Malaghan Institute of Medical Research

Research Projects

Mechanisms of induction of CD8+ T cells by dendritic cells

Contributors: Taryn Osmond, Dr Troels Petersen, Dr Ian Hermans

Researchers at the Malaghan Institute and the Wellington Blood and Cancer Centre, have undertaken extensive pre-clinical and clinical analyses of vaccines based on the injection of dendritic cells that have been loaded with tumour derived material as a source of antigens. Dendritic cells belong to a subset of cells called antigen-presenting cells that take up and process molecules from the extracellular environment. The dendritic cells then present this information to other cells of the immune system, such as cytotoxic (CD8+) T cells, in a manner that stimulates immunity.

Recent studies have identified a specific subset of langerin-expressing dendritic cells in the spleen as critical for the induction of cytotoxic T cell activity. Using a mouse strain in which langerin-expressing dendritic cells can be depleted, PhD student Taryn Osmond is examining the in vivo function of these cells in more detail. The information generated from her research will help define how best to exploit the activity of different dendritic cell populations to optimise vaccination strategies for cancer therapy.

This research is supported by: the Cancer Society of New Zealand, the Health Research Council of New Zealand and The Royal Society of New Zealand Marsden Fund.

 

Using dendritic cell vaccines in the clinic

Contributors: Mr Martin Hunn, Evelyn Bauer, Catherine Wood, Dr Ian Hermans, collaborating with Dr David Hamilton, Marina Dzhelali (CCDHB) and Prof Michael Findlay (CTNZ)

In late 2008 the Malaghan Institute initiated a Phase I clinical trial in collaboration with the Capital & Coast District Health Board to test the feasibility and safety of using dendritic cell vaccines in combination with temozolomide chemotherapy for the treatment of patients with recurrent glioblastoma multiforme, a highly aggressive brain tumour with a 100% fatality rate. This trial has recently been completed, the outcomes from which are currently being analysed and prepared for publication.

Personalised vaccines, such as those being used in the glioblastoma multiforme clinical trial, are very intensive to produce but they do offer a broad immunity that recognises the unique features of an individual patient's tumour tissue. Complementing the clinical trials is an extensive basic immunology research programme involving several of the Institute's research groups, aimed at understanding anti-tumour immune responses and how they can be more effectively elicited with vaccines.

The role of langerin+CD8+ dendritic cells in generating T cell mediated anti-tumour immune responses

Contributors: John Gibbons, Dr Ian Hermans, Dr Troels Petersen

While it has long been recognised that dendritic cells are particularly potent immunostimulatory cells, and their role in generating T cell mediated immune responses is critical, it is also evident that many discrete dendritic cell subsets exist. Recent studies by the Vaccine Research Group have shown that cellular material from injected dendritic cells is acquired by lymphoid resident dendritic cells. In particular it is the langerin+ CD8+ dendritic cells that appear to play a pivotal role in inducing protective CD8+ T cell immune responses to tumours in the blood.

The targeting of vaccine antigens and adjuvant to this dendritic cell subpopulation is therefore an attractive approach for improved dendritic cell immunisation strategies. Considerable effort is now being directed towards understanding the basic biology of langerin+ CD8+ dendritic cells, as well as their role in initiating and maintaining immune responses in blood-borne tumour disease models.

Small molecules for cancer therapy

Contributors: Dr Ben Mulchin, Emma Dangerfield, Janice Cheng, Stephanie Chee, Dr James Baty, Dr Jacquie Harper, Prof Mike Berridge, Dr Ian Hermans, Dr Mattie Timmer, Dr Bridget Stocker

In a typical cancer immunotherapy programme, tumour cells are extracted from a patient and used to prepare a vaccine designed to stimulate an antitumour immune response. The addition of certain glycolipids, acting as adjuvants, boosts the immune response in favour of enhanced anti-tumour activity. While the tumour-derived peptide effectively acts as the 'ignition' and turns the immune response 'on', the glycolipid acts as the 'throttle' and controls the intensity of the immune response.

