Arginase-1-based vaccination against the tumor microenvironment: the identification of an optimal T-cell epitope


l-arginine depletion by regulatory cells and cancer cells expressing arginase-1 (Arg-1) is a vital contributor to the immunosuppressive tumor microenvironment in patients with cancer. We have recently described the existence of pro-inflammatory effector T cells that recognize Arg-1. Hence, Arg-1-specific self-reactive T cells are a naturally occurring part of the memory T-cell repertoire of the human immune system. Here, we further characterize a highly immunogenic epitope from Arg-1. We describe frequent T-cell-based immune responses against this epitope in patients with cancer, as well as in healthy donors. Furthermore, we show that Arg-1-specific T cells expand in response to the TH2 cytokine interleukin (IL)-4 without any specific stimulation. Arg-1-specific memory TH1 cells that respond to increased IL-4 concentration may, therefore, drive the immune response back into the TH1 pathway. Arg-1-specific T cells thus appear to have an important function in immune regulation. Because Arg-1 plays an important role in the immunosuppressive microenvironment in most cancers, an immune modulatory vaccination approach can readily be employed to tilt the balance away from immune suppression in these settings.

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

Fig. 1
Fig. 2
Fig. 3
Fig. 4



Arginase 1


Breast cancer


Healthy donor


Malignant melanoma


Tumor-associated macrophages


Too numerous to count


  1. 1.

    Geiger R et al (2016) l-Arginine modulates T cell metabolism and enhances survival and anti-tumor activity. Cell 167(3):829–842.

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  2. 2.

    Zea AH et al (2004) L-Arginine modulates CD3ζ expression and T cell function in activated human T lymphocytes. Cell Immunol 232(1–2):21–31.

    CAS  Article  Google Scholar 

  3. 3.

    Rodriguez PC, Quiceno DG, Ochoa AC (2007) L-arginine availability regulates T-lymphocyte cell-cycle progression. Blood 109(4):1568–1574.

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  4. 4.

    De Boniface J, Mao Y, Schmidt-mende J, Kiessling R, Poschke I (2012) Expression patterns of the immunomodulatory enzyme arginase 1 in blood, lymph nodes and tumor tissue of early-stage breast cancer patients. Oncoimmunology 1(8):1305–1312.

    Article  PubMed  PubMed Central  Google Scholar 

  5. 5.

    Lang S et al (2018) Clinical relevance and suppressive capacity of human MDSC subsets. Clin Cancer Res 24(19):4834–4844.

    CAS  Article  Google Scholar 

  6. 6.

    Rodriguez PC et al (2009) Arginase I—producing myeloid-derived suppressor cells in renal cell carcinoma are a subpopulation of activated granulocytes. Cancer Res 69(4):1553–1561.

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  7. 7.

    Rotondo R et al (2009) IL-8 induces exocytosis of arginase 1 by neutrophil polymorphonuclears in nonsmall cell lung cancer. Int J Cancer 125:887–893.

    CAS  Article  Google Scholar 

  8. 8.

    Gajewski TF, Schreiber H, Fu YX (2013) Innate and adaptive immune cells in the tumor microenvironment. Nat Immunol 14(10):1014–1022.

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  9. 9.

    Andersen MH (2016) Anti-regulatory T cells. Semin Immunopathol.

    Article  Google Scholar 

  10. 10.

    Andersen MH (2015) Immune regulation by self-recognition: novel possibilities for anticancer immunotherapy. J Natl Cancer Inst 107(9):1–8.

    CAS  Article  Google Scholar 

  11. 11.

    Andersen MH (2018) The balance players of the adaptive immune system. Cancer Res 15:1379–1383.

    CAS  Article  Google Scholar 

  12. 12.

    Martinenaite E et al (2018) Frequent adaptive immune responses against arginase-1. Oncoimmunology 7(3):1–9.

    Article  Google Scholar 

  13. 13.

    Jørgensen MA et al (2018) Spontaneous T-cell responses against Arginase-1 in the chronic myeloproliferative neoplasms relative to disease stage and type of driver mutation. Oncoimmunology.

    Article  PubMed  PubMed Central  Google Scholar 

  14. 14.

    Martinenaite E, Ahmad SM, Svane IM, Andersen MH (2019) Peripheral memory T cells specific for Arginase-1. Cell Mol Immunol.

    Article  Google Scholar 

  15. 15.

    Moodie Z, Price L, Janetzki S, Cedrik B (2012) Response determination criteria for ELISPOT: toward a standard that can be applied across laboratories. Methods Mol Biol 792:185–196.

    CAS  Article  Google Scholar 

  16. 16.

    Cassetta L, Pollard JW (2018) Targeting macrophages: therapeutic approaches in cancer. Nat Rev Drug Discov 17(12):887–904.

    CAS  Article  Google Scholar 

  17. 17.

    Munder M, Eichmann K, Morán JM, Centeno F, Soler G, Modolell M (1999) Th1/Th2-regulated expression of arginase isoforms in murine macrophages and dendritic cells. J Immunol 163(7):3771–3777

    CAS  Google Scholar 

  18. 18.

