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Viral Immunology Research Group

Human cytomegalovirus (HCMV) is a clinically important pathogen with high prevalence worldwide.

Postgraduate opportunity: We've got a funded PhD available. Come and work with us using proteomics to understand how viruses interact with the immune system, and how this can be used to create better vaccines. Find out more and apply now.

Cytomegalovirus

Human cytomegalovirus (HCMV) is a clinically important pathogen with high prevalence worldwide. It is the leading infectious cause of congenital malformation, is associated with life-threatening disease in immunocompromised individuals (e.g. AIDS sufferers and transplant recipients), and is a causative agent of hepatitis, colitis, post-transplantation arteriosclerosis and infectious mononucleosis. As a result, the US Institute of Medicine has designated HCMV as a highest priority (Level I) vaccine target.

We have major interests in the basic biology of the virus, the development of therapeutics and diagnostics, the way in which the virus interacts with the immune system, and using the virus to understand how the immune system functions in both healthy and diseased states.

Discover more about the research carried out into Infection and Immunity.

Research

Immune evasion by HCMV

As a herpes virus, primary infection with HCMV is followed by lifelong persistence during which the immune systems must act to limit the consequences of infection. Through co-evolution, HCMV has developed an intimate relationship with our immune system, and the virus has been established as a paradigm for viral immune evasion.

Natural Killer (NK) cells are implicated in both the innate and adaptive immunity and make a crucial contribution in combating HCMV disease. Patients with defects in NK cell function exhibit extreme sensitivity to HCMV infection. We identified UL40 as the first virus gene definitively demonstrated to induce protection against NK attack, and have gone on to identify other novel NK evasion function including UL141, that contributes to immune evasion by at least three distinct pathways:

More recently we have been collaborating to use cutting edge proteomics techniques to dissect the specific ways in which HCMV influences the immune system. Through this work, we have identified an entire family of genes that act together to regulate NK cell activation through multiple mechanisms:

Not only has our work with HCMV provided information on how HCMV avoids being killed by the immune system, it has also improved our fundamental understanding of the way in which the immune system acts to recognise pathogen infected cells:

In vivo modelling of CMV infection

To examine the immune responses that control CMV infection in vivo, we utilise the murine CMV model (MCMV) of infection. Using this model system, we have investigated how soluble immune proteins called cytokines orchestrate antiviral cellular immune responses and have identified how certain immune inhibitory cytokines and other suppressive pathways suppress antiviral immunity and thus allow CMV to persist.

We now wish to understand the cellular mechanisms that regulate inflammatory versus inhibitory immune pathways. Our belief is that by understanding these complex mechanisms we will identify how, in cases of virus-induced inflammation, we can treat disease. Moreover, these studies will identify immune pathways that may be stimulated to enhance virus-induced immune responses, for example during vaccinations with viral-based vaccine vectors.

AdZ system

A recombinant Adenovirus (Ad) vector with Zero cloning steps.

The AdZ system is designed to allow you to clone a gene, shRNA or synthesized sequence directly into a replication deficient recombinant Adenovirus (Ad) vector in a single, simple, recombineering step within E.coli.

There is no conventional cloning step; the gene is simply PCR'd then recombineered directionally into the vector, and selectable markers permit the easy identification of positive colonies. Following sequencing, the vector DNA is maxiprepped and transfected into packaging cells in order to grow the recombinant Ad.

Through the use of a tet-repression system transgene expression is prevented during vector growth, thus permitting the cloning of toxic gene products. Expression of the transgene in other cell lines is constitutively on.

Transgene Insertion

  • A complete Ad5 vector is carried on a Bacterial Artificial Chromosome (BAC).
  • The DNA insert is transformed directly into cells carrying the AdZ BAC.

The insert can be:

  • Synthetic oligonucleotides (e.g. encoding shRNAs)
  • PCR product
  • Synthesized gene
  • Conventional plasmid clone (e.g. from an expression library)

Recombineering is performed:

  • The transgene replaces dual selectable markers
  • Positive clones are identified without need for screening
  • BAC DNA is purified & transfected into cells
  • AdZ recombinant grows

Protocols

Vector Maps for the AdZ vectors

The following maps are all just the expression cassettes - the remaining sequence is identical to pAdZ5-CV5 above.

Schools

School of Medicine

Next steps