- To identify new HLA-classes I and II restricted antigens (Ags) from human tumours of different histology and to define the role of CD4+ T cells in induction and maintenance of effective cytotoxic T lymphocyte (CTL) responses.
- To evaluate the role of chaperon proteins, including HSP, in inducing tumour specific T cell-mediated immune responses in human and murine cancers both in vitro and in vivo, comparing this approach with a different antigen-delivery system (recombinant filamentous phages).
- To molecularly define processing and presentation pathways of chaperon proteins, including cloning of receptors involved in their specific uptake by antigen presenting cells (APC).
- To engineer tumour cell and/or APC to overexpress chaperon proteins or their receptors as tools for effective immunisation strategies.
- To design pilot phase I-II studies for vaccine development in cancer patients.
DESCRIPTION OF THE WORK
- We will identify new tumour-derived HLA-class I and II-binding peptides by several approaches (cDNA or expression libraries, differential screening by micro-array technologies, etc). These peptides will be complexed with chaperon proteins for in vitro and in vivo immunological studies. New Ags will be also identified from tumour-derived chaperon proteins either by direct biochemical approaches (peptide elution and mass-spectrometry sequencing) or by cDNA cloning using for the screening procedures chaperon-induced tumour-specific T cells.
- The cellular interaction among dendritic cells (DC), CD4+ T cells and CTL will be studied in different in vitro systems. Attempts will be made to identify the signal involved in the induction of anti-tumour CTL in human system.
- The role of chaperon proteins in Ag processing and presentation will be analysed in tumour systems (in vitro and in vivo), and will include cloning of the receptors expressed by APC and mediating specific internalisation of chaperon/peptides complexes. The ability of native tumour-derived chaperon proteins to induce and modulate T cell specificities and functions will be studied in different human cancers, including melanoma, colon, renal and ovarian carcinomas.
- The assessment of the ability of antigenic peptide and chaperon proteins to function as natural adjuvant in potentiating tumour-specific immune responses, will be carried out as follows:
- 4a. Engineering tumour cells to overexpress HSP in native form or covalently linked to immunogenic peptides. Evaluate their immunogenicity in vitro (human and murine systems) and in vivo (HLA transgenic mice).
- 4b. Engineering APC to overexpress chaperon receptors in order to enhance their ability to process and present specific Ag.
- On the basis of the acquired knowledge, vaccines will be developed in pre-clinical models; phase I-II pilot clinical studies in cancer patients will be designed.
- Availability of new human unique and shared tumour Ags presented
by classes I and II HLA. Identification of immunogenic peptides
bound to HSP and mediating an immune response in cancer patients.
- Identification of the role of CD4+ T cells and definition
of essential factors to induce an effective anti-tumour CTL
- Identification of the pathway of Ag presentation by chaperon
proteins: and of their receptor expressed by APC.
- Engineered APC and optimisation of the function of chaperon
proteins in inducing an efficient T cell response: their use
as vaccines in a preclinical model.
- Design of phase I-II vaccination protocols for cancer patients.
Dr. Giorgio Parmiani
Unit of Immunotherapy of Human Tumour
Instituto Nazionale Tumori
Via Venezian 1
20133 Milano, Italy
Tel: +39 022390328
Fax: +39 022390630
Dr. Piergiuseppe De Berardinis
Institute of Protein Biochemistry and Enzymology
Via Marconi 10
80125 Napoli, Italy
Tel: +39 0817257228
Fax: +39 0812396525
Dr. Prof. Pierre G. Coulie
Cellular Genetics Unit
Université Catholique de Louvain
Avenue Hippocrate 74
1200 Brussels, Belgium
Tel: +32 27647599
Fax: +32 27647590
Prof. Frederic Triebel
Unité d'Immunologie Cellulaire
Institut Gustave Roussy
39, rue Camille Desmoulins
94805 Villejuif, France
Tel: +33 142114068
Fax: +33 142115268
Prof. Hans-Georg Rammensee
Interfakultäres Institut für Zellbiologie
Auf der Morgenstelle 15
72076 Tübingen, Germany
Tel: +49 70712980991
Fax: +49 7071295653
Prof. Cornelis JM Melief
LUMCC, Dpt. IHB-Bld 1, E3Q
Leiden University Medical Center
PO Box 2300 RC
9600 Leiden, The Netherlands
Tel: +31 715263800
Fax: +31 715216751
Prof. Rolf Kiessling
Dpt Oncology MTC, Karolinska Institute
CCK R8:01 Karolinska Hospital
17176 Stockholm, Sweden
Tel: +46 8321897
Fax: +46 8309195
Dr. Catia Traversari
Via Olgettina 58
20132 Milano, Italy
Tel: +39 0226434706
Fax: +39 0226434668