The Dendritic Cell Biology & Therapeutics Group
Lab head: Professor Derek Hart
Location: ANZAC Research Institute, Concord Hospital
The Dendritic Cell Biology and Therapeutics Group at the ANZAC Research Institute is a newly established research program focused on understanding dendritic cell biology to enable the development of new ways to treat haematological diseases. The group is a translational research group with projects suitable for candidates interested in basic biology or more clinically focused work. The four main areas of focus include:
1. DC Antigen Discovery and Antibody Engineering
We have identified a number of potential novel biomarkers expressed by dendritic cells. An interested student would design and generate reagents to follow the expression of these biomarkers to discover how they may contribute to dendritic cell function.
Part of our core business is generating antibodies that can be developed for the purpose of using dendritic cells for the diagnosis and treatment of haematological diseases. In this project a student would gain the skills in antibody engineering and phage display. These techniques will be used to develop humanised antibodies and identify novel proteins that bind dendritic cells.
2. Dendritic Cell Biology – Fundamental and Applied
CD300 is a family of immune-regulatory proteins that can positively and negatively regulate dendritic cell signalling. This project would allow a student to investigate the contribution of CD300 molecules in haematological diseases.
CD302 is a DC specific molecule that plays a role in the migration of dendritic cells. A student would be involved in characterising a gene knockout model of CD302 and the effects on dendritic cells.
AHCYL-1 is another DC molecule that we have found be important in Ca2+ signalling in dendritic cells. Like the CD302 project, a student would be involved in characterising a gene knockout model.
3. Immunostimulation/Cancer Vaccination
We are developing and optimising strategies the purification and clinical use of dendritic cells to enhance immunotherapy treatment options for patients with multiple myeloma, acute myeloid leukemia and prostate cancer. The strategies being developed have the potential to be generic eventually allowing some overlap with other disease treatments.
Potential students would participate in the development of therapeutic dendritic cell vaccinations strategies and apply these to preclinical animal models of haematological malignancies. Students would gain experience in several molecular techniques including real time PCR, cloning, in vitro-transcription of mRNA tumour antigens. Additionally the student will be exposed to a variety of cellular immunological techniques including cytokine analysis, generation of tumour specific cytotoxic T cells as well as gain experience in cell isolations strategies using flow cytometry.
Our program is ultimately focused on developing novel therapies for haematological disease such as multiple myeloma and acute myeloid leukemia and understanding the balance between graft versus leukemia (GVL) and graft versus host disease (GVH) in Bone Marrow Transplantation. We have a number of projects investigating the role of dendritic cells in these conditions using patient samples.
In particular, we are developing therapeutic antibodies that can be used to deplete activated dendritic cells causing GVHD whilst retaining the dendritic cells required for GVL. Students would gain experience in flow cytometry, cellular assays and animal models and work closely with clinicians actively involved in treating these conditions. There is an opportunity to work as a student on aspects of our novel anti-CD83 therapeutic project.
Lab members: Professor Derek Hart Professor Ken Bradstock, Westmead Hospital A/Prof Georgina Clark Dr Nirupama Verma Dr Xinsheng Ju Dr Kifah Shahin, Westmead Hospital Dr Phillip Fromm, Royal Prince Alfred Hospital Dr Pablo Silveira Dr Robin Gasiorowski Dr Christian Bryant Zamil Mattar Fiona Kupresanin Ai Vu Leticia Muusers Dr Sebastien Anguille Amanda Afyouni, Executive Assistant James Zagarella, Business Manager
Wilkinson, R., Woods, K., D'Rozario, R., Prue, R., Vari, F., Hardy, M., Dong, Y., Clements, J., Hart, D., Radford, K. (2012), Human kallikrein 4 signal peptide induces cytotoxic T cell responses in healthy donors and prostate cancer patients. Cancer Immunology, Immunotherapy. 61(2), 169-179
Freeman, L., Lam, A., Petcu, E., Smith, R., Salajegheh, A., Diamond, P., Zannettino, A., Evdokiou, A., Luff, J., Wong, P., Khalil, D., Waterhouse, N., Vari, F., Rice, A., Catley, L., Hart, D., Vuckovic, S. (2011), Myeloma-induced alloreactive T cells arising in myeloma-infiltrated bones include double-positive CD8+CD4+ T cells: evidence from myeloma-bearing mouse model. Journal of Immunology. 187(8), 3987-3996.
Fromm, P., Gottlieb, D., Bradstock, K., Hart, D. (2011), Cellular therapy to treat haematological and other malignancies: progress and pitfalls. Pathology. 43(6), 605-15.
Hart, D. (2011), The delivery of effective therapeutic cancer vaccination. Asian Journal of Andrology. 13(2), 183-184.