The Institute's Immunoglycomics Group, led by Dr Bridget Stocker and Dr Mattie Timmer (VUW) are currently synthesising a variety of novel glycolipids to enhance immunity to tumours. They have also generated fluorescently labelled glycolipids to investigate where they move in the body.

In addition, the Immunoglycomics group has been working towards the development of novel quinolinequinones as cancer drugs. Their efforts have proven fruitful with the identification of several lead compounds that show selective anticancer activities.

Improving vaccines with compounds that stimulate NKT cells

Contributors: Dr Troels Petersen, Dianne Sika- Paotonu, Deborah Knight, Sara McKee, Dr Bridget Stocker, Dr Ian Hermans, collaborating with Dr Gavin Painter (IRL, Wellington), Dr Sarah Young & Dr Margaret Baird (University of Otago) and Dr Gill Webster (Innate Therapeutics Ltd, Auckland)

Dr Ian Hermans' Vaccine Research Group has shown that they can generate very strong antitumour immune responses when the dendritic cell vaccine is used in combination with the glycolipid α-galactosylceramide (α-GalCer), which stimulates potent anti-tumour immune responses through activation of Natural Killer T (NKT) cells. These results complement their earlier discovery that certain combinations of Toll-like receptor ligands improve dendritic cell activity and will be taken into consideration when formulating clinical vaccine protocols.

Dr Hermans is currently working with Dr Gavin Painter from Industrial Research Limited to determine if they can identify novel glycolipids from bacteria with NKT stimulating activity.

The Vaccine Research Group is also investigating the effects of incorporating NKT cell ligands into vaccines based on virus-like particles in collaboration with Dr Sarah Young, and in bacterial nanoparticles with Dr Gill Webster.

Isolation and characterisation of GBM cancer stem cells

Contributors: Kate Broadley, Kathryn Farrand, Mr Martin Hunn, Dr Patries Herst, Dr Ian Hermans, Dr Melanie McConnell

Cancer cells grow under conditions of metabolic and energetic stress. During chemotherapy and radiotherapy treatments of cancer patients, these cells are subjected to further stress, yet some are able to survive and go on to cause relapse and metastasis. This is thought to be due to the presence of cancer stem cells, which are drug and radiation resistant.

Dr Melanie McConnell's Cell Survival Group has established various methodologies to allow identification of cancer stem cells and to characterise their function. They are using different cancer models to study the different properties of cancer stem cells – self-renewal and resistance to radiation and chemotherapy using glioblastoma multiforme (GBM) cells; drug resistance in melanoma cells; and metastasis using breast cancer cells.

They have used stem cell culture, stem cell gene markers and drug resistance to identify a population of cells in GBM tumours that are highly tumourigenic and potentially immune suppressive. Neurosurgeon and Malaghan Clinical Research Fellow Mr Martin Hunn has shown that these cells can be recognised by the immune system and protect against tumour development.

Using dendritic cell immunisation to sensitise malignant glioma to chemotherapy

Contributors: Mr Martin Hunn, Kathryn Farrand, Kate Broadley, Dr Melanie McConnell, Dr Ian Hermans

One of the long term aims of the Vaccine Research Group is to develop effective and safe immunotherapies for high grade glioma (brain cancer). Complementing the phase I GBM clinical trial is a basic research programme involving Mr Martin Hunn that is investigating the possibility of directing anti-tumour immune responses specifically against GBM.

Current work is focused on developing a murine model of glioma in which to assess the efficacy and safety of vaccination strategies directed at drug-resistant tumour stem cells, to support the translation of this novel immunotherapy into clinical trial. Mr Hunn has shown that the GBM cells can grow as detached 'neurospheres' in appropriate culture media and that these spheres show increased expression of stem cell-related genes and initiate tumours earlier compared to parental tumour cells.

Mr Hunn will also undertake an in vitro assessment of the immune responses of patients with recurrent GBM who are being treated with the dendritic cell vaccine in the phase I clinical trial.

