Desmoid tumors display a strong immune infiltration at the tumor margins and no PD-L1-driven immune suppression

  • Vasiliki SiozopoulouEmail author
  • Elly Marcq
  • Julie Jacobs
  • Karen Zwaenepoel
  • Christophe Hermans
  • Jantine Brauns
  • Siegrid Pauwels
  • Clément Huysentruyt
  • Martin Lammens
  • Johan Somville
  • Evelien Smits
  • Patrick Pauwels
Original Article


Desmoid tumors (DTs) are local aggressive neoplasms, whose therapeutic approach has remained so far unsolved and in many instances controversial. Nowadays, immunotherapy appears to play a leading role in the treatment of various tumor types. Characterization of the tumor immune microenvironment (TME) and immune checkpoints can possibly help identify new immunotherapeutic targets for DTs. We performed immunohistochemistry (IHC) on 33 formalin-fixed paraffin-embedded (FFPE) tissue sections from DT samples to characterize the TME and the immune checkpoint expression profile. We stained for CD3, CD4, CD8, CD20, FoxP3, CD45RO, CD56, CD68, NKp46, granzyme B, CD27, CD70, PD1 and PD-L1. We investigated the expression of the markers in the tumoral stroma, as well as at the periphery of the tumor. We found that most of the tumors showed organization of lymphocytes into lymphoid aggregates at the periphery of the tumor, strongly resembling tertiary lymphoid organs (TLOs). The tumor expressed a significant number of memory T cells, both at the periphery and in the tumoral stroma. In the lymphoid aggregates, we also recognized a significant proportion of regulatory T cells. The immune checkpoint ligand PD-L1 was negative on the tumor cells in almost all samples. On the other hand, PD1 was partially expressed in lymphocytes at the periphery of the tumor. To conclude, we are the first to show that DTs display a strong immune infiltration at the tumor margins, with formation of lymphoid aggregates. Moreover, we demonstrated that there is no PD-L1-driven immune suppression present in the tumor cells.


Desmoid tumors PD-L1 Immunotherapy Immunohistochemistry 



Cytotoxic T lymphocyte antigen-4


Desmoid tumors


Familial adenomatous polyposis


Formalin-fixed paraffin embedded


High endothelial venules


High-resolution melting analysis




Lymph node


Next-generation sequencing


Programmed death (ligand)-1




Regulatory T cells


Tertiary lymphoid organs


Tumor microenvironment



The majority of human biological material used in this publication was provided by the Tumor bank, Antwerp University Hospital, Belgium, which is funded by the National Cancer Plan.

Author contributions

VS, EM and JJ designed the study and performed the data acquisition and analysis. CH processed the slides. KZ and SP performed the immunohistochemical staining. CH provided patient material. All authors contributed to the interpretation of the data, sample collection, drafting and revision of the manuscript.


The authors received no specific funding for this work.

Compliance with ethical standards

Conflict of interest

All authors declare that they have no conflicts of interest.

Ethical approval and ethical standards

We received approval by the Ethics Committee of the Antwerp University Hospital/University of Antwerp (EC 18/45/517) to use historical samples. As it was a retrospective study, no informed consent of the patients could be obtained.

Supplementary material

262_2019_2390_MOESM1_ESM.pdf (314 kb)
Supplementary material 1 (PDF 314 kb)


