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Interaction of human heat shock protein 70 with tumor-associated peptides

  • Maya J. Pandya , Henriette Bendz , Florian Manzenrieder , Elfriede Noessner , Horst Kessler , Johannes Buchner and Rolf D. Issels
Published/Copyright: April 1, 2009
Biological Chemistry
From the journal Volume 390 Issue 4

Abstract

Molecular chaperones of the heat shock protein 70 (Hsp70) family play a crucial role in the presentation of exogenous antigenic peptides by antigen-presenting cells (APCs). In a combined biochemical and immunological approach, we characterize the biochemical interaction of tumor-associated peptides with human Hsp70 and show that the strength of this interaction determines the efficacy of immunological cross-presentation of the antigenic sequences by APCs. A fluorescein-labeled cytosolic mammalian Hsc70 binding peptide is shown to interact with human Hsp70 molecules with high affinity (Kd=0.58 μm at 25°C). Competition experiments demonstrate weaker binding by Hsp70 of antigenic peptides derived from the tumor-associated proteins tyrosinase (Kd=32 μm) and melanoma antigen recognized by T cells (MART-1) (Kd=2.4 μm). Adding a peptide sequence (pep70) with high Hsp70 binding affinity (Kd=0.04 μm) to the tumor-associated peptides enables them to strongly interact with Hsp70. Presentation of tumor-associated peptides by B cells resulting in T cell activation in vitro is enhanced by Hsp70 when the tumor-associated peptides contain the Hsp70 binding sequence. This observation has relevance for vaccine design, as augmented transfer of tumor-associated antigens to APCs is closely linked to the vaccine's efficacy of T cell stimulation.


Corresponding author

We thank Stefanie Hufnagl for assistance with protein purification, Helmut Krause for performing mass spectrometry, Daniel Neumaier and Anna Brandl for the T and B cell cultures. The clone for human Hsp70 was kindly provided by Matthias Mayer, Molecular Biology Center, University of Heidelberg, Germany. We are grateful to Stefan Walter for helpful discussions. This work was supported by the Deutsche Forschungsgemeinschaft SFB455 (to E.N. and R.D.I.).

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Received: 2008-10-28
Accepted: 2009-1-19
Published Online: 2009-04-01
Published in Print: 2009-04-01

©2009 by Walter de Gruyter GmbH & Co. KG

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