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Identification of potential HLA class I and class II epitope precursors associated with heat shock protein 70 (HSPA)

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Cell Stress and Chaperones Aims and scope

Abstract

Heat shock protein 70 (HSPA) is a molecular chaperone which has been suggested to shuttle human leukocyte antigen (HLA) epitope precursors from the proteasome to the transporter associated with antigen processing. Despite the reported observations that peptides chaperoned by HSPA are an effective source of antigens for cross-priming, little is known about the peptides involved in the process. In this study, we investigated the possible involvement of HSPA in HLA class I or class II antigen presentation and analysed the antigenic potential of the associated peptides. HSPA was purified from CCRF-CEM and K562 cell lines, and using mass spectrometry techniques, we identified 44 different peptides which were co-purified with HSPA. The affinity of the identified peptides to two HSPA isoforms, HSPA1A and HSPA8, was confirmed using a peptide array. Four of the HSPA-associated peptides were matched with 13 previously reported HLA epitopes. Of these 13 peptides, nine were HLA class I and four were HLA class II epitopes. These results demonstrate the association of HSPA with HLA class I and class II epitopes, therefore providing further evidence for the involvement of HSPA in the antigen presentation process.

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Acknowledgements

We would like to thank to Dr. Joe Gray from the Pinnacle Lab, the Institute of Cell and Molecular Biosciences, Newcastle University, Newcastle upon Tyne, UK for the service and extensive help with the mass spectrometric analysis. This work was supported by grant from the Leukaemia Research Fund, UK [04070]; “Trans-Net” grant from the European Commission [MRTN-CT-2004-512253]; German Research Foundation (DFG) [MU1238/7-2] and the German Federal Ministry of Education and Research (BMBF) MOBITUM [01EZ0826] grants.

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Authors and Affiliations

  1. Haematological Sciences, Institute of Cellular Medicine, Medical School, Framlington Place, Newcastle University, Newcastle upon Tyne, NE2 4HH, UK

    Pawel Stocki, Xiao N. Wang & Anne M. Dickinson

  2. School of Biomedical Sciences, Newcastle University, Newcastle upon Tyne, NE2 4AB, UK

    Nicholas J. Morris

  3. The Beatson Institute for Cancer Research, Cancer Research UK, Glasgow, G61 1BD, UK

    Christian Preisinger & Walter Kolch

  4. Department of Radiotherapy/Radiooncology, Technische Universität München, 81675, Munich, Germany

    Gabriele Multhoff

  5. Institute of Pathology, Helmholtz Zentrum München, 85764, Neuherberg, Germany

    Gabriele Multhoff

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  1. Pawel Stocki
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Correspondence to Anne M. Dickinson.

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Stocki, P., Morris, N.J., Preisinger, C. et al. Identification of potential HLA class I and class II epitope precursors associated with heat shock protein 70 (HSPA). Cell Stress and Chaperones 15, 729–741 (2010). https://doi.org/10.1007/s12192-010-0184-z

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  • DOI: https://doi.org/10.1007/s12192-010-0184-z

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