A novel method to identify and characterise peptide mimotopes of heat shock protein 70-associated antigens.
The heat shock protein, Hsp70, has been shown to play an important role in tumour immunity. Vaccination with Hsp70-peptide complexes (Hsp70-PCs), isolated from autologous tumour cells, can induce protective immune responses. We have developed a novel method to identify synthetic mimic peptides of Hsp70-PCs and to test their ability to activate T-cells. Peptides (referred to as "recognisers") that bind to Hsp70-PCs from the human breast carcinoma cell line, MDA-MB-231, were identified by bio-panning a random peptide M13 phage display library. Synthetic recogniser peptides were subsequently used as bait in a reverse bio-panning experiment to identify potential Hsp70-PC mimic peptides. The ability of the recogniser and mimic peptides to prime human lymphocyte responses against tumour cell antigens was tested by stimulating lymphocytes with autologous peptide-loaded monocyte-derived dendritic cells (DCs). Priming and subsequent stimulation with either the recogniser or mimic peptide resulted in interferon-gamma (IFN-gamma) secretion by the lymphocytes. Furthermore, DCs loaded with Hsp70, Hsp70-PC or the recogniser or the mimic peptide primed the lymphocytes to respond to soluble extracts from breast cells. These results highlight the potential application of synthetic peptide-mimics of Hsp70-PCs, as modulators of the immune response against tumours.
Arnaiz
Blanca
B
Moyne Institute for Preventive Medicine, Department of Microbiology, University of Dublin, Trinity College, Dublin 2, Ireland.
Madrigal-Estebas
Laura
L
Institute of Immunology & Department of Biology, National University of Ireland, Maynooth, Co. Kildare, Ireland.
Todryk
Stephen
S
Centre for Clinical Vaccinology and Tropical Medicine, Churchill Hospital, Oxford OX3 7LJ, UK.
James
Tharappel C
TC
Moyne Institute for Preventive Medicine, Department of Microbiology, University of Dublin, Trinity College, Dublin 2, Ireland.
Doherty
Derek G
DG
Institute of Immunology & Department of Biology, National University of Ireland, Maynooth, Co. Kildare, Ireland.
Bond
Ursula
U
Moyne Institute for Preventive Medicine, Department of Microbiology, University of Dublin, Trinity College, Dublin 2, Ireland.