Dendritic Cell Research

Lab head: Professor Derek Hart
Location: ANZAC Research Institute, Concord Hospital

The Dendritic Cell Research Group at the ANZAC Research Institute is a recently 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 for 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.

4.     Immunosuppression/Anti-inflammatory

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 (GVHD) 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.

Website: http://www.dcresearch.org.au
Lab members: Professor Derek Hart Professor Ken Bradstock, Westmead Hospital A/Prof Georgina Clark Dr Xinsheng Ju Dr Phillip Fromm Dr Pablo Silveira Dr Ahmed Mekkawy Dr Jen Hsu Dr Robin Gasiorowski Dr Christian Bryant Ms Fiona Kupresanin Ms Kate Chen Mr Kevin Lo Mr Michael Papadimitrious Dr Blake Hsu Dr Cindy Li Dr Ben Kong Dr Sarah Sutherland
Funding: Using Immunology to Cure Patients with Blood Cancers; Enabling Diagnostic and Therapeutic Antibodies for Haematological and other Malignancies; mRNA Prostate Cancer Antigen Loaded Blood Dendritic Cell Therapy
Publications:

  1. Joshua D, Suen H, Brown R, Bryant C, Ho PJ, Hart D, Gibson J: The T cell in Myeloma. Clinical Lymphoma Myeloma and Leukemia 2016. IN PRESS  doi: 10.1016/j.clml.2016.08.003
  2. Suen H, Brown R, Yang S, Bryant C, Ho PJ, Weatherburn C, Woodland N, Nassif N, Barbaro P, Hart D, Gibson J, Joshua D: Multiple myeloma causes clonal T-cell immunosenescence: identification of potential novel targets for promoting tumour immunity and implications for checkpoint blockade. Leukemia 2016:1716-24.
  3. Seldon TA, Pryor R, Palkova A, Jones ML, Verma ND, Findova M, Braet K, Sheng Y, Fan Y, Zhou EY, Marks JD, Munro T, Mahler SM, Barnard RT, Fromm PD, Silveira PA, Elgundi Z, Ju X, Clark GJ, Bradstock KF, Munster DJ, Hart DN: Immunosuppressive human anti-CD83 monoclonal antibody depletion of activated dendritic cells in transplantation. Leukemia 2016, 30:692-700.
  4. Lo TH, Silveira PA, Fromm PD, Verma ND, Vu PA, Kupresanin F, Adam R, Kato M, Cogger VC, Clark GJ, Hart DN: Characterization of the Expression and Function of the C-Type Lectin Receptor CD302 in Mice and Humans Reveals a Role in Dendritic Cell Migration. J Immunol 2016, 197:885-98.
  5. Fromm PD, Papadimitrious MS, Hsu JL, Van Kooten Losio N, Verma ND, Lo TH, Silveira PA, Bryant CE, Turtle CJ, Prue RL, Vukovic P, Munster DJ, Nagasaki T, Barnard RT, Mahler SM, Anguille SA, Berneman Z, Horvath LG, Bradstock KF, Joshua DE, Clark GJ, Hart DN: CMRF-56(+) blood dendritic cells loaded with mRNA induce effective antigen-specific cytotoxic T-lymphocyte responses. Oncoimmunology 2016, 5:e1168555.
  6. Fromm PD, Kupresanin F, Brooks AE, Dunbar PR, Haniffa M, Hart DN, Clark GJ: A multi-laboratory comparison of blood dendritic cell populations. Clin Transl Immunology 2016, 5:e68.
  7. Clark GJ, Kupresanin F, Fromm PD, Ju X, Muusers L, Silveira PA, Elgundi Z, Gasiorowski RE, Papadimitrious MS, Bryant C, Lee KM, Clarke CJ, Young JW, Chan A, Harman A, Botting R, Cabezon R, Benitez-Ribas D, Brooks AE, Dunbar PR, Hart DN: New insights into the phenotype of human dendritic cell populations. Clin Transl Immunology 2016, 5:e61.
  8. Clark G, Stockinger H, Balderas R, van Zelm MC, Zola H, Hart D, Engel P: Nomenclature of CD molecules from the Tenth Human Leucocyte Differentiation Antigen Workshop. Clin Transl Immunology 2016, 5:e57.
  9. Bryant C, Fromm PD, Kupresanin F, Clark G, Lee K, Clarke C, Silveira PA, Suen H, Brown R, Newman E, Cunningham I, Ho PJ, Gibson J, Bradstock K, Joshua D, Hart D: A CD2 high-expressing stress-resistant human plasmacytoid dendritic-cell subset. Immunol Cell Biol 2016, 94:447-57.