Activating the immune system against childhood brain cancers

Contributors: Carole Grasso, Mr Martin Hunn, Dr Ian Hermans, Prof Mike Berridge

The overall goal of this research project, supported by the Child Health Research Foundation, is to apply the Institute's broad knowledge in dendritic cell immunotherapy to the treatment of childhood brain cancers, in particular the brain metastasis associated with childhood leukaemia.

To address this goal, orthotopic models of brain cancer have been established in mice including WEHI-3 myelomonocytic leukaemia, GL261 glioblastoma and EL4 thymoma. In these models GL261 grow as clearly marginated brain tumours when injected into the cerebral cortex whereas EL4 and WEHI-3 grow as diffuse tumours with poorly defined margins. Significant progress has been made investigating dendritic cell immunotherapy with GL261 tumours grown subcutaneously, but the time-consuming nature of the orthotopic brain tumour model has restricted the number of experiments that have been conducted with this model.

Nevertheless, information from GL261 and other subcutaneous tumour models will be applied to the orthotopic brain models in future experiments.

The effects of lipoglycan related compounds on inflammatory immune responses

Contributors: Dr William John Martin, Deborah Knight, Odette Shaw, Dr Ian Hermans, Dr Jacquie Harper, collaborating with Dr Phillip Rendle & Dr Gavin Painter (IRL), Assoc Prof Sarah Hook, Prof Thomas Rades & Dr David Larsen (University of Otago) and Prof Bryce Buddle (AgResearch)

In 2008 a multidisciplinary drug discovery initiative was launched in collaboration with Industrial Research Limited, University of Otago and AgResearch, to develop and test novel carbohydrate-lipid based compounds for their potential to either enhance or suppress inflammatory immune responses.

A focus of this programme is the identification of compounds with adjuvant properties that can be used to enhance tumour-specific immune responses for the development of more effective cell-mediated vaccines.

Lipoglycan related compounds are also being evaluated for their potential to suppress inappropriate inflammatory responses such as those seen in asthma and allergy.

The role of langerin+ CD8+ dendritic cells in priming immune responses to BCG

Contributors: Kelly Prendergast, Dr Ian Hermans, Dr Troels Petersen, Dr Joanna Kirman

Considerable effort is now being directed at vaccination strategies that target antigens to dendritic cells with known function. An interesting subset of dendritic cells expresses the cell surface markers langerin and CD8. This DC subset is found in the marginal zone of the spleen and is thought to play an important role in screening the blood for systemic antigens.

Dr Kirman’s Infectious Diseases Group is working with Dr Troels Petersen and colleagues to determine if these cells can prime T cell immune responses against intravenous BCG infection - a model of systemic bacterial infection.

They will also investigate whether langerin+ CD8+ DCs play a role in controlling bacterial BCG burden and reactivating memory T cells.

The outcome of this work will be a greater understanding of the dendritic cell types involved in mediating immunity against mycobacteria.

Publications

2012

Cheong S, Ferguson P, Hermans IF, Jameson GNL, Prabakar S, Herman DAJ, Tilley RD (2012) Synthesis and stability of highly crystalline and stable iron/iron oxide core/shell nanoparticles for biomedical applications. ChemPlusChem, 77:135-40

Dangerfield EM, Cheng JM, Knight DA, Weinkove R, Dunbar PR, Hermans IF, Timmer MS, Stocker BL (2012) Species-specific activity of glycolipid ligands for invariant NKT cells. Chembiochem, 13:1349-56

Dickgreber N, Farrand KJ, van Panhuys N, Knight DA, McKee SJ, Chong ML, Miranda-Hernandez S, Baxter AG, Locksley RM, Le Gros G, Hermans IF (2012) Immature murine NKT cells pass through a stage of developmentally programmed innate IL-4 secretion. J Leukoc Biol, [Epub ahead of print]

Hunn MK, Farrand KJ, Broadley KW, Weinkove R, Ferguson P, Miller RJ, Field CS, Petersen T, McConnell MJ, Hermans IF Vaccination with irradiated tumor cells pulsed wtih an adjuvant that stimulates NKT cells is an effective treatment for glioma. Clin Cancer Res, 2012 Nov 12 [Epub ahead of print]

