HIV and Dendritic Cells
Lab head: Tony Cunnigham
Location: Centre for Virus Research, Westmead Millennium Institute
Lab members: Tony Cunningham
Funding: NHMRC / ACH2
Research approach equipment: Investigation of How HIV manipulates dendritic cells in order to fasciliate its transport to lymph nodes and transfer to T-cells
Harman AN, Lai J, Turville S, Samarajiwa S, Gray L, Marsden V, Mercier S, Jones K, Nasr N, Rustagi A, Cumming H, Donaghy H, Mak J, Gale M Jr, Churchill M, Hertzog P, Cunningham AL.
Blood. 2011 Jul 14;118(2):298-308. Epub 2011 Mar 16.
Cameron PU, Saleh S, Sallmann G, Solomon A, Wightman F, Evans VA, Boucher G, Haddad EK, Sekaly RP, Harman AN, Anderson JL, Jones KL, Mak J, Cunningham AL, Jaworowski A, Lewin SR.
Proc Natl Acad Sci U S A. 2010 Sep 28;107(39):16934-9. Epub 2010 Sep 13.
Cunningham AL, Donaghy H, Harman AN, Kim M, Turville SG.
Curr Opin Microbiol. 2010 Aug;13(4):524-9. Epub 2010 Jul 2. Review.
Harman AN, Kraus M, Bye CR, Byth K, Turville SG, Tang O, Mercier SK, Nasr N, Stern JL, Slobedman B, Driessen C, Cunningham AL.
Blood. 2009 Jul 2;114(1):85-94. Epub 2009 May 12.
Lai J, Bernhard OK, Turville SG, Harman AN, Wilkinson J, Cunningham AL.
J Biol Chem. 2009 Apr 24;284(17):11027-38. Epub 2009 Feb 17.
Donaghy H, Bosnjak L, Harman AN, Marsden V, Tyring SK, Meng TC, Cunningham AL.
J Virol. 2009 Feb;83(4):1952-61. Epub 2008 Dec 10.
Watson S, Mercier S, Bye C, Wilkinson J, Cunningham AL, Harman AN.
Virol J. 2007 Dec 3;4:130.
Cunningham AL, Harman AN, Donaghy H.
Nat Immunol. 2007 Jun;8(6):556-8. No abstract available.
- [PubMed - indexed for MEDLINE]
Harman AN, Wilkinson J, Bye CR, Bosnjak L, Stern JL, Nicholle M, Lai J, Cunningham AL.
J Immunol. 2006 Nov 15;177(10):7103-13.
The Role of Interferon Regulatory Factors in HIV Induction of Interferon in Dendritic Cells
Primary supervisor: Andrew Harman
Dendritic cells (DC) are potent antigen presenting cells that form a link between the innate and adaptive immune systems. DCs in the genital mucosa represent the first line of contact between the immune system and HIV and act to pass the virus onto CD4 T-cells, which HIV explosively replicates in. DCs therefore play a vital role in the early stages of HIV infection. When cells come into contact with a foreign antigen they secrete antiviral molecules called interferons (IFN) which help to protect them and bystander cells from viral infection by inducing the expression of hundreds of IFN stimulated genes (ISG). We have recently shown that HIV is able to block the production of IFN by dendritic cells, yet at the same time is able to induce the expression of a few ISGs. The IFN regulatory factor (IRF) family are known to play key roles in modulating both IFN induction and the expression of ISGs.
To establish the role of specific IRFs in HIV induction of ISGs in DCs.
We have recently shown that following HIV infection of DCs a small subset of ISGs is up-regulated. This ISG subset all contain a common promoter binding site for IRF1, IRF2 and IRF7 and we have gone on to demonstrate and essential role for IRF1 in the induction of these genes. We would now like to examine the roles of IRF2 and IRF7 in this process. In addition we will examine the roles of other IRFs such as IRF4 and IRF8, which are among the subset of ISGs up-regulated by HIV.
Firstly we will determine whether the proteins encoded by these genes are also up-regulated. We will then express a variety of IRFs in 293T cells to examine the independent gene profiles that these transcription factors are able to trigger. We will also over express these proteins in DCs and determine the effects on HIV replication. Finally key IRFs will be silenced in DCs and the HIV infectivity levels determined alongside the ability to induce interferon.
This project will provide a comprehensive education in isolation of dendritic cells from human blood; HIV infectivity assays, quantitative PCR and microarrays (QPCR) for determining changes in gene expression levels; silencing RNA technology; western blot and flow cytometry to determine protein expression levels.
Discipline: Infectious diseases and Immunology
Co-supervisors: Najla Nasr, Anthony Cunningham
Keywords: Dendritic cells, HIV infection, Immune response
Contact: Email Andrew Harman