The Sir Albert Sakzewski Virus Research Centre (SASVRC) is the Clinical Medical Virology Centre of the Faculty of Science. It also has close ties with the Health Sciences Faculty. SASVRC provides a vibrant 'biosciences milieu', and operates within the interactive research environment of the Queensland Institute of Medical Research, Royal Brisbane and Women's and Royal Children's Hospitals and the Medical School on the Herston campus.

SASVRC currently comprises approximately 35 scientists, medical doctors and students. Six research units headed by internationally acclaimed scientists conduct research in a wide range of infectious diseases (particularly viral diseases) related to human health. Research is ongoing in a range of respiratory viruses, cytomegalovirus and papillomavirus, with particular areas of excellence in viral pathogenesis, identification of new and emerging viruses, new diagnostics and therapeutics, viral immune escape mechanisms, immune response, and new viral vaccine strategies and clinical trials. Very many FOS graduates have carried out their Honours or PhD studies at SASVRC, and a good number have subsequently progressed on to high profile postdoctoral positions internationally and nationally. Grades obtained from students in recent years have been of the highest order. SASVRC offers two Honours and one PhD scholarship. The following Honours projects are available at SASVRC for the academic year 2011-2012.

HERPESVIRUS MOLECULAR PATHOGENESIS UNIT

Unit Head      Dr. Nick Davis-Poynter. Tel: 3636-8138.  Email: n.davispoynter@uq.edu.au

The cytomegaloviruses (CMVs) are ubiquitous herpesviruses that establish persistent infections in the host.  Whilst they cause mild symptoms in healthy individuals, they a major cause of morbidity and mortality amongst those whose immunity is immature (the neonate) or immunocompromised (AIDS patients and transplant recipients).  Current antiviral therapies that target the replicative machinery of CMV suffer from toxic effects and the emergence of drug-resistant strains.  Thus, there is a need to identify alternative strategies to inhibit the replication and dissemination of CMV in the susceptible host.
The ability of CMVs to persist in their hosts is due in part to an orchestrated expression of virus proteins that evade or sabotage the host immune response.  Our group is determining the contribution of these proteins to the CMV lifecycle and to evaluate their potential for novel antiviral therapies.  We utilise a mouse model for human CMV, as this allows us to study the function of these immune-evasive proteins from the cellular to the whole animal level.  Our current studies focus on homologues of chemokine receptors that are encoded by CMV (viral chemokine receptors or vCKR).   Cellular CKR are cell surface proteins that signal via intracellular G proteins and invoke a cascade of intracellular signalling pathways; they are found on most leukocytes and co-ordinate their migration (chemotaxis) throughout the body.  Given that a major part of the CMV lifecycle occurs in leukocytes, our working hypothesis is that the vCKR exploit or interfere with normal chemokine signalling pathways to promote virus dissemination and/or establish and intracellular environment conducive to virus replication. Recent published and unpublished data from our laboratory and those of our collaborators support this hypothesis.

The projects offered will investigate the vCKR of mouse CMV (termed M33 and M78).  Disruption of either of these genes results in profound attenuation in vivo, suggesting they may be novel targets for antiviral drugs. 

Project One. 

Functional studies of M33 signalling through site-directed mutagenesis.  To date, studies from our laboratory have shown that M33 is critical for virus dissemination and that the ability of M33 to engage in G protein-signalling is important.  Whilst we have a broad view of the regions of the M33 protein that are critical for signalling, more information is required to identify the “hot-spots” of M33 that is important for its function.  This project will utilise site-directed mutagenesis to further define the M33 motifs that determine its signalling profile in vitro and its function in vivo.  The project will involve:
(i) The design and construction of specific mutations in M33:
(ii) Identification of the effects of mutations on M33 signalling in vitro;
(iii) The construction of mouse CMV recombinants expressing the desired M33 mutations
(iv) Investigation of the effects of introduced mutations on well-established virological in vivo and in vitro assays.
This project offers the opportunity for the student to follow the effects of M33 mutation in vitro and in vivo.  The project will encompass a wide range of molecular, cellular and virological techniques.

Project Two. 