  1. 1.
    Rizvi NA, Mazieres J, Planchard D et al (2015) Activity and safety of nivolumab, an anti-PD-1 immune checkpoint inhibitor, for patients with advanced, refractory squamous non-small-cell lung cancer (CheckMate 063): a phase 2, single-arm trial. Lancet Oncol 16:257–265. CrossRefGoogle Scholar
  2. 2.
    Escobar C, Munker R, Thomas JO, Li BD, Burton GV (2012) Update on desmoid tumors. Ann Oncol 23:562–569. CrossRefGoogle Scholar
  3. 3.
    Lazar AJ, Tuvin D, Hajibashi S et al (2008) Specific mutations in the beta-catenin gene (CTNNB1) correlate with local recurrence in sporadic desmoid tumors. Am J Pathol 173:1518–1527. CrossRefGoogle Scholar
  4. 4.
    Gurbuz AK, Giardiello FM, Petersen GM, Krush AJ, Offerhaus GJ, Booker SV, Kerr MC, Hamilton SR (1994) Desmoid tumours in familial adenomatous polyposis. Gut 35:377–381CrossRefGoogle Scholar
  5. 5.
    Calvert GT, Monument MJ, Burt RW, Jones KB, Randall RL (2012) Extra-abdominal desmoid tumors associated with familial adenomatous polyposis. Sarcoma 2012:726537. Google Scholar
  6. 6.
    Carlson JW, Fletcher CD (2007) Immunohistochemistry for beta-catenin in the differential diagnosis of spindle cell lesions: analysis of a series and review of the literature. Histopathology 51:509–514. CrossRefGoogle Scholar
  7. 7.
    Kasper B, Strobel P, Hohenberger P (2011) Desmoid tumors: clinical features and treatment options for advanced disease. Oncologist 16:682–693. CrossRefGoogle Scholar
  8. 8.
    Kasper B, Baumgarten C, Bonvalot S et al (2015) Management of sporadic desmoid-type fibromatosis: a European consensus approach based on patients’ and professionals’ expertise—a sarcoma patients EuroNet and European organisation for research and treatment of cancer/soft tissue and bone sarcoma group initiative. Eur J Cancer 51:127–136. CrossRefGoogle Scholar
  9. 9.
    Mahoney KM, Rennert PD, Freeman GJ (2015) Combination cancer immunotherapy and new immunomodulatory targets. Nat Rev Drug Discov 14:561–584. CrossRefGoogle Scholar
  10. 10.
    Sharma P, Allison JP (2015) The future of immune checkpoint therapy. Science 348:56–61. CrossRefGoogle Scholar
  11. 11.
    Burgess M, Tawbi H (2015) Immunotherapeutic approaches to sarcoma. Curr Treat Options Oncol 16:26. CrossRefGoogle Scholar
  12. 12.
    Jacobs J, Zwaenepoel K, Rolfo C et al (2015) Unlocking the potential of CD70 as a novel immunotherapeutic target for non-small cell lung cancer. Oncotarget 6:13462–13475. CrossRefGoogle Scholar
  13. 13.
    Marcq E, Siozopoulou V, De Waele J et al (2017) Prognostic and predictive aspects of the tumor immune microenvironment and immune checkpoints in malignant pleural mesothelioma. Oncoimmunology 6:e1261241. CrossRefGoogle Scholar
  14. 14.
    Ager A (2017) High endothelial venules and other blood vessels: critical regulators of lymphoid organ development and function. Front Immunol 8:45. CrossRefGoogle Scholar
  15. 15.
    Martinet L, Le Guellec S, Filleron T, Lamant L, Meyer N, Rochaix P, Garrido I, Girard JP (2012) High endothelial venules (HEVs) in human melanoma lesions: major gateways for tumor-infiltrating lymphocytes. Oncoimmunology 1:829–839. CrossRefGoogle Scholar
  16. 16.
    Martinet L, Garrido I, Filleron T, Le Guellec S, Bellard E, Fournie JJ, Rochaix P, Girard JP (2011) Human solid tumors contain high endothelial venules: association with T- and B-lymphocyte infiltration and favorable prognosis in breast cancer. Cancer Res 71:5678–5687. CrossRefGoogle Scholar
