HIV MOLECULAR PATHOGENESIS LABORATORY

Lab head: Pr Tony cunnigham
Location: Centre for Virus Research, Westmead Millennium Institute, Westmead, NSW, 2145

Our studies have focused on how HIV subverts the cellular environment of dendritic cells (DC) and macrophages to facilitate HIV replication and spread. In recent studies we have examined HIV uptake and replication in a variety of epithelial DCs. We have demonstrated that two-phase oh HIV transfer occurs in Langerhans cells and both phases of uptake and transfer to T cells can be blocked by soluble langerin homologues (Nasr et al., J Immunology 2014). Previously we have shown that HIV induces host DC gene expression in two phases corresponding to HIV trafficking in DCs (Harman et al., Blood 2009). Firstly we showed that de novo infection results in enhanced maturation of DCs and adherence to T cells to facilitate HIV transfer. Surprisingly microvesicles liberated after lytic infection of T cells contaminate the HIV inoculum and provide a major contribution to the maturation and adherence of these DCs. The full proteome of these vesicles including the HIV and cellular molecules responsible for the effect (nef and HSP90) has been defined (Harman et al, J Immunology 2006; Mercier et al., Plos pathogen 2013). Finally, HIV infection of both DCs and macrophages inhibits their ability to produce interferon, thus facilitating spread to adjacent cells (where interferon has its effect). Paradoxically HIV also induces interferon-stimulated genesin infected DCs and macrophages to control viral replication and perhaps lysis of these cells (Harman et al, Blood 2011, Nasr et al., Blood 212, Harman et al J Virology 2015). Thus HIV induces a persistent noncytopathic infection in these cells, ideally suited to their transfer and reservoir function. Recently we are defining the exact pathways that are responsible for these effects (Nasr et al., J Virology 2017).

A fascinating future for HIV research lies ahead as the HIV scientific community focuses on the two major aims of preventing HIV infection through vaccine/microbicide development and curing or eliminating HIV from antiretroviral-controlled chronically infected people.

Lab members: 2 post doc, 1 PhD students, 1 master student and 1 Honours student
Funding: NHMRC / ACH2
Research approach equipment: This project will provide comprehensive training in isolation and culture of monocytes from human blood; HIV infectivity assays, quantitative PCR for determining changes in gene expression levels; silencing RNA technology; QPCR western blot and flow cytometry to determine protein expression levels.
Publications:

1. Nasr N,Lai J, Botting RA, Mercier SK, Harman AN, Kim M, Turville SG, Center RJ, Domagala T, Gorry PR, Olbourne N, Cunningham AL.Inhibition of Two Temporal Phases of HIV-1 Transfer from Primary Langerhans Cells to T Cells: the Role of Langerin. J Immunol 2014

2. Mercier SK, Donaghy H,Botting R, Turville SG, Harman AN, Nasr N, Ji H, Kusebauch U, Mendoza L, Shteynberg D, Sandgren K, Simpson RJ, Moritz RL, Cunningham AL. The microvesicle component of HIV-1 inocula modulates dendritic cell infection, maturation, and enhances adhesion to and activation of T-lymphocytes. Plos Path. 2013 e1003700

3. Harman AN, Bye CR, Nasr N, Sandgren KJ, Kim M, Mercier SK, Botting RA, Lewin SR, Cunningham AL, Cameron PU. Identification of lineage relationships and novel markers of blood and skin human dendritic cells. J Immunology 2013 190:66-79

4. Harman AN, Kraus M, Bye CR, Byth K, Turville SG, Tang O, Mercier SK, Nasr N, Stern JL, Slobedman B, Driessen C, Cunningham AL. HIV-1-infected dendritic cells show 2 phases of gene expression changes, with lysosomal enzyme activity decreased during the second phase. Blood 2009 114:85-94.

5. Nasr N, Maddocks S, Turville SG, Harman AN, Woolger N, Helbig KJ, Wilkinson J, Bye CR, Wright TK, Rambukwelle D, Donaghy H, Beard MR, Cunningham AL. HIV-1 infection of human macrophages directly induces viperin which inhibits viral production Blood 2012 12:778-788.

6. Harman AN, Lai J, Turville SG, Samarajiwa S, Gray L, Marsden V, Mercier S, Jones K, Nasr N, Cumming H, Donaghy H, Mak J, Churchill M, Hertzog P, Cunningham AL. HIV infection of dendritic cells subverts the interferon induction pathway via IRF1 and inhibits type 1 interferon production. Blood 2011 118:298-308

7. Harman AN, Kraus M, Bye CR, Byth K, Turville SG, Tang O, Mercier SK, Nasr N, Stern JL, Slobedman B, Driessen C, Cunningham AL. HIV-1-infected dendritic cells show 2 phases of gene expression changes, with lysosomal enzyme activity decreased during the second phase. Blood 2009 114:85-94.

8. Gray L, Sterjovski J, Churchill M, Ellery P, Nasr N, Lewin SR, Crowe SM, Wesselingh SL, Cunningham AL, Gorry PR. Uncoupling coreceptor usage of HIV-1 from macrophage tropism reveals biological properties of CCR5-restricted HIV-1 isolates from patients with acquired immunodeficiency syndrome. Virol 2005 337:384-398

Books and reviews

1. Nasr N, Harman A, Turville S, Cunningham AL.HIV infection of dendritic cells. Methods Mol Biol. 2014 1087:221-32.

2. Harman A, Kim M,Nasr N, Cameron PU.Tissue dendritic cells as portals for HIV entry. Advances Med Virol.2013 5:319-33. C IF:7.615

3. Cunningham AL, Harman N, Nasr N. Initial HIV mucosal infection and dendritic cells. EMBO Mol Med 2013 5:658-660 C IF:7.8

4. Cunningham AL, Harman AN, Kim M,Nasr N, Lai J.Immunobiology of dendritic cells and the influence of HIV infection. Advances in Experimental Medicine and Biology 2013 762:1-44C:1 IF:1.09

5. Cunningham AL, Abendroth A, Jones C,Nasr N, Turville S.Viruses and Langerhans cells. Immunol Cell Biol. 2010 88:416-423. C:11 IF:3.66


Defining the Interaction of HIV with the Interferon System in Initial Mucosal Infection.

Primary supervisor: Najla Nasr

Background:

There is still no vaccine for HIV/AIDS and the only available treatment is combined antiretroviral therapy, which although effective at suppressing the virus in a patient, does not eliminate all reservoirs of HIV in the body and is therefore not curative.  it is clear that a much better understanding of how HIV manipulates the immune system during initial transmission is needed. Such knowledge would provide key insights into how to block the establishment of infection and also in limiting the seeding of persistent HIV cellular reservoirs. This study will look at the complex interplay between HIV and one arm of the early innate immune response to the virus - the interferon response.

AIM: To determine if HIV-infected myeloid DCs and macrophages induce the secretion of IFNα by plasmacytoid DCs and whether this affects HIV production and transfer to T cells.

This project will provide comprehensive training in the isolation of different T cell subsets and the generation of macrophages and dendritic cells from human blood; HIV infectivity assays, qPCR and  flow cytometry.


Discipline: Medical Sciences, Westmead
Co-supervisors: Anthony L Cunnigham
Keywords: Microbiology, Molecular biology, HIV infection
Contact: