Differential capacity of chaperone-rich lysates in cross-presenting human endogenous and exogenous melanoma differentiation antigens

Elke Bleifuss, Henriette Bendz, Birgit Sirch, Sylvia Thompson, Anna Brandl, Valeria Milani, Michael W. Graner, Ingo Drexler, Maria Kuppner, Emmanuel Katsanis, Elfriede Noessner, Rolf Dieter Issels

Research output: Contribution to journalArticlepeer-review

9 Scopus citations

Abstract

The goal of immune-based tumor therapies is the activation of immune cells reactive against a broad spectrum of tumor-expressed antigens. Vaccines based on chaperone proteins appear promising as these proteins naturally exist as complexes with various protein fragments including those derived from tumor-associated antigens. Multi-chaperone systems are expected to have highest polyvalency as different chaperones can carry distinct sets of antigenic fragments. A free-solution isoelectric focusing (FS-IEF) technique was established to generate chaperone-rich cell lysates (CRCL). Results from murine systems support the contention that CRCL induce superior anti-tumor responses than single chaperone vaccines. We established an in vitro model for human melanoma to evaluate the capacity of CRCL to transfer endogenously expressed tumor antigens to the cross-presentation pathway of dendritic cells (DC) for antigen-specific T cell stimulation. CRCL prepared from human melanoma lines contained the four major chaperone proteins Hsp/Hsc70, Hsp90, Grp94/gp96 and calreticulin. The chaperones within the melanoma cell-derived CRCL were functionally active in that they enhanced cross-presentation of exogenous peptides mixed into the CRCL preparation. Superior activity was observed for Hsp70-rich CRCL obtained from heat-stressed melanoma cells. Despite the presence of active chaperones, melanoma cell-derived CRCL failed to transfer endogenously expressed melanoma-associated antigens to DC for cross-presentation and cytotoxic T cell (CTL) recognition, even after increasing intracellular protein levels of tumor antigen or chaperones. These findings reveal limitations of the CRCL approach regarding cross-presentation of endogenously expressed melanoma-associated antigens. Yet, CRCL may be utilized as vehicles to enhance the delivery of exogenous antigens for DC-mediated cross-presentation and T cell stimulation.

Original languageEnglish (US)
Pages (from-to)623-637
Number of pages15
JournalInternational Journal of Hyperthermia
Volume24
Issue number8
DOIs
StatePublished - Dec 22 2008

Keywords

  • Chaperone-rich cell lysate (CRCL)
  • Cross-presentation
  • Dendritic cells
  • Heat shock proteins
  • Heat shock treatment
  • Melanoma-associated antigens

ASJC Scopus subject areas

  • Physiology
  • Physiology (medical)
  • Cancer Research

Fingerprint Dive into the research topics of 'Differential capacity of chaperone-rich lysates in cross-presenting human endogenous and exogenous melanoma differentiation antigens'. Together they form a unique fingerprint.

Cite this