McKee SJ, Young VL, Clow F, Hayman CM, Baird MA, Hermans IF, Young SL, Ward VK (2012) Virus-like particles and α-galactosylceramide form a self-adjuvanting composite particle that elicits anti-tumour immune responses. J Control Release, 159:338-45

Weinkove R, Brooks CR, Carter JM, Hermans IF, Ronchese F (2012) Functional invariant natural killer T cell and CD1d axis in chronic lymphocytic leukemia: implications for immunotherapy. Haematologica [Epub ahead of print]

2011

Ainge GD, Martin WJ, Compton BJ, Hayman CM, Larsen DS, Yoon S-i, Wilson IA, Harper JL, Painter GF (2011) Synthesis and toll-like receptor 4 (TLR4) activity of phosphatidylinositol dimannoside analogues. J Med Chem, 54:7268-79

Broadley KW, Hunn MK, Farrand KJ, Price KM, Grasso C, Miller RJ, Hermans IF, McConnell MJ (2011) Side population is not necessary or sufficient for a cancer stem cell phenotype in glioblastoma multiforme. Stem Cells, 29:452-61

Cheng JM, Chee SH, Knight DA, Acha-Orbea H, Hermans IF, Timmer MS, Stocker BL (2011) An improved synthesis of danyslated alpha-galactosylceramide and its use as a fluorescent probe for the monitoring of glycolipid uptake by cells. Carbohydr Res, 346:914-26

Cheong S, Ferguson P, Feindel KW, Hermans IF, Callaghan PT, Meyer C, Slocombe A, Su CH, Cheng FY, Yeh CS, Ingham B, Toney MF, Tilley RD (2011) Simple synthesis and functionalization of iron nanoparticles for magnetic resonance imaging. Angew Chem Int Ed Engl, 50:4206-9

Girvan RC, Knight DA, O'Loughlin CJ, Hayman CM, Hermans IF, Webster GA (2011) MIS416, a non-toxic microparticle adjuvant derived from Propionibacterium acnes comprising immunostimulatory muramyl dipeptide and bacterial DNA promotes cross-priming and Th1 immunity. Vaccine, 29:545-57

Herman DA, Ferguson P, Cheong S, Hermans IF, Ruck BJ, Allan KM, Prabakar S, Spencer JL, Lendrum CD, Tilley RD (2011) Hot-injection synthesis of iron/iron oxide core/shell nanoparticles for T(2) contrast enhancement in magnetic resonance imaging. Chem Commun (Camb), 47:9221-3

Petersen TR, Sika-Paotonu D, Knight DA, Simkins HM, Hermans IF (2011) Exploiting the role of endogenous lymphoid-resident dendritic cells in the priming of NKT cells and CD8+ T cells to dendritic cell-based vaccines. PLoS One, 6:e17657

Simkins HM, Hyde E, Farrand KJ, Ong ML, Degli-Esposti MA, Hermans IF, Ronchese F (2011) Administration of alpha-galactosylceramide impairs the survival of dendritic cell subpopulations in vivo. J Leukoc Biol, 89:753-62

2010

Brooks CR, Weinkove R, Hermans IF, van Dalen CJ, Douwes J (2010) Invariant natural killer T cells and asthma: immunologic reality or methodologic artifact? J Allergy Clin Immunol, 126:882-5

Petersen TR, Dickgreber N, Hermans IF (2010) Tumor antigen presentation by dendritic cells. Crit Rev Immunol, 30:345-86. Review.