    Yu W, Jiang N, Quake SR, Davis MM (2015) Clonal deletion prunes but does not eliminate self-specific alphabeta CD8(+) T lymphocytes. Immunity 42:929–941.

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  19. 19.

    Namdar A et al (2018) Prophylactic DNA vaccine targeting Foxp3 + regulatory T cells depletes myeloid-derived suppressor cells and improves anti-melanoma immune responses in a murine model. Cancer Immunol Immunother 67(3):367–379.

    CAS  Article  Google Scholar 

  20. 20.

    Nair S, Boczkowski D, Fassnacht M, Pisetsky D, Gilboa E (2007) Vaccination against the forkhead family transcription factor Foxp3 enhances tumor immunity. Cancer Res 67(1):371–380.

    CAS  Article  Google Scholar 

  21. 21.

    Munir S et al (2013) HLA-restricted CTL that are specific for the immune checkpoint ligand PD-L1 occur with high frequency in cancer patients. Cancer Res 73(6):1764–1776.

    CAS  Article  Google Scholar 

  22. 22.

    Munir S, Andersen GH, Svane IM, Andersen MH (2013) The immune checkpoint regulator PD-L1 is a specific target for naturally occurring CD4(+) T cells. Oncoimmunology 2(4):e23991.

    Article  PubMed  PubMed Central  Google Scholar 

  23. 23.

    Munir S, Andersen GH, Woetmann A, Ødum N, Becker JC, Andersen MH (2013) Cutaneous T cell lymphoma cells are targets for immune checkpoint ligand PD-L1-specific, cytotoxic T cells. Leukemia 27(11):2251–2253.

    CAS  Article  Google Scholar 

  24. 24.

    Ahmad SM, Larsen SK, Svane IM, Andersen MH (2014) Harnessing PD-L1-specific cytotoxic T cells for anti-leukemia immunotherapy to defeat mechanisms of immune escape mediated by the PD-1 pathway. Leukemia 28(1):236–238.

    CAS  Article  Google Scholar 

  25. 25.

    Soerensen RB, Hadrup SR, Svane IM, Hjortso MC, Straten PT, Andersen MH (2011) Indoleamine 2,3-dioxygenase specific, cytotoxic T cells as immune regulators. Blood 117(7):2200–2210.

    CAS  Article  Google Scholar 

  26. 26.

    Andersen MH (2012) CD4 responses against IDO. Oncoimmunology 1(7):1211–1212.

    Article  PubMed  PubMed Central  Google Scholar 

  27. 27.

    Larsen SK et al (2013) Functional characterization of Foxp3-specific spontaneous immune responses. Leukemia 27(12):2332–2340.

    CAS  Article  Google Scholar 

  28. 28.

    Martinenaite E et al (2016) CCL22-specific T Cells: modulating the immunosuppressive tumor microenvironment. Oncoimmunology 5(11):e1238541.

    CAS  Article  PubMed  PubMed Central  Google Scholar 

Download references


This work has been supported by grants from the Danish Cancer Society (Grant number R146-A9440-16-S2), Herlev Hospital (CCIT-Dk funding) and Innovation Fund Denmark (Grant number 8054-00058B).

Author information




MHA designed and supervised the study. EM designed the experiments and analyzed the data. EM, SMA, SKB, MAJ, and SEWB performed experiments. IMS provided the relevant clinical material. All authors contributed to drafting the manuscript.

Corresponding author

Correspondence to Mads Hald Andersen.

Ethics declarations

Conflict of interest

Mads Hald Andersen has filed several patent applications based on the use of arginase for vaccinations. The rights of the patent applications have been transferred to Copenhagen University Hospital, Herlev, in accordance with the Danish Law of Public Inventions at Public Research Institutions. The capital region has licensed these patents to the company IO Biotech ApS, whose purpose is to develop immune-modulating vaccines for cancer treatments. Mads Hald Andersen is a shareholder and board member of IO Biotech ApS. Evelina Martinenaite is employed by IO Biotech ApS. Other authors declare no conflict of interest.

Ethical approval and ethical standards

The protocol was approved by the Scientific Ethics Committee for the Capital Region of Denmark (H-A-2009-013) and conducted in accordance with the provisions of the Declaration of Helsinki.

Informed consent

Written informed consent for the use of the PBMCs for research purposes was obtained from the patients and healthy donors prior to inclusion in the study.

Additional information

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

This paper is a Focussed Research Review based on a presentation given at the Eighteenth International Conference on Progress in Vaccination against Cancer (PIVAC 18), held in Oslo, Norway, 3rd–5th October, 2018. It is part of a Cancer Immunology, Immunotherapy series of PIVAC 18 papers.

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Martinenaite, E., Ahmad, S.M., Bendtsen, S.K. et al. Arginase-1-based vaccination against the tumor microenvironment: the identification of an optimal T-cell epitope. Cancer Immunol Immunother 68, 1901–1907 (2019).

Download citation


  • Arginase
  • Anti-regulatory T cells
  • Immune-modulating vaccines
  • IO112
  • PIVAC 18