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A novel method to identify and characterise peptide mimotopes of heat shock protein 70-associated antigens

Blanca Arnaiz1, Laura Madrigal-Estebas2, Stephen Todryk3, Tharappel C James1, Derek G Doherty2 and Ursula Bond1*

  • * Corresponding author: Ursula Bond

  • † Equal contributors

Author Affiliations

1 Moyne Institute for Preventive Medicine, Department of Microbiology, University of Dublin, Trinity College, Dublin 2, Ireland

2 Institute of Immunology & Department of Biology, National University of Ireland, Maynooth, Co. Kildare, Ireland

3 Centre for Clinical Vaccinology and Tropical Medicine, Churchill Hospital, Oxford OX3 7LJ, UK

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Journal of Immune Based Therapies and Vaccines 2006, 4:2  doi:10.1186/1476-8518-4-2

Published: 8 April 2006


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-γ (IFN-γ) 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.