Skip to main content
Download PDF

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

Cellular & Molecular Biology Letters volume 12, pages 268–279 (2007)Cite this article

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

References

  1. Jemal, A., Murray, T., Samuels, A., Ghafoor, A., Ward, E. and Thun, M.J. Cancer statistics. CA Cancer J. Clin. 49 (1999) 202–219.

    Google Scholar 

  2. Midgley, R. and Kerr, D. Immunotherapy for colorectal cancer: a challenge to clinical trial design. Lancet Oncol. 1 (2000) 159–168.

    Article  CAS  PubMed  Google Scholar 

  3. Coutinho, A.K. and Rocha Lima, C.M. Metastatic colorectal cancer: systemic treatment in the new millennium. Cancer Control 10 (2003) 224–238.

    PubMed  Google Scholar 

  4. Mocellin, S., Rossi, C.R., Lise, M. and Nitti, D. Colorectal cancer vaccines: principles, results, and perspectives. Gastroenterology 127 (2004) 1821–1837.

    Article  CAS  PubMed  Google Scholar 

  5. d’Entremont, T.S. and Sun, W. Recent advances in colorectal cancer therapy. Cancer Biol. Ther. 2 (2003) 6–13.

    PubMed  Google Scholar 

  6. Van Der Bruggen, P., Zhang, Y., Chaux, P., Stroobant, V., Panichelli, C., Schultz, E.S., Chapiro, J., Van Den Eynde, B.J., Brasseur, F. and Boon, T. Tumor-specific shared antigenic peptides recognized by human T cells. Immunol. Rev. 188 (2002) 51–64.

    Article  Google Scholar 

  7. Figdor, C.G., de Vries, I.J., Lesterhuis, W.J. and Melief, C.J.M. Dendritic cell immunotherapy: mapping the way. Nat. Med. 10 (2004) 475–480.

    Article  CAS  PubMed  Google Scholar 

  8. Banchereau, J. and Steinman, R. Dendritic cells and the control of immunity. Nature 392 (1998) 245–252.

    Article  CAS  PubMed  Google Scholar 

  9. Ridge, J.P., Di Rosa, F. and Matzinger, P. A conditioned dendritic cell can be a temporal bridge between a CD4+ T-helper and T-killer cell. Nature 393 (1998) 474–478.

    Article  CAS  PubMed  Google Scholar 

  10. Parmiani, G., Castelli, C., Dalerba, P., Mortarini, R., Rivoltini, L., Marincola, F.M. and Anichini, A. Cancer immunotherapy with peptide-based vaccines: what have we achieved? Where are we going? J. Natl. Cancer Inst. 94 (2002) 805–809.

    CAS  PubMed  Google Scholar 

  11. Schnurr, M., Scholz, C., Rothenfusser, S., Galambos, P., Dauer, M., Robe, J., Endres, S. and Eigler, A. Apoptotic pancreatic tumor cells are superior to cell lysates in promoting crosspriming of cytotoxic T cells and activate NK and gammadelta T cells. Cancer Res. 62 (2002) 2347–2352.

    CAS  PubMed  Google Scholar 

  12. Kotera, Y., Shimizu K. and Mulé, J.J. Comparative analysis of necrotic and apoptotic tumor cells as a source of antigens in dendritic cell-based immunization. Cancer Res. 61 (2001) 8105–8109.

    CAS  PubMed  Google Scholar 

  13. Pietra, G., Mortarini, R., Parmiani, G. and Anichini, A. Phases of apoptosis of melanoma cells, but not of normal melanocytes, differently affect maturation of myeloid dendritic cells. Cancer Res. 61 (2001) 8218–8226.

    CAS  PubMed  Google Scholar 

  14. Schnurr, M., Galambos, P., Scholz, C., Then, F., Dauer, M., Endres, S. and Eigler, A. Tumor cell lysate-pulsed human dendritic cells induce a T-cell response against pancreatic carcinoma cells: an in vitro model for the assessment of tumor vaccines. Cancer Res. 61 (2001) 6445–6450.

    CAS  PubMed  Google Scholar 

  15. Hoffmann, T.K., Meidenbauer. N., Dworacki, G., Kanaya, H. and Whiteside, T.L. Generation of tumor-specific T lymphocytes by cross-priming with human dendritic cells ingesting apoptotic tumor cells. Cancer Res. 60 (2000) 3542–3549.

    CAS  PubMed  Google Scholar 

  16. Albert, M.L., Sauter, B. and Bhardwaj, N. Dendritic cells acquire antigens from apoptotic cells and induce class I-restricted CTLs. Nature 392 (1998) 86–89.

    Article  CAS  PubMed  Google Scholar 

  17. Gallucci, S., Lolkema, M. and Matzinger, P. Natural adjuvants: endogenous activators of dendritic cells. Nat. Med. 11 (1999) 1249–1255.

