Skip to main content

Immunosuppressive activity of cancer-associated fibroblasts in head and neck squamous cell carcinoma

Cancer Immunology, Immunotherapy volume 64, pages 1407–1417 (2015)Cite this article

Abstract

Cancer-associated fibroblasts (CAFs) have been shown to play an important role in angiogenesis, invasion, and metastasis. In the present study, we determined whether CAFs within the tumor microenvironment (TME) in head and neck squamous cell carcinoma (HNSCC) contributed to promoting immunosuppression and evasion from immune surveillance. Six pairs of CAFs and normal fibroblasts (NFs) were established from the resected tumor tissues of patients with HNSCC. The effects of CAFs and NFs on the functions of T cells were comparatively analyzed. CAFs expressed the co-regulatory molecules, B7H1 and B7DC, whereas NFs did not. The expression levels of cytokine genes, including those for IL6, CXCL8, TNF, TGFB1, and VEGFA, were higher in CAFs. T cell proliferation was suppressed more by CAFs or their supernatants than by NFs. Moreover, PBMCs co-cultured with the supernatants of CAFs preferentially induced T cell apoptosis and regulatory T cells over those co-cultured with the supernatants of NFs. A microarray analysis revealed that the level of genes related to the leukocyte extravasation and paxillin signaling pathways was higher in CAFs than in NFs. These results demonstrated that CAFs collaborated with tumor cells in the TME to establish an immunosuppressive network that facilitated tumor evasion from immunological destruction.

This is a preview of subscription content, access via your institution.

Access options

Buy single article

Instant access to the full article PDF.

43,34 €

Price includes VAT (Finland)
Tax calculation will be finalised during checkout.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Abbreviations

7-ADD:

7-Amino-actinomycin D

APC:

Allophycocyanin

CAF:

Cancer-associated fibroblast

CCL7:

Chemokine ligand 7

CFSE:

Carboxyfluorescein succinimidyl ester

Cy:

Cyanine

FAP:

Fibroblast activation protein

GAPDH:

Glyceraldehyde-3-phosphate dehydrogenase

HGF:

Hepatocyte growth factor

HNSCC:

Head and neck squamous cell carcinoma

IGF:

Insulin-like growth factor

IL:

Interleukin

IPA:

Ingenuity pathway analysis

MMP:

Matrix metalloproteinase

NF:

Normal fibroblast

NSCLC:

Non-small cell lung cancer

SMA:

Smooth muscle actin

TGF:

Transforming growth factor

TME:

Tumor microenvironment

Treg:

Regulatory T cells

VEGF:

Vascular endothelial growth factor

References

  1. Siegel R, Naishandham D, Jemal A (2012) Cancer statistics, 2012. CA Cancer J Clin 62:10–29

    Article  PubMed  Google Scholar 

  2. Leemans CR, Braakhuis BJM, Brakenhoff RH (2011) The molecular biology of head and neck cancer. Nat Rev Cancer 11:9–22

    CAS  Article  PubMed  Google Scholar 

  3. Augsten M (2014) Cancer-associated fibroblasts as another polarized cell type of the tumor microenvironment. Front Oncol 4:62

    PubMed Central  Article  PubMed  Google Scholar 

  4. Franco OE, Shaw AK, Strand DW, Hayward SW (2010) Cancer associated fibroblasts in cancer pathogenesis. Semin Cell Dev Biol 21:33–39

    PubMed Central  CAS  Article  PubMed  Google Scholar 

  5. Kalluri R, Zeisberg M (2006) Fibroblasts in cancer. Nat Rev Cancer 6:392–401

    CAS  Article  PubMed  Google Scholar 

  6. Ito M, Ishii G, Nagai K, Maeda R, Nakano Y, Ochiai A (2012) Prognostic impact of cancer-associated stromal cells in patients with stage I lung adenocarcinoma. Chest 142:151–158

    Article  PubMed  Google Scholar 

  7. Herrera M, Herrera A, Domínguez G, Silva J, García V, García JM, Gómez I, Soldevilla B, Muñoz C, Provencio M, Campos-Martin Y, García de Herreros A et al (2013) Cancer-associated fibroblast and M2 macrophage markers together predict outcome in colorectal cancer patients. Cancer Sci 104:437–444

    CAS  Article  PubMed  Google Scholar 

  8. Zhang J, Liu J (2013) Tumor stroma as targets for cancer therapy. Pharmacol Ther 137:200–215

    PubMed Central  CAS  Article  PubMed  Google Scholar 

  9. Rasanen K, Vaheri A (2010) Activation of fibroblasts in cancer stroma. Exp Cell Res 316:2713–2722

    Article  PubMed  Google Scholar 

  10. Zhang L, Conejo-Garcia JR, Katsaros D, Gimotty PA, Massobrio M, Regnani G, Makrigiannakis A, Gray H, Schlienger K, Liebman MN, Rubin SC, Coukos G (2003) Intratumoral T cells, recurrence, and survival in epithelial ovarian cancer. N Engl J Med 348:203–213

    CAS  Article  PubMed  Google Scholar 

  11. Pages F, Berger A, Camus M, Sanchez-Cabo F, Costes A, Molidor R, Mlecnik B, Kirilovsky A, Nilsson M, Damotte D, Meatchi T, Bruneval P et al (2005) Effector memory T cells, early metastasis, and survival in colorectal cancer. N Engl J Med 353:2654–2666

    CAS  Article  PubMed  Google Scholar 

  12. Whiteside TL (2010) Immune responses to malignancies. J Allergy Clin Immunol 125:S272–S283

    PubMed Central  Article  PubMed  Google Scholar 

  13. Talmadge JE (2011) Immune cell infiltration of primary and metastatic lesions: mechanisms and clinical impact. Semin Cancer Biol 21:131–138

    CAS  Article  PubMed  Google Scholar 

  14. Poschke I, Mougiakakos D, Kiessling R (2011) Camouflage and sabotage: tumor escape from the immune system. Cancer Immunol Immunother 60:1161–1171

    CAS  Article  PubMed  Google Scholar 

  15. Hanahan D, Weinberg RA (2011) Hallmarks of cancer: the next generation. Cell 144:646–674

    CAS  Article  PubMed  Google Scholar 

  16. Tong CCL, Kao J, Silora AG (2012) Recognizing and reversing the immunosuppressive tumor microenvironment of head and neck cancer. Immunol Res 54:266–274

    CAS  Article  PubMed  Google Scholar 

  17. Balsamo M, Scordamaglia F, Pietra G, Manzini C, Cantoni C, Boitano M, Queirolo P, Vermi W, Facchetti F, Moretta A, Moretta L, Mingari MC et al (2009) Melanoma-associated fibroblasts modulate NK cell phenotype and antitumor cytotoxicity. Proc Natl Acad Sci USA 106:20847–20852

    PubMed Central  CAS  Article  PubMed  Google Scholar 

  18. De Monte L, Reni M, Tassi E, Clavenna D, Papa I, Recalde H, Braga M, Di Carlo V, Doglioni C, Protti MP (2011) Intratumor T helper type 2 cell infiltrate correlates with cancer-associated fibroblast thymic stromal lymphopoietin production and reduced survival in pancreatic cancer. J Exp Med 208:469–478

    PubMed Central  Article  PubMed  Google Scholar 

  19. Nazareth MR, Broderick L, Simpson-Abelson MR, Kelleher RJ Jr, Yokota SJ, Bankert RB (2007) Characterization of human lung tumor-associated fibroblasts and their ability to modulate the activation of tumor-associated T cells. J Immunol 178:5552–5562

    CAS  Article  PubMed  Google Scholar 

  20. Kyi C, Postow MA (2014) Checkpoint blocking antibodies in cancer immunotherapy. FEBS Lett 588:368–376

    CAS  Article  PubMed  Google Scholar 

  21. Hamid O, Carvajal RD (2013) Anti-programmed death-1 and anti-programmed death-ligand 1 antibodies in cancer therapy. Expert Opin Biol Ther 13:847–861

    CAS  Article  PubMed  Google Scholar 

  22. Brahmer JR, Tykodi SS, Chow LQ, Hwu WJ, Topalian SL, Hwu P, Drake CG, Camacho LH, Kauh J, Odunsi K, Pitot HC, Hamid O et al (2012) Safety and activity of anti-PD-L1 antibody in patients with advanced cancer. N Engl J Med 366:2455–2465

    PubMed Central  CAS  Article  PubMed  Google Scholar 

  23. Knowles LM, Stabile LP, Egloff AM, Rothstein ME, Thomas SM, Gubish CT, Lerner EC, Seethala RR, Suzuki S, Quesnelle KM, Morgan S, Ferris RL et al (2009) HGF and c-Met participate in paracrine tumorigenic pathways in head and neck squamous cell cancer. Clin Cancer Res 15:3740–3750

    PubMed Central  CAS  Article  PubMed  Google Scholar 

  24. Rosenthal E, McCrory A, Talbert M, Young G, Murphy-Ullrich J, Gladson C (2004) Elevated expression of TGF-β1 in head and neck cancer-associated fibroblasts. Mol Carcinog 40:116–121