The role of intracellular trafficking of M78 on virus replication.  Like many viral chemokine receptors, M78 undergoes constitutive endocytosis and recycling to the cell membrane. For most chemokine receptors, endocytosis requires the presence of particular motifs within the carboxy tail of the receptor.  Preliminary unpublished studies have shown that similar to cellular chemokine receptors, the truncation of the M78 carboxy tail modulates rates of M78 endocytosis and significantly, this modification affects the ability of the virus to replicate in vitro.  This project will investigate the correlation between the rate of endocytosis of M78 and its contribution to virus replication.  The project will involve:
(i) The design and construction of specific carboxy-tail mutations of M78;
(ii) Identification of the effects of the mutations on M78 endocytosis in transfected cells by immunofluorescent histochemistry;
(iii) The construction of mouse CMV recombinants expressing the desired M78 mutations;
(iv) The analysis of growth of M78 mutants in defined cell lines in vitro.
(v)  If time permits, the effect of M78 mutation on M78 morphogenesis will be assessed by electron microscopy.
As with project one, this project offers the opportunity for the student to follow the effects of mutagenesis from a molecular to the cellular level. The project encompasses molecular, histological and virological techniques.

 

Recent relevant publications
Case R, Sharp E, Benned-Jensen T, Rosenkilde MM, Davis-Poynter N, Farrell HE. (2008)
Functional analysis of the murine cytomegalovirus chemokine receptor homologue M33: ablation of constitutive signalling is associated with an attenuated phenotype in vivo. J Virol. 82:1884-1898.

Cardin RD, Schaefer GC, Allen JR, Davis-Poynter NJ & Farrell HE. (2009).
The M33 chemokine receptor homolog of murine cytomegalovirus exhibits a differential tissue-specific role during in vivo replication and latency.  J. Virol.83:7590-7601.

Sharp EL, Davis-Poynter NJ & Farrell HE (2009).
Analysis of the subcellular trafficking properties of murine cytomegalovirus (MCMV) M78, a 7 transmembrane receptor homologue. J. Gen. Virol. 90:59-68.

Farrell HE, Abraham AM, Cardin RD, Sparre-Ilrich AH, Rosenkilde MM, Spiess K, Jensen TH, Kledal TN & Davis-Poynter N (2011).
Partial functional complementation between human and mouse cytomegalovirus chemokine receptor homologues  J. Virol 85:6091-6095.

 

RESPIRATORY VIRUS RESEARCH UNIT

Unit Head      Dr. Kirsten Spann. Tel: 3636-8718  Email:  k.spann@uq.edu.au

The long-term vision of RVRU is to develop antiviral treatments for paediatric respiratory viruses, and to contribute to the world-wide effort to develop effective vaccines for hRSV and hMPV . The unit will focus on Respiratory Syncytial Virus (RSV) and human Metapneumovirus (hMPV). RSV is the principal cause of severe lower respiratory tract disease in infants and young children, and can also cause serious disease in immunocompromised adults and the elderly. Clinical manifestations vary from rhinitis and otitis media, to bronchiolitis and pneumonia. The global annual RSV infection rate is estimated to be 64 million, with approximately 160,000 cases resulting in death. The hospitalization burden due to RSV infection in Australia is considerable. Various research efforts are underway to develop vaccines to combat RSV. However none have been licensed. MPV is a significant cause of both upper and lower respiratory disease in infants and children. The clinical symptoms are similar to those of RSV and by age 5 virtually all children have been exposed to hMPV. Both RSV and hMPV have been associated with exacerbated asthma. RVRU is also involved in elucidating the mechanisms by which viruses trigger asthma exacerbation. 

Projects

• Functional domains of RSV non-structural proteins

Human Respiratory Syncytial Virus (hRSV) regulates both type I interferon -dependent and -independent antiviral responses via two non-structural proteins (NS1 and NS2).  For this reason, these proteins are viable targets for antiviral therapies and improved vaccine candidates. However prior to designing antiviral agents and engineering modified live vaccine candidates, the functional regions of the proteins need to be identified. By developing live mutated RSVs and expression of individual proteins, we are identifying the regions of NS1 and NS2 involved in antagonising the antiviral response of host cells. This project would involve the construction of live recombinant viruses containing specific mutations and analysis of how changes in NS1 and NS2 affect their function, particularly as antiviral response antagonists.  These analyses required would include cell culture, viral replication, protein expression, qPCR and immunological assays.