  10. Prue RL, Vari F, Radford KJ, Tong H, Hardy MY, D'Rozario R, Waterhouse NJ, Rossetti T, Coleman R, Tracey C, Goossen H, Gounder V, Crosbie G, Hancock S, Diaz-Guilas S, Mainwaring P, Swindle P, Hart DN: A Phase I Clinical Trial of CD1c (BDCA-1)+ Dendritic Cells Pulsed With HLA-A*0201 Peptides for Immunotherapy of Metastatic Hormone Refractory Prostate Cancer. J Immunother 2015, 38:71-6.
  11. McAuliffe S, Brown R, Catalano A, Ho PJ, Nassif N, Woodland N, Hart D, Weatherburn C, Yang S, Suen H, Paul C, Joshua D, Gibson J: Using digital polymerase chain reaction to detect minimal residual disease in myeloma by identifying FGFR3 up-regulation. Leukemia & Lymphoma 2015:1-3.
  12. Anguille S, Smits EL, Bryant C, Van Acker HH, Goossens H, Lion E, Fromm PD, Hart DN, Van Tendeloo VF, Berneman ZN: Dendritic Cells as Pharmacological Tools for Cancer Immunotherapy. Pharmacol Rev 2015, 67:731-53.
  13. Gasiorowski RE, Clark GJ, Bradstock K, Hart DN: Antibody therapy for acute myeloid leukaemia. Br J Haematol 2014, 164:481-95.
  14. Favaloro J, Liyadipitiya T, Brown R, Yang S, Suen H, Woodland N, Nassif N, Hart D, Fromm P, Weatherburn C, Gibson J, Ho PJ, Joshua D: Myeloid derived suppressor cells are numerically, functionally and phenotypically different in patients with multiple myeloma. Leuk Lymphoma 2014, 55:2893-900.
  15. Favaloro J, Brown R, Aklilu E, Yang S, Suen H, Hart D, Fromm P, Gibson J, Khoo L, Ho PJ, Joshua D: Myeloma skews regulatory T and pro-inflammatory T helper 17 cell balance in favor of a suppressive state. Leuk Lymphoma 2014, 55:1090-8.
  16. Brown R, Yang S, Weatherburn C, Gibson J, Ho PJ, Suen H, Hart D, Joshua D: Phospho-flow detection of constitutive and cytokine-induced pSTAT3/5, pAKT and pERK expression highlights novel prognostic biomarkers for patients with multiple myeloma. Leukemia 2014.
  17. Shahin K, Sartor M, Hart DN, Bradstock KF: Alterations in chemokine receptor CCR5 expression on blood dendritic cells correlate with acute graft-versus-host disease. Transplantation 2013, 96:753-62.
  18. Hardy MY, Vari F, Rossetti T, Hart DN, Prue RL: A flow cytometry based assay for the enumeration of regulatory T cells in whole blood. J Immunol Methods 2013, 390:121-6.
  19. Gasiorowski RE, Ju X, Hart DN, Clark GJ: CD300 molecule regulation of human dendritic cell functions. Immunol Lett 2013, 149:93-100.
  20. Bryant C, Suen H, Brown R, Yang S, Favaloro J, Aklilu E, Gibson J, Ho PJ, Iland H, Fromm P, Woodland N, Nassif N, Hart D, Joshua DE: Long-term survival in multiple myeloma is associated with a distinct immunological profile, which includes proliferative cytotoxic T-cell clones and a favourable Treg/Th17 balance. Blood Cancer J 2013, 3.
  21. Wilkinson R, Woods K, D'Rozario R, Prue R, Vari F, Hardy MY, Dong Y, Clements JA, Hart DN, Radford KJ: Human kallikrein 4 signal peptide induces cytotoxic T cell responses in healthy donors and prostate cancer patients. Cancer Immunol Immunother 2012, 61:169-79.
  22. Li S, Roberts S, Plebanski M, Gouillou M, Spelman T, Latour P, Jackson D, Brown L, Sparrow RL, Prince HM, Hart D, Loveland BE, Gowans EJ: Induction of multi-functional T cells in a phase I clinical trial of dendritic cell immunotherapy in hepatitis C virus infected individuals. PloS one 2012, 7:e39368.
  23. Kassianos AJ, Hardy MY, Ju X, Vijayan D, Ding Y, Vulink AJ, McDonald KJ, Jongbloed SL, Wadley RB, Wells C, Hart DN, Radford KJ: Human CD1c (BDCA-1)+ myeloid dendritic cells secrete IL-10 and display an immuno-regulatory phenotype and function in response to Escherichia coli. Eur J Immunol 2012, 42:1512-22.