    CAS  Article  PubMed  Google Scholar 

  25. Chen SF, Nieh S, Jao SW, Wu MZ, Liu CL, Chang YC, Lin YS (2013) The paracrine effect of cancer-associated fibroblast-induced interleukin-33 regulates the invasiveness of head and neck squamous cell carcinoma. J Pathol 231:180–189

    CAS  Article  PubMed  Google Scholar 

  26. Jung DW, Che ZM, Kim J, Kim K, Kim KY, Williams D, Kim J (2010) Tumor-stromal crosstalk in invasion of oral squamous cell carcinoma: a pivotal role of CCL7. Int J Cancer 127:332–344

    CAS  PubMed  Google Scholar 

  27. Johansson AC, Ansell A, Jerhammar F, Lindh MB, Grenman R, Munck-Wikland E, Östman A, Roberg K (2012) Cancer-associated fibroblasts induce matrix metalloproteinase-mediated cetuximab resistance in head and neck squamous cell carcinoma cells. Mol Cancer Res 10:1158–1168

    CAS  Article  PubMed  Google Scholar 

  28. Elkord E, Alcantar-Orozco EM, Dovedi SJ, Tran DQ, Hawkins RE, Gilham DE (2010) T regulatory cells in cancer: recent advances and therapeutic potential. Expert Opin Biol Ther 10:1573–1586

    CAS  Article  PubMed  Google Scholar 

  29. Knosp CA, Johnston JA (2012) Regulation of CD4+ T-cell polarization by suppressor of cytokine signalling proteins. Immunology 135:101–111

    PubMed Central  CAS  Article  PubMed  Google Scholar 

  30. Wada J, Suzuki H, Fuchino R, Yamasaki A, Nagai S, Yanai K, Koga K, Nakamura M, Tanaka M, Morisaki T, Katano M (2009) The contribution of vascular endothelial growth factor to the induction of regulatory T-cells in malignant effusions. Anticancer Res 29:881–888

    CAS  PubMed  Google Scholar 

  31. Mace TA, Ameen Z, Collins A, Wojcik S, Mair M, Young GS, Fuchs JR, Eubank TD, Frankel WL, Bekaii-Saab T, Bloomston M, Lesinski GB (2013) Pancreatic cancer-associated stellate cells promote differentiation of myeloid-derived suppressor cells in a STAT3-dependent manner. Cancer Res 73:3007–3018

    PubMed Central  CAS  Article  PubMed  Google Scholar 

  32. Gumbiner BM (1996) Cell adhesion: the molecular basis of tissue architecture and morphogenesis. Cell 84:345–357

    CAS  Article  PubMed  Google Scholar 

  33. Turner CE (2000) Paxillin and focal adhesion signaling. Nat Cell Biol 2:E231–E236

    CAS  Article  PubMed  Google Scholar 

Download references

Acknowledgments

This work was supported in part by KAKENHI (Grants-in-Aid for Scientific Research) (15K10798 to Koichi Sakakura, 25861525 to Minoru Toyoda, 26670736 to Kazuaki Chikamatsu). We thank Dr. Theresa L. Whiteside for her critical reading and comments.

Author information

Affiliations

  1. Department of Otolaryngology-Head and Neck Surgery, Gunma University Graduate School of Medicine, 3-39-22, Maebashi, Gunma, 371-8511, Japan

    Hideyuki Takahashi, Koichi Sakakura, Minoru Toyoda & Kazuaki Chikamatsu

  2. Division of Integrate Oncology Research, Gunma University Initiative for Advanced Research, 3-39-22, Maebashi, Gunma, 371-8511, Japan

    Reika Kawabata-Iwakawa

  3. Department of Molecular Pharmacology and Oncology, Gunma University Graduate School of Medicine, 3-39-22, Maebashi, Gunma, 371-8511, Japan

    Susumu Rokudai & Masahiko Nishiyama

Authors
  1. Hideyuki Takahashi
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Koichi Sakakura
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Reika Kawabata-Iwakawa
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Susumu Rokudai
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Minoru Toyoda
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Masahiko Nishiyama
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Kazuaki Chikamatsu
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Kazuaki Chikamatsu.

Ethics declarations

Conflict of interest

The authors declare that they have no conflict of interest.

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Takahashi, H., Sakakura, K., Kawabata-Iwakawa, R. et al. Immunosuppressive activity of cancer-associated fibroblasts in head and neck squamous cell carcinoma. Cancer Immunol Immunother 64, 1407–1417 (2015). https://doi.org/10.1007/s00262-015-1742-0

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00262-015-1742-0

Keywords

  • Cancer-associated fibroblasts
  • Head and neck squamous cell carcinoma
  • Immunosuppression
  • Regulatory T cells
  • T cell apoptosis