biochemical, molecular- and cell-biology techniques with reverse genetics and advanced microscopy (confocal- and electron-microscopy) to ask fundamental questions about how virus particles attach to cells, how they are internalized by endocytosis, and how they use host-cell membranes for replication. We are using knowledge of FMDV receptors to improve growth of vaccine strains of the virus in cultured cells and to develop FMDV receptors as ‘universal’ virus-capture ligands for use in diagnostic assays. To achieve replication, FMDV dramatically reorganizes internal cellular membranes to provide specialized sites for formation of viral replication complexes. We also study these events as we wish to determine what triggers membrane rearrangements in infected cells, as well as identify the cellular origin of the replication membranes and what properties make them favourable for replication.

Selected publications:

Berryman S, Brooks E, Burman A, Hawes P, Roberts R, Netherton C, Monaghan P, Whelband M, Cottam E, Elazar Z, Jackson T, Wileman T. FMDV induces autophagosomes during cell entry via a class III PI3K-independent pathway. 2012 J Virol. 86:12940-12953.

Seago J, Jackson T, Doel C, Fry E, Stuart D, Harmsen MM, Charleston B, Juleff N. Characterization of epitope-tagged foot-and-mouth disease virus. 2012 J Gen Virol. 93: 2371-2381.

Lohse L, Jackson T, Bøtner A, Belsham GJ. Capsid coding sequences of foot-and-mouth disease viruses are determinants of pathogenicity in pigs. 2012 Vet Res. May 24;43(1):46.

King, Burman, Gold, Shaw, Jackson and Ferris. Integrin sub-unit expression in cell cultures used for the diagnosis of foot-and-mouth disease. 2011 Vet. Immunol. Immunopathol. 15; 140(3-4):259-65.

Bøtner, Kakker, Barbezange, Berryman, Jackson and Belsham. The capsid proteins from field strains of foot-and-mouth disease virus confer a pathogenic phenotype in cattle on an attenuated, cell culture adapted, virus. 2011 J. Gen. Virol. 92:1141-1151

Robinson, Windsor, McLaughlin, Hope, Jackson and Charleston. Foot-and-mouth disease virus exhibits an altered tropism in the presence of specific immunoglobulins, enabling productive infection and killing of dendritic cells. 2011 J. Virol. 85:2212-23.

Dash, Barnett, Denyer, Jackson, Stirling, Hawes, Simpson, Monaghan and Takamatsu. Foot-and-mouth disease virus only replicates transiently in well-differentiated porcine nasal epithelial cells. 2010 J. Virol. 84:9149 – 9160.

Gold, Monaghan, Mertens and Jackson. A clathrin independent macropinocytosis-like entry mechanism used by bluetongue virus-1 during infection of BHK cells. 2010 PLoS ONE: Research Article, published 29 Jun 2010 10.1371/journal.pone.0011360.

Johns, Berryman, Monaghan, Belsham and Jackson. A Dominant-Negative Mutant of rab5 Inhibits Infection of Cells by Foot-and-Mouth Disease Virus: Implications for Virus Entry. 2009 J. Virol. 83:6247-6256.

Armer, Moffat, Wileman, Belsham, Jackson, Duprex, Ryan and Monaghan. Foot-and-mouth disease virus, but not bovine enterovirus, targets the host cell cytoskeleton, via the non-structural protein 3Cpro 2008 J. Virol. 82:10556-10566.

DiCara, Burman, Clark, Berryman, Howard, Hart, Marshall and Jackson. Foot-and-Mouth Disease Virus Forms a Highly Stable, EDTA-Resistant Complex with Its Principal Receptor, Integrin alpha-v/beta-6: Implications for Infectiousness. 2008 J. Virol. 82:1537-1546.

Burman, Clark, Abrescia, Fry, Stuart and Jackson. Specificity of the VP1 GH Loop of Foot-and-Mouth Disease Virus for alpha-v Integrins. 2006 J. Virol. 80:9798-9810.

Monaghan, Gold, Simpson, Zhang, Weinreb, Violette, Alexandersen and Jackson. The alpha-v/beta-6 integrin receptor for foot-and-mouth disease virus is expressed constitutively on the epithelial cells targeted in cattle. 2005 J. Gen. Virol. 86:2769-2780

Berryman, Clark, Monaghan and Jackson. Early Events in Integrin alpha-v/beta-6 Mediated Cell Entry of Foot-and-Mouth Disease Virus. 2005 J. Virol. 79:8519-8534.

Fry, Newman, Curry, Najjam, Jackson, Blakemore, Lea, Miller, Burman, King and Stuart. Structure of Foot-and-mouth disease virus serotype A1061 alone and complexed with oligosaccharide receptor: receptor conservation in the face of antigenic variation. 2005 J. Gen. Virol. 86:1909-1920.

Monaghan, Cook, Jackson, Ryan and Wileman. The ultrastructure of the developing replication site in foot-and-mouth disease virus-infected BHK-38 cells. 2004 J. Gen. Virol. 85:933-946.