Petersen TR, Sika-Paotonu D, Knight DA, Dickgreber N, Farrand KJ, Ronchese F, Hermans IF (2010) Potent anti-tumour responses to immunization with dendritic cells loaded with tumor tissue and an NKT cell ligand. Immunol Cell Biol, 88:596-604

2009

Dickgreber N, Stoitzner P, Bai Y, Price KM, Farrand KJ, Manning K, Angel CE, Dunbar PR, Ronchese F, Fraser JD, Bäckström BT, Hermans IF (2009) Targeting antigen to MHC class II molecules promotes cross-presentation and enhances immunotherapy. J Immunol, 182:1260-9

Farrand KJ, Dickgreber N, Stoitzner P, Ronchese F, Petersen TR, Hermans IF (2009) Langerin+CD8alpha+ dendritic cells are critical for cross-priming and IL-12 production in response to systemic antigens. J Immunol, 183:7732-42

Tripp CH, Sparber F, Hermans IF, Romani N, Stoitzner P (2009) Glycolipids injected into the skin are presented to NKT cells in the draining lymph node independently of migratory skin dendritic cells. J Immunol, 182:7644-54

2008

Andrew KA, Simkins HMA, Witzel S, Perret R, Hudson J, Hermans IF, Ritchie DS, Yang J, Ronchese F (2008) Dendritic cells treated with lipopolysaccharide up-regulate serine protease inhibitor 6 and remain sensitive to killing by cytotoxic T lymphocytes in vivo. J Immunol, 181:8356-62

Stoitzner P, Green LK, Jung JY, Price KM, Tripp CH, Malissen B, Kissenpfennig A, Hermans IF, Ronchese F (2008) Tumor immunotherapy by epicutaneous immunization requires Langerhans cells. J Immunol, 180:1991-8

2007

Hermans IF, Silk JD, Gileadi U, Masri SH, Shepherd D, Farrand KJ, Salio M, Cerundolo V (2007) Dendritic cell function can be modulated through the cooperative actions of TLR ligands and invariant NKT cells. J Immunol, 178:2721-9

Matthews KE, Qin JS, Yang J, Hermans IF, Palmowski MJ, Cerundolo V, Ronchese F (2007) Increasing the survival of dendritic cells in vivo does not replace the requirement for CD4+ T cell help during primary CD8+ T cell responses. J Immunol, 179:5738-47

2006

Lee A, Farrand KJ, Dickgreber N, Hayman CM, Jürs S, Hermans IF, Painter GF (2006) Novel synthesis of -galactosyl ceramides and confirmation of their powerful NKT cell agonist activity. Carbohydrate Research 341: 2785-2798

Matthews KE, Hermans IF, Roberts JM, Ching LM, Ronchese F (2006) 5,6-Dimethylxanthenone-4-acetic acid treatment of a non-immunogenic tumour does not synergize with active or passive CD8+ T-cell immunotherapy. Immunol Cell Biol, 84:383-9

2005

Yang J, Huck SP, McHugh RS, Hermans IF, Ronchese F (2005) Perforin-dependent elimination of dendritic cells regulates the expansion of antigen-specific CD8+ T cells in vivo. Proc Natl Acad Sci USA, 103:147-52

2004

Silk JD, Hermans IF, Gileadi U, Chong TW, Shepherd D, Salio M, Mathew B, Schmidt RR, Lunt SJ, Williams KJ, Stratford IJ, Harris AL, Cerundolo V (2004) Utilizing the adjuvant properties of CD1d-dependent NK T cells in T cell-mediated immunotherapy. J Clin Invest, 114:1800-11

Ritchie DS, Yang J, Hermans IF, Ronchese F (2004) B-Lymphocytes activated by CD40 ligand induce an antigen-specific anti-tumour immune response by direct and indirect activation of CD8(+) T-cells. Scand J Immunol, 60:543-51

Salio M, Palmowski MJ, Atzberger A, Hermans IF, Cerundolo V (2004) CpG-matured murine plasmacytoid dendritic cells are capable of in vivo priming of functional CD8 cell responses to endogenous but not exogenous antigens. J Exp Med, 199:567-79

Hermans IF, Silk JD, Yang J, Palmowski MJ, Gileadi U, McCarthy C, Salio M, Ronchese F, Cerundolo V (2004) The VITAL assay: a versatile fluorometric technique for assessing CTL- and NKT-mediated cytotoxicity against multiple targets in vitro and in vivo. J Immunol Methods, 285:25-40

Cerundolo V, Hermans IF, Salio M (2004) Dendritic cells: a journey from laboratory to clinic. Nat Immunol, 5:7-10