Dean, M., Flower, R., Eisen, D., Minchinton, R., Hart, D., Vuckovic, S. (2011), Mannose-binding lectin deficiency influences innate and antigen-presenting functions of blood myeloid dendritic cells. Immunology. 132(2), 296-305.
Ding, Y., Ju, X., Azlan, M., Hart, D., Clark, G. (2011), Screening of the HLDA9 panel on peripheral blood dendritic cell populations. Immunology Letters. 134(2), 161-166.
Christensen, M., Turner, B., Sinfield, L., Kollar, K., Cullup, H., Waterhouse, N., Hart, D., Atkinson, K., Rice, A. (2010), Mesenchymal stromal cells transiently alter the inflammatory milieu post-transplant to delay graft-versus-host disease. Haematologica. 95(12), 2102-2110.
Ziegler-Heitbrock, L., Ancuta, P., Crowe, S., Dalod, M., Grau, V., Hart, D., Leenen, P., Liu, Y., MacPherson, G., Randolph, G., Scherberich, J., Schmitz, J., Shortman, K., Sozzani, S., Strobl, H., Zembala, M., Austyn, J., Lutz, M. (2010), Nomenclature of monocytes and dendritic cells in blood. Blood. 116(16), e74-e80.
Gowans, E., Roberts, S., Jones, K., Dinatale, I., Latour, P., Chua, B., Eriksson, E., Chin, R., Li, S., Wall, D., Sparrow, R., Moloney, J., Loudovaris, M., Ffrench, R., Prince, H., Hart, D., Zeng, W., Torresi, J., Brown, L., Jackson, D. (2010), A phase I clinical trial of dendritic cell immunotherapy in HCV-infected individuals. Journal of Hepatology. 53(4), 599-607.
Jongbloed, S., Kassianos, A., McDonald, K., Clark, G., Ju, X., Angel, C., Chen, C., Dunbar, P., Wadley, R., Jeet, V., Vulink, A., Hart, D., Radford, K. (2010), Human CD141+ (BDCA-3)+ dendritic cells (DCs) represent a unique myeloid DC subset that cross-presents necrotic cell antigens. Journal of Experimental Medicine. 207(6), 1247-1260.
Jones, M., Seldon, T., Smede, M., Linville, A., Chin, D., Barnard, R., Mahler, S., Munster, D., Hart, D., Gray, P., Munro, T. (2010), A method for rapid, ligation-independent reformatting of recombinant monoclonal antibodies. Journal of Immunological Methods. 354(1-2), 85-90.
Kassianos, A., Jongbloed, S., Hart, D., Radford, K. (2010), Isolation of human blood DC subtypes. Methods in Molecular Biology. 595, 45-54.
Ju, X., Clark, G., Hart, D. (2010), Review of human DC subtypes. Methods in Molecular Biology. 595, 3-20.
Targeting novel dendritic cell molecules to induce therapeutic anti-cancer T cell responses.
Primary supervisor: Derek Hart
The Dendritic Cell Biology and Therapeutics Group at the ANZAC Research Institute has forged a strong reputation in fundamental human dendritic cell research. Using its extensive clinical links and commercial collaborations, it is developing diagnostic and therapeutic antibodies for novel targets with application in clinical transplantation and the treatment of haematological and other malignancies. One promising strategy we are currently investigating using our novel targets are the use of antibodies tagged with tumour antigens that allow directed antigen delivery to dendritic cells. This approach will allow shaping of immune responses directed towards a patient’s tumour. Such immune therapies are attractive due to their high degree of specificity and the potential to target patients in states of minimal residual disease following conventional therapies.
This honours project will explore how antigen targeting to dendritic cells can be used to treat patients with haematological malignancies such as multiple myeloma and acute myeloid leukemia. This will involve characterizing how antigens delivered by antibodies are internalized by dendritic cells and the way in which these traffic to intracellular compartment using flow cytometry and confocal microscopy. The effects these targeting strategies have on inducing T cell responses to both viral and tumour antigens in both normal healthy donors and in primary samples from patients with hematological malignancies will be assessed using in vitro assays.
Students will learn how to use flow cytometry to analyze antigen uptake in primary human dendritic cells, the monitoring of T cell differentiation in response different DC targeting strategies, including the monitoring of proliferation and cytokine responses as well as measurement of T cell cytotoxicity against tumour targets. Students will also gain experience in cloning of tumour antigens, confocal microscopy, cell culture of primary human samples and molecular and cellular analysis of antigen processing. It is expected that this research will provide novel information on new targeting strategies that may have direct therapeutic implications.
Discipline: Infectious diseases and Immunology
Co-supervisors: Phillip Fromm, Georgina Clark
Keywords: Dendritic cells, T-cells, Haematological malignancy