Jackson, Clark, Berryman, Burman, Cambier, Mu, Nishimura and King. Integrin alpha-v/beta-8 Functions as a Receptor for Foot-and-Mouth Disease Virus: Role of the beta-Chain Cytodomain in Integrin-Mediated Infection. 2004 J. Virol. 78:4533-4540.

Jackson, Mould, Sheppard and King. The integrin alpha-v/beta1 is a receptor for foot-and-mouth disease virus. 2002 J. Virol. 76:935-941.

Jackson, Sheppard, Denyer, Blakemore and King. The epithelial integrin alpha-v/beta-6 is a receptor for foot-and-mouth disease virus. 2000 J. Virol. 74:4949-4956.

Fry, Lea, Jackson, Newman, Ellard, Blakemore, Abu-Ghazaleh, Samule, King and Stuart. The Structure and Function of a Foot-and-mouth-disease Virus /oligosaccharide receptor complex. 1999 EMBO 18:543-554.

Jackson, Ellard, Abu-Ghazaleh, Brookes, Blakemore, Corteyn, Stuart, Newman and King. Efficient infection of cells in culture by Type-O foot-and-mouth disease virus requires binding to cell surface heparan sulfate. 1996 J. Virol. 70:5282-5287.

Don King heads the Molecular Characterisation and Diagnostics group at the Pirbright Institute. He has a background in veterinary virology and immunology (University of Leeds, University of California, Davis), and is interested in understanding the processes that drive the evolution of positive-stranded viruses such as FMDV.

He manages the core-sequencing facility at the Pirbright Institute that routinely generates complete genome sequences for FMDV, and (in collaboration with Professor Dan Haydon at the University of Glasgow) has pioneered novel analytical pipelines to apply Next-Generation Sequencing methods (Illuminia) to allow the within sample sequence diversity of FMDV to be monitored.

He holds a BBSRC grant to investigate viral evolution (BB/I014314/1: Beyond the consensus: defining the significance of foot-and-mouth disease viral sequence diversity) and is also a co-investigator on two further BBSRC grants (BB/F009186: Identifying epitopes that induce antibody mediated protection against foot-and-mouth disease using reverse genetics) and (BB/H009302/1: Towards the strategic control of endemic foot-and-mouth disease in Africa:). Work within the group (funded by Defra) develops and applies new molecular technologies for the detection and characterisation of important livestock disease agents such as FMD.

These methods underpin the routine diagnostic work of the World Reference Laboratory for FMD (Under the auspices of the FAO, OIE and EU), and were central to the UK national control activities during the FMD outbreaks in 2007. He also coordinates projects funded by EU-FP7, Food-and Agricultural Organisation of the UN to investigate the maintenance of FMD in countries where disease is endemic (Tanzania, Turkey, Malaysia and China).

Selected publications:

Juleff N., Valdazo-González B., Wadsworth J., Wright C. F., Charleston B., Paton D. J., King D. P. and Knowles N. J. Accumulation of nucleotide substitutions occurring during experimental transmission of foot-and-mouth disease virus. Journal of General Virology (IN PRESS).

Valdazo-González B., Polihronova L., Alexandrov T., Normann P., Knowles N. J., Hammond J. M., Georgiev G. K., Özyörük F., Sumption K. J. Belsham G. J. and King D. P. Reconstruction of the transmission history of RNA virus outbreaks using full genome sequences; foot-and-mouth disease virus in Bulgaria in 2011. PLoS ONE (IN PRESS).

Knowles N. J., He J., Shang Y., Wadsworth J., Valdazo-González B., Onosato H., Fukai K., Morioka K., Yoshida K., Cho I.-S., Kim S.-M., Park J.-H., Lee K.-N., Luk G., Borisov V., Scherbakov A., Timina A., Bold D., Nguyen T., Paton D. J., Hammond J. M., Liu X. and King D. P. (2012) Emergence of Southeast Asian Foot-and-mouth disease viruses in East Asia. Emerging Infectious Diseases. 18 (3): 499-501.

Abdul-Hamid N. F., Firat-Saraç M., Radford A. D., Knowles N. J. and King D. P. (2011) Comparative sequence analysis of representative foot-and-mouth disease virus genomes from Southeast Asia. Virus Genes 43: 41-45.

Abdul-Hamid N. F., Hussein N. M., Wadsworth J., Radford A. D., Knowles N. J. and King D. P. (2011) Phylogeography of foot-and-mouth disease virus types O and A in Malaysia and surrounding countries. Infection, Genetics and Evolution 11: 320-328.

Wright C. F., Morelli M. J., Thébaud G., Knowles N. J. Herzyk P., Paton D. J. Haydon D. T. and King D. P. (2011) Beyond the consensus: dissecting within-host population diversity of foot-and-mouth disease virus using next-generation sequencing. Journal of Virology 85 (5): 2266-2275.