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Increased expression of HSP70 by colon cancer cells is not always associated with access to the dendritic cell cross-presentation pathway

Abstract

Dendritic cells (DCs) are highly specialized antigen-presenting cells endowed with the unique ability to not only present exogenous antigens upon exposure to MHC II, but also to cross-present these upon exposure to MHC I. This property was exploited to generate the tumor-specific CD8 cytotoxic lymphocyte (CTL) response in DCs-based cancer vaccine protocols. In this context, the source of tumor antigens remains a critical challenge. A crude tumor in the context of danger signals is believed to represent an efficient source of tumor antigens (TAs) for DCs loading. In our previous work, increased DCs cross-presentation of antigens from necrotic gastric carcinoma cells paralleled up-regulation of the heat shock protein hsp70. We studied the expression of hsp70 on primary colon carcinoma cells and its relevance in the cross-priming of anti-tumor CTL by tumor-loaded DCs. Hsp70 was expressed on all three of the tumors studied, but was never detected in the peritumoral normal mucosa (NM). The uptake of the tumor induced a trend towards down-modulation of the monocyte-specific marker CD14, but had no effect on the chemokine receptors CCR4 and CCR7. The IFN-γ enzyme-linked immunospot assay (ELIspot) showed cross-priming of CTL by tumor-loaded but not NM-loaded DCs in four of the six cases studied. The CTL response generated in DC+tumor cultures was directed towards the tumor, but not towards NM, and it was characterized by refractoriness to polyclonal (Ca ionophores, PKC activators) stimuli. Of the three CTL-generating tumors, only one expressed hsp70. This data indicates a tumor-specific expression of hsp70, but does not support its relevance in the DC cross-presentation of TAs.

Abbreviations

Ag:

antigen

AJCC:

American Joint Committee on Cancer

APC:

antigen-presenting cell

CCR:

chemokines receptor

CD4:

cluster of differentiation 4

CD8:

cluster of 8

CTL:

cytotoxic T lymphocyte

DCs:

dendritic cell

DMSO:

dimethylsulfoxide

ELIspot:

enzyme linked immunospot

HSP differentiation:

heat shock protein

IFN:

interferon

Il:

interleukin

MHC:

major histocompatibility complex

NM:

normal mucosa

PBMC:

peripheral blood mononuclear cell

RIPA buffer:

radioimmunoprecipitation buffer

TAs:

tumor associated antigen

TNF:

tumor necrosis factor

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Correspondence to Lina Matera.

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Matera, L., Forno, S., Galetto, A. et al. Increased expression of HSP70 by colon cancer cells is not always associated with access to the dendritic cell cross-presentation pathway. Cell Mol Biol Lett 12, 268–279 (2007). https://doi.org/10.2478/s11658-007-0001-6

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  • DOI: https://doi.org/10.2478/s11658-007-0001-6

Key words

  • Dendritic cells
  • Cytotoxic T lymphocyte
  • Colon cancer
  • Necrosis
  • Heat shock proteins