  24. Brown R, Kabani K, Favaloro J, Yang SH, Ho PJ, Gibson J, Fromm P, Suen H, Woodland N, Nassif N, Hart D, Joshua D: CD86(+) or HLA-G(+) can be transferred via trogocytosis from myeloma cells to T cells and are associated with poor prognosis. Blood 2012, 120:2055-63.
  25. Hart D: The delivery of effective therapeutic cancer vaccination. Asian J Androl 2011, 13:183-4.
  26. Fromm PD, Gottlieb D, Bradstock KF, Hart DN: Cellular therapy to treat haematological and other malignancies: progress and pitfalls. Pathology 2011, 43:605-15.
  27. Freeman LM, Lam A, Petcu E, Smith R, Salajegheh A, Diamond P, Zannettino A, Evdokiou A, Luff J, Wong PF, Khalil D, Waterhouse N, Vari F, Rice AM, Catley L, Hart DN, Vuckovic S: Myeloma-induced alloreactive T cells arising in myeloma-infiltrated bones include double-positive CD8+CD4+ T cells: evidence from myeloma-bearing mouse model. J Immunol 2011, 187:3987-96.
  28. Ding Y, Ju X, Azlan M, Hart DNJ, Clark GJ: Screening of the HLDA9 panel on peripheral blood dendritic cell populations. Immunology Letters 2011, 134:161-6.
  29. Dean MM, Flower RL, Eisen DP, Minchinton RM, Hart DN, Vuckovic S: Mannose-binding lectin deficiency influences innate and antigen-presenting functions of blood myeloid dendritic cells. Immunology 2011, 132:296-305.
  30. Ziegler-Heitbrock L, Ancuta P, Crowe S, Dalod M, Grau V, Hart DN, Leenen PJ, Liu YJ, MacPherson G, Randolph GJ, Scherberich J, Schmitz J, Shortman K, Sozzani S, Strobl H, Zembala M, Austyn JM, Lutz MB: Nomenclature of monocytes and dendritic cells in blood. Blood 2010, 116:e74-80.
  31. Mount AM, Masson F, Kupresanin F, Smith CM, May JS, van Rooijen N, Stevenson PG, Belz GT: Interference with dendritic cell populations limits early antigen presentation in chronic gamma-herpesvirus-68 infection. J Immunol 2010, 185:3669-76.
  32. Kassianos AJ, Jongbloed SL, Hart DN, Radford KJ: Isolation of human blood DC subtypes. Methods Mol Biol 2010, 595:45-54.
  33. Ju X, Clark G, Hart DN: Review of human DC subtypes. Methods Mol Biol 2010, 595:3-20.
  34. Jongbloed SL, Kassianos AJ, McDonald KJ, Clark GJ, Ju XS, Angel CE, Chen CJJ, Dunbar PR, Wadley RB, Jeet V, Vulink AJE, Hart DNJ, Radford KJ: Human CD141(+) (BDCA-3)(+) dendritic cells (DCs) represent a unique myeloid DC subset that cross-presents necrotic cell antigens. Journal of Experimental Medicine 2010, 207:1247-60.
  35. Jones ML, Seldon T, Smede M, Linville A, Chin DY, Barnard R, Mahler SM, Munster D, Hart D, Gray PP, Munro TP: A method for rapid, ligation-independent reformatting of recombinant monoclonal antibodies. J Immunol Methods 2010, 354:85-90.
  36. Gowans EJ, Roberts S, Jones K, Dinatale I, Latour PA, Chua B, Eriksson EM, Chin R, Li S, Wall DM, Sparrow RL, Moloney J, Loudovaris M, Ffrench R, Prince HM, Hart D, Zeng W, Torresi J, Brown LE, Jackson DC: A phase I clinical trial of dendritic cell immunotherapy in HCV-infected individuals. J Hepatol 2010, 53:599-607.
  37. Christensen ME, Turner BE, Sinfield LJ, Kollar K, Cullup H, Waterhouse NJ, Hart DN, Atkinson K, Rice AM: Mesenchymal stromal cells transiently alter the inflammatory milieu post-transplant to delay graft-versus-host disease. Haematologica 2010, 95:2102-10.
  38. Brown RD, Kabani K, Yang SH, Aklilu E, Ho PJ, Gibson J, Joshua DE: HLA-G and CD86 Expression on Malignant Plasma Cells Convey a Poor Prognosis and Immunoregulate T Cells In Patients With Myeloma. Blood 2010, 116:432.

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
Contact: