Cancer Immunology, Immunotherapy

, Volume 57, Issue 11, pp 1665–1673 | Cite as

Inefficient presentation of tumor-derived antigen by tumor-infiltrating dendritic cells

  • Patrizia StoitznerEmail author
  • Laura K. Green
  • Jae Y. Jung
  • Kylie M. Price
  • Haley Atarea
  • Bronwyn Kivell
  • Franca Ronchese
Original Article



Transplantable B16 melanoma is widely used as a tumor model to investigate tumor immunity. We wished to characterize the leukocyte populations infiltrating B16 melanoma tumors, and the functional properties of tumor-infiltrating dendritic cells (TIDC).

Materials and methods

We used the B16 melanoma cell line expressing ovalbumin protein (OVA) to investigate the phenotype and T cell stimulatory capacity of TIDC.


The majority of leukocytes in B16 melanoma were macrophages, which colocalized with TIDCs, B and T cells to the peripheral area of the tumor. Both myeloid and plasmacytoid DC populations were present within tumors. Most of these DCs appeared immature, but about a third expressed a mature phenotype. TIDCs did not present tumor-derived antigen, as they were unable to induce the proliferation of tumor-specific CD4+ and CD8+ T cells in vitro unless in the presence of specific peptides. Some presentation of tumor-derived antigen could be demonstrated in the tumor-draining lymph node using in vivo proliferation assays. However, while proliferation of CD8+ T cells was reproducibly demonstrated, no proliferation of CD4+ T cells was observed.


In summary, our data suggest that DCs in tumors have limited antigen-presenting function. Inefficient antigen presentation extends to the tumor-draining lymph node, and may affect the generation of antitumor immune responses.


Melanoma Dendritic cells Tumor immunity 



Dendritic cells


Tumor-infiltrating DC


Langerhans cells




T cell receptor


Monoclonal antibody




fluorescein isothiocyanate




Phosphate-buffered saline




Counts per minute


Carboxy-fluorescein diacetate succinimidyl ester




Peridinin chlorophyll A protein


Granulocyte/macrophages colony-stimulating factor





We thank Drs E. Lord, J.G. Frelinger and F. Carbone for generously providing cell lines and mouse strains used in this study, the staff of the Malaghan Experimental Research Facility for animal husbandry and care, and the staff of the Malaghan Institute for useful suggestions and discussion. This work was supported by research grants from the Health Research Council and Cancer Society of NZ, the Wellington Medical Research Foundation, and the Genesis Oncology Trust. PS was supported by the Erwin Schroedinger Auslandsstipendium from the Austrian Science Fund (FWF-J2479).


  1. 1.
    Barnden MJ, Allison J, Heath WR, Carbone FR (1998) Defective TCR expression in transgenic mice constructed using cDNA-based alpha- and beta-chain genes under the control of heterologous regulatory elements. Immunol Cell Biol 76:34–40PubMedCrossRefGoogle Scholar
  2. 2.
    Bell D, Chomarat P, Broyles D, Netto G, Harb GM, Lebecque S, Valladeau J, Davoust J, Palucka KA, Banchereau J (1999) In breast carcinoma tissue, immature dendritic cells reside within the tumor, whereas mature dendritic cells are located in peritumoral areas. J Exp Med 190:1417–1426PubMedCrossRefGoogle Scholar
  3. 3.
    Bennaceur K, Popa I, Portoukalian J, Berthier-Vergnes O, Peguet-Navarro J (2006) Melanoma-derived gangliosides impair migratory and antigen-presenting function of human epidermal Langerhans cells and induce their apoptosis. Int Immunol 18:879–886PubMedCrossRefGoogle Scholar
  4. 4.
    Berthier-Vergnes O, Gaucherand M, Peguet-Navarro J, Plouet J, Pageaux JF, Schmitt D, Staquet MJ (2001) Human melanoma cells inhibit the earliest differentiation steps of human Langerhans cell precursors but failed to affect the functional maturation of epidermal Langerhans cells. Br J Cancer 85:1944–1951PubMedCrossRefGoogle Scholar
  5. 5.
    Chiodoni C, Paglia P, Stoppacciaro A, Rodolfo M, Parenza M, Colombo MP (1999) Dendritic cells infiltrating tumors cotransduced with granulocyte/macrophage colony-stimulating factor (GM-CSF) and CD40 ligand genes take up and present endogenous tumor-associated antigens, and prime naive mice for a cytotoxic T lymphocyte response. J Exp Med 190:125–133PubMedCrossRefGoogle Scholar
  6. 6.
    Curiel TJ, Coukos G, Zou L, Alvarez X, Cheng P, Mottram P, Evdemon-Hogan M, Conejo-Garcia JR, Zhang L, Burow M, Zhu Y, Wei S, Kryczek I, Daniel B, Gordon A, Myers L, Lackner A, Disis ML, Knutson KL, Chen L, Zou W (2004) Specific recruitment of regulatory T cells in ovarian carcinoma fosters immune privilege and predicts reduced survival. Nat Med 10:942–949PubMedCrossRefGoogle Scholar
  7. 7.
    Hogquist KA, Jameson SC, Heath WR, Howard JL, Bevan MJ, Carbone FR (1994) T cell receptor antagonist peptides induce positive selection. Cell 76:17–27PubMedCrossRefGoogle Scholar
  8. 8.
    Huang AY, Golumbek P, Ahmadzadeh M, Jaffee E, Pardoll D, Levitsky H (1994) Role of bone marrow-derived cells in presenting MHC class I-restricted tumor antigens. Science 264:961–965PubMedCrossRefGoogle Scholar
  9. 9.
    Ishida T, Oyama T, Carbone DP, Gabrilovich DI (1998) Defective function of Langerhans cells in tumor-bearing animals is the result of defective maturation from hemopoietic progenitors. J Immunol 161:4842–51PubMedGoogle Scholar
  10. 10.
    Kusmartsev S, Gabrilovich DI (2002) Immature myeloid cells and cancer-associated immune suppression. Cancer Immunol Immunother 51:293–298PubMedCrossRefGoogle Scholar
  11. 11.
    Li M, Davey GM, Sutherland RM, Kurts C, Lew AM, Hirst C, Carbone FR, Heath WR (2001) Cell-associated ovalbumin is cross-presented much more efficiently than soluble ovalbumin in vivo. J Immunol 166:6099–6103PubMedGoogle Scholar
  12. 12.
    Lugade AA, Moran JP, Gerber SA, Rose RC, Frelinger JG, Lord EM (2005) Local radiation therapy of B16 melanoma tumors increases the generation of tumor antigen-specific effector cells that traffic to the tumor. J Immunol 174:7516–7523PubMedGoogle Scholar
  13. 13.
    Lyons AB (2000) Analysing cell division in vivo and in vitro using flow cytometric measurement of CFSE dye dilution. J Immunol Methods 243:147–154PubMedCrossRefGoogle Scholar
  14. 14.
    Marzo AL, Kinnear BF, Lake RA, Frelinger JJ, Collins EJ, Robinson BW, Scott B (2000) Tumor-specific CD4+ T cells have a major “post-licensing” role in CTL mediated anti-tumor immunity. J Immunol 165:6047–6055PubMedGoogle Scholar
  15. 15.
    Marzo AL, Lake RA, Lo D, Sherman L, McWilliam A, Nelson D, Robinson BW, Scott B (1999) Tumor antigens are constitutively presented in the draining lymph nodes. J Immunol 162:5838–5845PubMedGoogle Scholar
  16. 16.
    McLellan AD, Kapp M, Eggert A, Linden C, Bommhardt U, Brocker EB, Kammerer U, Kampgen E (2002) Anatomic location and T-cell stimulatory functions of mouse dendritic cell subsets defined by CD4 and CD8 expression. Blood 99:2084–2093PubMedCrossRefGoogle Scholar
  17. 17.
    Movassagh M, Spatz A, Davoust J, Lebecque S, Romero P, Pittet M, Rimoldi D, Lienard D, Gugerli O, Ferradini L, Robert C, Avril MF, Zitvogel L, Angevin E (2004) Selective accumulation of mature DC-Lamp+ dendritic cells in tumor sites is associated with efficient T-cell-mediated antitumor response and control of metastatic dissemination in melanoma. Cancer Res 64:2192–2198PubMedCrossRefGoogle Scholar
  18. 18.
    Nagaraj S, Gupta K, Pisarev V, Kinarsky L, Sherman S, Kang L, Herber DL, Schneck J, Gabrilovich DI (2007) Altered recognition of antigen is a mechanism of CD8+ T cell tolerance in cancer. Nat Med 13:828–835PubMedCrossRefGoogle Scholar
  19. 19.
    Ossendorp F, Toes RE, Offringa R, van der Burg SH, Melief CJ (2000) Importance of CD4(+) T helper cell responses in tumor immunity. Immunol Lett 74:75–79PubMedCrossRefGoogle Scholar
  20. 20.
    Pinzon-Charry A, Maxwell T, Lopez JA (2005) Dendritic cell dysfunction in cancer: a mechanism for immunosuppression. Immunol Cell Biol 83:451–461PubMedCrossRefGoogle Scholar
  21. 21.
    Preynat-Seauve O, Contassot E, Schuler P, Piguet V, French LE, Huard B (2007) Extralymphatic tumors prepare draining lymph nodes to invasion via a T-cell cross-tolerance process. Cancer Res 67:5009–5016PubMedCrossRefGoogle Scholar
  22. 22.
    Preynat-Seauve O, Schuler P, Contassot E, Beermann F, Huard B, French LE (2006) Tumor-infiltrating dendritic cells are potent antigen-presenting cells able to activate T cells and mediate tumor rejection. J Immunol 176:61–67PubMedGoogle Scholar
  23. 23.
    Robinson BW, Lake RA, Nelson DJ, Scott BA, Marzo AL (1999) Cross-presentation of tumour antigens: evaluation of threshold, duration, distribution and regulation. Immunol Cell Biol 77:552–558PubMedCrossRefGoogle Scholar
  24. 24.
    Tefany FJ, Barnetson RS, Halliday GM, McCarthy SW, McCarthy WH (1991) Immunocytochemical analysis of the cellular infiltrate in primary regressing and non-regressing malignant melanoma. J Invest Dermatol 97:197–202PubMedCrossRefGoogle Scholar
  25. 25.
    van Mierlo GJ, Boonman ZF, Dumortier HM, den Boer AT, Fransen MF, Nouta J, van der Voort EI, Offringa R, Toes RE, Melief CJ (2004) Activation of dendritic cells that cross-present tumor-derived antigen licenses CD8+ CTL to cause tumor eradication. J Immunol 173:6753–6759PubMedGoogle Scholar
  26. 26.
    Vicari AP, Chiodoni C, Vaure C, Ait-Yahia S, Dercamp C, Matsos F, Reynard O, Taverne C, Merle P, Colombo MP, O’Garra A, Trinchieri G, Caux C (2002) Reversal of tumor-induced dendritic cell paralysis by CpG immunostimulatory oligonucleotide and anti-interleukin 10 receptor antibody. J Exp Med 196:541–549PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2008

Authors and Affiliations

  • Patrizia Stoitzner
    • 1
    • 2
    Email author
  • Laura K. Green
    • 1
  • Jae Y. Jung
    • 1
  • Kylie M. Price
    • 1
  • Haley Atarea
    • 1
  • Bronwyn Kivell
    • 3
  • Franca Ronchese
    • 1
  1. 1.Malaghan Institute of Medical ResearchWellingtonNew Zealand
  2. 2.Department of Dermatology and VenereologyInnsbruck Medical UniversityInnsbruckAustria
  3. 3.Victoria UniversityWellingtonNew Zealand

Personalised recommendations