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Cellular & Molecular Biology Letters logoLink to Cellular & Molecular Biology Letters
. 2007 Jan 19;12(2):268–279. doi: 10.2478/s11658-007-0001-6

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

Lina Matera 1,✉, Sarah Forno 1, Alessandra Galetto 1, Francesco Moro 2, Stefano Garetto 1, Antonio Mussa 3
PMCID: PMC6275593  PMID: 17235439

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.

Key words: Dendritic cells, Cytotoxic T lymphocyte, Colon cancer, Necrosis, Heat shock proteins

Full Text

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Abbreviations used

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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