• The differences in the innate immune response of respiratory epithelial cells from asthmatic and non-asthmatic children

Respiratory viruses first invade the upper airway and then may progress to infect the lower airway. Increasing data suggests that asthmatics may be less able to prevent this spread, related to a decrease in the anti-viral innate immune response in their respiratory epithelial cells. We do not know if this defect is equally present in the upper and lower airway cells or whether this contributes to acute viral-associated exacerbations of asthma.  We aim to identify differences in the innate immune response of airway epithelial cells from asthmatic and non-asthmatic children to viral infection.

In this project we will collect nasal (upper) and tracheal (lower) epithelial cells from atopic asthmatic, atopic non-asthmatic, non-atopic asthmatic and non-atopic non-asthmatic children from 2-10 years admitted to the RCH for elective surgery. These cells will be grown in culture and infected with 2 viruses which commonly trigger asthma exacerbations; respiratory syncytial virus (RSV), and metapneumovirus (MPV). The innate immune responses to these viruses and viral replication will be quantified. Specifically, the induction of type I (a/b) and type III (l) interferon and pro-inflammatory cytokines will be measured using quantitative (q)RT-PCR for gene transcription and Bio-PLEX and ELISA assays for protein secretion. Antiviral protein expression will be measured by western blot analysis. Viral replication will be measured by titrating virus shed from infected cells over 5 days. The project will involve cell culture, and viral propagation and infection, in addition to immunological assays.

 

QUEENSLAND PAEDIATRIC INFECTIOUS DISEASES (QPID) LABORATORY

A Prof Ian M Mackay. Tel: 3636-1619  Email:  ian.mackay@uq.edu.au

The Mackay group in the Queensland paediatric Infectious Diseases (QPID) Laboratory seeks to discover, identify and characterize respiratory viruses from ill children. Improving basic knowledge and understanding the mechanisms which underpin the clinical impact of infection are the first steps toward targeted research for each virus. Respiratory virus discovery and characterization precedes the development of specific diagnostics and epidemiology studies, it informs patient management and can prioritize antiviral and vaccine research. This work will be supported by grants from the ARC and the Queensland Children’s Medical Research Institute (QCMRI).

Projects

1. Human rhinoviruses and enteroviruses in asthma: identification, molecular analysis and clinical impact.

Respiratory picornaviruses, especially human rhinoviruses (HRVs), are a common environmental factor linked to exacerbations in those with asthma but little is known of the role of the different genetic clades in illness or in triggering early events in our interferon-based antiviral response to their presence. This project will access clinically well-defined specimens from hospitalised and non-hospitalised children and analyse them for the presence and genotype of human rhinoviruses (HRVs) and enterovirus (EVs) present. Select virus types will be cultured and additional genome deduction will be conducted on viral types of interest. A pilot study will be conducted to examine the feasibility of using PCR-based analyses to correlate the presence of HRVs and EVs and the RNA levels of the immune markers PKR, OAS, MxA, RIG-I, IRF3, viperin and CXCL10.

1. Genome deduction and epidemiology of a recently identified picornavirus

The past few years have seen the discovery of many previously unknown picornavirus types, species and genera from ill human subjects. Very little is known of their genetic diversity, the conservation of their genomic features within a clade or the seasonality of each virus in the respiratory tract. This project will focus on Saffold viruses (SAFVs), members of a species of the same name within the genus Cardiovirus, which we have identified in the respiratory tract of ill Queensland children. Existing SAFV-positive samples will serve as the source for genome sequencing and culture while additional SAFVs will be sought during PCR-based molecular epidemiology investigations. Extensive oligonucleotide design will be used to gain “footholds” across the ~8kb genome sequence and to “walk” along the SAFV sequence. The complete polyprotein sequence will be compared and contrasted to SAFVs and other picornavirus sequences from around the world. Additionally, an estimate of the prevalence and seasonal spread of SAFV in Queensland’s hospital-based populations during 2011 will be determined.