Mucosal Immunology and Sexual Transmission of HIV

Lab head: Andrew Harman
Location: Centre for Virus Research, The Westmead Institute for Medical Research

Lab members: Andrew Harman Kirstie Bertram Heeva Baharlou Hafsa Rana Jake Rhodes
Funding: NHMRC
Research approach equipment: Our group has acquired access to all the tissues that make up the human anogenital tracts (labia, vagina, cervix, foreskin, glans penis, anus and rectum). We use these tissue to define the immune environment of these tissue and examine their role in sexual transmission of HIV.

HIV infection of dendritic cells subverts the IFN induction pathway via IRF-1 and inhibits type 1 IFN production.

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.

Establishment of HIV-1 latency in resting CD4+ T cells depends on chemokine-induced changes in the actin cytoskeleton.

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.

Manipulation of dendritic cell function by viruses.

Cunningham AL, Donaghy H, Harman AN, Kim M, Turville SG.

Curr Opin Microbiol. 2010 Aug;13(4):524-9. Epub 2010 Jul 2. Review.

HIV-1-infected dendritic cells show 2 phases of gene expression changes, with lysosomal enzyme activity decreased during the second phase.

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.

Oligomerization of the macrophage mannose receptor enhances gp120-mediated binding of HIV-1.

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.

Role for plasmacytoid dendritic cells in the immune control of recurrent human herpes simplex virus infection.

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.

Determination of suitable housekeeping genes for normalisation of quantitative real time PCR analysis of cells infected with human immunodeficiency virus and herpes viruses.

Watson S, Mercier S, Bye C, Wilkinson J, Cunningham AL, Harman AN.

Virol J. 2007 Dec 3;4:130.

DC-SIGN 'AIDS' HIV immune evasion and infection.

Cunningham AL, Harman AN, Donaghy H.

Nat Immunol. 2007 Jun;8(6):556-8. No abstract available.

[PubMed - indexed for MEDLINE]

HIV induces maturation of monocyte-derived dendritic cells and Langerhans cells.

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.

Exploiting skin dendritic cells in vaccination

Primary supervisor: Andrew Harman

Research Background Most vaccines today are injected into the muscle. The muscle however, is not well populated with immune cells and may therefore be a suboptimal site for vaccine delivery. The skin is now being considered as an ideal target for vaccination due to its dense network of immune cells, including dendritic cells (DCs). DCs are essential for the stimulation of protective immunity and understanding their role in vaccination may allow us to enhance and tailor immune responses to elicit more robust protection against unconquered infections such as HIV. An in vivo model for skin vaccination would allow testing of different vaccine delivery devices such as microneedle patches, and the examination of early immune events after vaccination, such as cell recruitment, activation, transport of the vaccine to the lymph nodes and stimulation of specific T cell response. Since the immune system, including the DC subsets, appear largely comparable in non-human primates (NHPs) and humans (unlike mice), NHPs offer a much more powerful model for how vaccines work in humans and would yield data that can be translated into optimising future vaccine formulations and delivery. Very little is documented about macaque skin and this project will focus on the development of such a model. Research Aims and Plan In order to develop an in vivo model in NHPs for skin vaccination, the aims of this project are to: 1. Measure the thickness of macaque skin layers and compare with human skin ¿ critical for the design of vaccine delivery devices. 2. Examine the phenotype and distribution of immune cells, particularly different DC subsets, through the various layers of macaque skin. 3. Assess the delivery and uptake of fluorescently labelled antigen via microneedles in macaque skin. The main techniques employed in this project will be cryosectioning and immunofluorescent staining of skin for microscopy using state of the art confocal and deconvolution microscopes. We will also conduct quantitative computerised image analysis. Dissociation of DCs from skin and multicolour flow cytometry will also be used for phenotypic and functional analyses if sample size permits. We receive macaque skin through local and overseas collaborations and discarded human skin from local plastic surgeons. Ultimately, information from an NHP model has the potential to reveal specialised functional traits of certain DC subsets that may be exploited to formulate more potent vaccines specifically for the skin.

Discipline: Infectious diseases and Immunology
Co-supervisors: Kerrie Sandgren, Anthony Cunningham
Keywords: Cell biology, Immunology, HIV infection