  17. 17.
    Jones GW, Hill DG, Jones SA (2016) Understanding immune cells in tertiary lymphoid organ development: it is all starting to come together. Front Immunol 7:401. CrossRefGoogle Scholar
  18. 18.
    Jing F, Choi EY (2016) Potential of cells and cytokines/chemokines to regulate tertiary lymphoid structures in human diseases. Immune Netw 16:271–280. CrossRefGoogle Scholar
  19. 19.
    Dieu-Nosjean MC, Antoine M, Danel C et al (2008) Long-term survival for patients with non-small-cell lung cancer with intratumoral lymphoid structures. J Clin Oncol 26:4410–4417. CrossRefGoogle Scholar
  20. 20.
    Pages F, Galon J, Dieu-Nosjean MC, Tartour E, Sautes-Fridman C, Fridman WH (2010) Immune infiltration in human tumors: a prognostic factor that should not be ignored. Oncogene 29:1093–1102. CrossRefGoogle Scholar
  21. 21.
    Hori S, Nomura T, Sakaguchi S (2003) Control of regulatory T cell development by the transcription factor Foxp3. Science 299:1057–1061. CrossRefGoogle Scholar
  22. 22.
    Hinz S, Pagerols-Raluy L, Oberg HH et al (2007) Foxp3 expression in pancreatic carcinoma cells as a novel mechanism of immune evasion in cancer. Cancer Res 67:8344–8350. CrossRefGoogle Scholar
  23. 23.
    Chanmee T, Ontong P, Konno K, Itano N (2014) Tumor-associated macrophages as major players in the tumor microenvironment. Cancers (Basel) 6:1670–1690. CrossRefGoogle Scholar
  24. 24.
    Jinushi M, Komohara Y (2015) Tumor-associated macrophages as an emerging target against tumors: creating a new path from bench to bedside. Biochim Biophys Acta 1855:123–130. Google Scholar
  25. 25.
    Curiel TJ, Coukos G, Zou L et al (2004) Specific recruitment of regulatory T cells in ovarian carcinoma fosters immune privilege and predicts reduced survival. Nat Med 10:942–949. CrossRefGoogle Scholar
  26. 26.
    Mizukami Y, Kono K, Kawaguchi Y, Akaike H, Kamimura K, Sugai H, Fujii H (2008) CCL17 and CCL22 chemokines within tumor microenvironment are related to accumulation of Foxp3 + regulatory T cells in gastric cancer. Int J Cancer 122:2286–2293. CrossRefGoogle Scholar
  27. 27.
    Matter M, Mumprecht S, Pinschewer DD, Pavelic V, Yagita H, Krautwald S, Borst J, Ochsenbein AF (2005) Virus-induced polyclonal B cell activation improves protective CTL memory via retained CD27 expression on memory CTL. Eur J Immunol 35:3229–3239. CrossRefGoogle Scholar
  28. 28.
    Claus C, Riether C, Schurch C, Matter MS, Hilmenyuk T, Ochsenbein AF (2012) CD27 signaling increases the frequency of regulatory T cells and promotes tumor growth. Cancer Res 72:3664–3676. CrossRefGoogle Scholar
  29. 29.
    Matter MS, Claus C, Ochsenbein AF (2008) CD4 + T cell help improves CD8 + T cell memory by retained CD27 expression. Eur J Immunol 38:1847–1856. CrossRefGoogle Scholar
  30. 30.
    Jiang Y, Li Y, Zhu B (2015) T-cell exhaustion in the tumor microenvironment. Cell Death Dis 6:e1792. CrossRefGoogle Scholar
  31. 31.
    Ruprecht CR, Gattorno M, Ferlito F, Gregorio A, Martini A, Lanzavecchia A, Sallusto F (2005) Coexpression of CD25 and CD27 identifies FoxP3 + regulatory T cells in inflamed synovia. J Exp Med 201:1793–1803. CrossRefGoogle Scholar
  32. 32.
    Duggleby RC, Shaw TN, Jarvis LB, Kaur G, Gaston JS (2007) CD27 expression discriminates between regulatory and non-regulatory cells after expansion of human peripheral blood CD4 + CD25 + cells. Immunology 121:129–139. CrossRefGoogle Scholar
  33. 33.
