Infectious Diseases

Research

Research in the infectious disease unit focuses on persistent human virus infections covering a range of viral pathogens of importance to man, but in particular human herpesviruses and HIV-1. Studies include the analysis of viral assembly, viral-cell interactions and control of latency and reactivation together with the studies on the interactions of viruses with the immune system.

 The central theme running through the research in Professor Lever’s laboratory is the importance of RNA structures and RNA protein interaction in virus assembly. This fundamental paradigm underpins a range of research studies from basic chemistry , biochemistry and structural biology of RNA recognition signals through the cellular pathways which these signals allow viral products to traverse and these are translated ultimately into practical studies of inhibiting viral processes by targeting these regions and subverting them by the development of viral vectors based on lentiviruses.

The diversity of RNA structures engendered by the flexibility and versatility of the RNA molecule has been used by many viral and cellular processes. Unlike proteins, RNA molecules can, in the course of their life adopt many different structures which have different functions and, thus, interact with multiple ligands within the cell, including, of course, their coding potential as messenger RNA. Professor Lever’s interests relate to:- 1. the identification of novel RNA structures within viruses; 2. the cis-acting signals used by viruses in cells to traffic their genomes for viral assembly; 3. the potential therapeutic spin offs of this knowledge in terms of antiviral agents and viral vectors.

Professor John Sinclair’s research is directed at understanding the molecular biology and pathogenesis of human herpes viruses, particularly human cytomegalovirus (HCMV) which is a major cause of disease in transplant patients and patients with AIDS.

As with all herpesviruses, HCMV persists for the lifetime of the host remaining latent in some cells in the infected individual. The mechanism of how latency is maintained and how virus can reactivate is the major goal of his work.

Once reactivated, HCMV can override many cellular control mechanisms to ensure efficient productive infection. Professor Sinclair’s group is also interested how the virus “hijacks” the cell to optimise it for virus production by regulating cell cycle, cellular transcription and apoptosis.

 

Dr Matthew Reeves’ (MRC Career Development Fellow) research programme concerns the mechanism by which cellular changes associated with differentiation and inflammation result in reactivation of latent virus and how viral genes expressed during latency are involved in this switch in viral life cycle.

Professor Sissons and Dr Wills’s laboratory focuses on understanding the balance between immune activation of T cells and NK cells following Human cytomegalovirus (HCMV) infection and the ability of the virus to express a wide array  of immune evasion genes.

HCMV remains an important human pathogen, the molecular immunology of this large human DNA virus informs virus pathogenesis more generally.  An understanding of viral pathogenesis at the level of gene products should lead to rational methods of therapeutic intervention. In addition, studies of the interaction of viral gene products with the host response may lead to novel antiviral strategies. HCMV is unusual in encoding multiple T and NK cell evasion strategies which target multiple mechanisms in antigen processing and presentation, NK activation and inhibition, an understanding of these mechanism should inform us about the basic principles by which cell mediated immune function attempts to overcome intracellular pathogens infecting host cells more generally.