    Article  Google Scholar 

  18. Basu, S., Binder, R., Suto, R., Anderson, K. and Srivastava, P. Necrotic but not apoptotic cell death releases heat shock protein expression, which deliver a partial maturation signal to dendritic cells and activate the NK-kB pathway. Int. Immunol. 12 (2000) 1539–1546.

    Article  CAS  PubMed  Google Scholar 

  19. Melcher, A., Todryk, S., Hardwick, N., Ford, M., Jacobson, M. and Vile, R.G. Tumor immunogenicity is determined by the mechanism of cell death via induction of heat shock protein expression. Nat. Med. 4 (1998) 581–587.

    Article  CAS  PubMed  Google Scholar 

  20. Hongzhen S., Tinghua C., Connolly, J.E., Monnet, L., Bennett, L., Chapel, S., Bagnis, C., Mannoni, P., Davoust, J., Palucka, A.K. and Banchereau, J. Hyperthermia Enhances CTL Cross-Priming. J. Immunol. 176 (2006) 2134–2141.

    Google Scholar 

  21. Milani, V., Noessner, E., Ghose, S., Kuppner, M., Ahrens, B., Scharner, A., Gastpar, R. and Issels, R.D. Heat shock protein 70: role in antigen presentation and immune stimulation. Int. J. Hyperthermia 18 (2002) 563–575.

    Article  CAS  PubMed  Google Scholar 

  22. Kuppner, M., Gastpar, R., Gelwer, S., Noessner, E., Ochmann, O., Scharner, A. and Issels, R.D. The role of heath shock protein (hsp70) in dendritic cell maturation: hsp70 induces the maturation of immature dendritic cells but reduces DC differentiation from monocytes precursors. J. Eur. Immunol. 31 (2001) 1602–1609.

    Article  CAS  Google Scholar 

  23. Galetto, A., Buttiglieri, S., Forno, S., Moro, F., Mussa, A. and Matera, L. Drug-and cell-mediated antitumor cytotoxicities modulate crosspresentation of tumor antigens by myeloid dendritic cells. Anti-Cancer Drugs 14 (2003) 883–843.

    Google Scholar 

  24. Galetto, A., Contarini, M., Sapino, A., Cassoni, P., Consalvo, E., Forno, S., Pezzi, C., Barnaba, V., Mussa, A. and Matera, L. Ex-vivo host response to gastrointestinal cancer cells presented by autologous dendritic cells. J. Surgical. Res. 100 (2001) 32–38.

    Article  CAS  Google Scholar 

  25. Buttiglieri, S., Galetto, A., Forno, S., De Andrea, M. and Matera, L. Influence of drug-induced apoptotic death on processing and presentation of tumor antigens by dendritic cells. Int. J. Cancer 106 (2003) 516–520.

    Article  CAS  PubMed  Google Scholar 

  26. Luft, T., Jefford, M., Luetjens, P., Toy, T., Hochrein, H., Masterman, K.A., Maliszewski, C., Shortman, K., Cebon, J. and Maraskovsky, E. Functionally distinct dendritic cell (DC) populations induced by physiologic stimuli: prostaglandin E(2) regulates the migratory capacity of specific DC subsets. Blood 100 (2002) 1362–1372.

    Article  CAS  PubMed  Google Scholar 

  27. Chang, C.C. and Ferrone, S. Immune selective pressure and HLA class I antigen defects in malignant lesions. Cancer. Immunol. Immunother. (2006) Epub ahead of print.

  28. Vakkila, J. and Lotze, M.T. Inflammation and necrosis promote tumour growth. Nat. Rev. Immunol. 8 (2004) 641–648.

    Article  Google Scholar 

  29. Rovere-Querini, P., Capobianco, A., Scaffidi, P., Valentinis, B., Catalanotti, F., Giazzon, M., Dumitriu, I.E., Muller, S., Iannacone, M., Traversari, C., Bianchi, M.E. and Manfredi, A.A. HMGB1 is an endogenous immune adjuvant released by necrotic cells. EMBO Rep. 8 (2004) 825–830.

    Article  Google Scholar 

Download references

Author information

Author notes

  1. Alessandra Galetto

    Present address: University of East Piedmont “Amedeo Avogadro”, Novara, Italy

Authors and Affiliations

  1. Deptartment of Internal Medicine, University of Turin, Italy

    Lina Matera, Sarah Forno, Alessandra Galetto & Stefano Garetto

  2. Unit of Liver Transplantation, Molinette Hospital, Turin, Italy

    Francesco Moro

  3. S.C.D.U. of Surgical Oncology, University of Turin, Italy

    Antonio Mussa

Authors
  1. Lina Matera
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Sarah Forno
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Alessandra Galetto
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Francesco Moro
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Stefano Garetto
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Antonio Mussa
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Lina Matera.

Rights and permissions

Reprints and Permissions

About this article

Cite this article

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

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

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

Key words

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

Cellular & Molecular Biology Letters

ISSN: 1689-1392

Contact us