    Mlecnik B, Tosolini M, Kirilovsky A et al (2011) Histopathologic-based prognostic factors of colorectal cancers are associated with the state of the local immune reaction. J Clin Oncol 29:610–618. CrossRefGoogle Scholar
  34. 34.
    Hadrup S, Donia M, Thor Straten P (2013) Effector CD4 and CD8 T cells and their role in the tumor microenvironment. Cancer Microenviron 6:123–133. CrossRefGoogle Scholar
  35. 35.
    Galon J, Costes A, Sanchez-Cabo F et al (2006) Type, density, and location of immune cells within human colorectal tumors predict clinical outcome. Science 313:1960–1964. CrossRefGoogle Scholar
  36. 36.
    Galon J, Pages F, Marincola FM, Thurin M, Trinchieri G, Fox BA, Gajewski TF, Ascierto PA (2012) The immune score as a new possible approach for the classification of cancer. J Transl Med 10:1. CrossRefGoogle Scholar
  37. 37.
    Fridman WH, Pages F, Sautes-Fridman C, Galon J (2012) The immune contexture in human tumours: impact on clinical outcome. Nat Rev Cancer 12:298–306. CrossRefGoogle Scholar
  38. 38.
    Giraldo NA, Becht E, Pages F et al (2015) Orchestration and prognostic significance of immune checkpoints in the microenvironment of primary and metastatic renal cell cancer. Clin Cancer Res 21:3031–3040. CrossRefGoogle Scholar
  39. 39.
    Appay V, van Lier RA, Sallusto F, Roederer M (2008) Phenotype and function of human T lymphocyte subsets: consensus and issues. Cytometry A 73:975–983. CrossRefGoogle Scholar
  40. 40.
    Donnem T, Hald SM, Paulsen EE et al (2015) Stromal CD8 + T-cell density-A promising supplement to TNM staging in non-small cell lung cancer. Clin Cancer Res 21:2635–2643. CrossRefGoogle Scholar
  41. 41.
    Mandelboim O, Porgador A (2001) NKp46. Int J Biochem Cell Biol 33:1147–1150CrossRefGoogle Scholar
  42. 42.
    Prendergast GC, Jaffee EM (2013) Cancer immunotherapy: immune suppression and tumor growth, 2nd edition. Elsevier/AP, Academic Press is an imprint of Elsevier, Amsterdam; BostonGoogle Scholar
  43. 43.
    Riazi Rad F, Ajdary S, Omranipour R, Alimohammadian MH, Hassan ZM (2015) Comparative analysis of CD4 + and CD8 + T cells in tumor tissues, lymph nodes and the peripheral blood from patients with breast cancer. Iran Biomed J 19:35–44Google Scholar
  44. 44.
    Chen DS, Mellman I (2013) Oncology meets immunology: the cancer-immunity cycle. Immunity 39:1–10. CrossRefGoogle Scholar
  45. 45.
    Lee J, Ahn E, Kissick HT, Ahmed R (2015) Reinvigorating exhausted T cells by blockade of the PD-1 pathway. For Immunopathol Dis Therap 6:7–17. Google Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  • Vasiliki Siozopoulou
    • 1
    • 2
    Email author
  • Elly Marcq
    • 1
  • Julie Jacobs
    • 1
    • 2
  • Karen Zwaenepoel
    • 1
    • 2
  • Christophe Hermans
    • 1
    • 2
  • Jantine Brauns
    • 1
    • 2
  • Siegrid Pauwels
    • 2
  • Clément Huysentruyt
    • 3
  • Martin Lammens
    • 2
  • Johan Somville
    • 4
  • Evelien Smits
    • 1
    • 5
  • Patrick Pauwels
    • 1
    • 2
  1. 1.Center for Oncological ResearchUniversity of AntwerpAntwerpBelgium
  2. 2.Department of PathologyAntwerp University HospitalAntwerpBelgium
  3. 3.PAMM Laboratory for Pathology and Medical MicrobiologyCatharina HospitalEindhovenThe Netherlands
  4. 4.Department of Orthopedic SurgeryAntwerp University HospitalAntwerpBelgium
  5. 5.Center for Cell Therapy and Regenerative MedicineAntwerp University HospitalAntwerpBelgium

Personalised recommendations