Selected References

  • Bennett NJ, Ashiru O, Morgan FJ, Pang Y, Okecha G, Eagle RA, Trowsdale J, Sissons JG, Wills MR (2010) Intracellular sequestration of the NKG2D ligand ULBP3 by human cytomegalovirus J Immunol Jul 15; 185(2): 1093-102
  • Ashiru O, Bennett NJ, Boyle LH, Thomas M, Trowsdale J, Wills MR NKG2D ligand MICA is retained in the cis-Golgi apparatus by human cytomegalovirus protein UL142 J Virol Dec; 83(23): 12345-54
  • Rizvi TA, Kenyon JC, Ali J, Aktar SJ, Phillip PS, Ghazawi A, Mustafa F, Lever AM (2010) Optimal Packaging of FIV Genomic RNA Depends upon a Conserved Long-range Interaction and a Palindromic Sequence within gag J Mol BiolAug 21. [Epub ahead of print]
  • Reeves M, Woodhall D, Compton T, Sinclair J (2010) Human cytomegalovirus IE72 protein interacts with the transcriptional repressor hDaxx to regulate LUNA gene expression during lytic infection. J Virol Jul; 84(14): 7185-94
  • Li W, Manktelow E, von Kirchbach JC, Gog JR, Desselberger U, Lever AM (2010) Genomic analysis of codon, sequence and structural conservation with selective biochemical-structure mapping reveals highly conserved and dynamic structures in rotavirus RNAs with potential cis-acting functions Nucleic Acids Res Jul 29. [Epub ahead of print]
  • Yang J, Li X, Al-Lamki RS, Southwood M, Zhao J, Lever AM, Grimminger F, Schermuly RT, Morrell NW (2010) Smad-dependent and smad-independent induction of id1 by prostacyclin analogues inhibits proliferation of pulmonary artery smooth muscle cells in vitro and in vivoCirc Res Jul 23; 107(2): 252-62
  • Nejentsev S, Walker N, Riches D, Egholm M, Todd JA (2009) Rare Variants of IFIH1, a Gene Implicated in antiviral responses, protect against type 1 diabetes Science Apr 17; 324(5925): 387-9 link to abstract
  • Poole E, Groves I, MacDonald A, Pang Y, Alcami A, Sinclair J (2009) Identification of TRIM23 as a cofactor involved in the regulation of NF-kappaB by human cytomegalovirus J Virol Apr; 83(8) :3581-90
  • Kenyon JC, Ghazawi A, Cheung WK, Phillip PS, Rizvi TA, Lever AML (2008) The secondary structure of the 5′ end of the FIV genome reveals a long-range interaction between R/U5 and gag sequences, and a large, stable stem-loopRNA Dec;14(12):2597-608
  • Reeves, MB, Davies, AA, McSharry, BP, Wilkinson, GW and Sinclair JH (2007) Complex I binding by a virally encoded RNA regulates mitochondria-induced cell death Science 316: 1345-1348.
  • Goodrum, FD, Reeves, M, Sinclair, J , High, K and Shenk, T (2007) Human cytomegalovirus sequences expressed in latently infected individuals promote a latent infection in vitro Blood in press
  • Waller, EC, McKinney N, Hicks, R, Carmichael AJSissons JG , and Wills MR (2007) Differential co-stimulation through CD137 (4-1BB) restores proliferation of human virus-specific “effector memory” (CD28-CD45RAhi) CD8+ T Cells. Blood
  • Day, EK, Carmichael AJ , ten Berge IJ, Waller EC, Sissons JG , and Wills MR (2007). Rapid CD8+ T Cell Repertoire Focusing and Selection of High-Affinity Clones into Memory Following Primary Infection with a Persistent Human Virus: Human Cytomegalovirus J Immunol 179: 3203-3213
  • Poole, E, King, C, Sinclair, J and Alcami, A (2006) The UL144 gene product of human cytomegalovirus activates NF_B via a TRAF6-dependent mechanism. EMBO J. 18: 4390-4399
  • Anderson, EC, Lever, AML (2006) HIV-1 Gag polyprotein modulates its own translation Journal of Virology ; 80 10478-10486
  • Poole, E, E, Strappe, P, Mok, H-P, Hicks, R, Lever, AML (2005) HIV-1 Gag-RNA interaction occurs at a perinuclear/centrosomal site; analysis by confocal microscopy and FRET Traffic 6: 741-55
  • Zhao, J, Pettigrew, GJ, Bolton, EM, Murfitt, CR, Carmichael, AJBradley, JA, Lever AML (2005) Lentivirus-mediated gene transfer of viral interleukin delays but does not prevent cardiac allograft rejection. Gene Therapy 12: 1509-1516
  • Matthew Reeves, Paul MacAry, Paul LehnerPatrick Sissons and John Sinclair (2005) Latency, Chromatin Remodelling and Reactivation of Human Cytomegalovirus in the Dendritic Cells of Healthy Carriers Proceedings of the National Academy of Sciences (USA) 102: 4140-4145
  • Zeffman, A, Hassard, S, Varani G, Lever AML (2000) The major HIV-1 packaging signal is an extended bulged stem loop whose structure is altered on interaction with the Gag polyprotein J Mol Biol 297: 877-893