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Circulating T regulatory cells migration and phenotype in glioblastoma patients: an in vitro study

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Abstract

Glioblastoma multiforme (GBM) is the most aggressive primary human brain tumor. The relatively high amount of T regulatory lymphocytes present in the tumor, contributes to the establishment of an immunosuppressive microenvironment. Samples of peripheral blood were collected from GBM patients and healthy controls and a purified population of Treg (CD4+/CD25bright) was isolated using flow cytometric cell sorting. Treg migrating capacities toward human glioma cell line conditioned medium were evaluated through an in vitro migration test. Our data show that supernatants collected from GBM cell lines were more attractant to Treg when compared to complete standard medium. The addition of an anti-CCL2 antibody to conditioned medium decreased conditioned medium-depending Treg migration, suggesting that CCL2 (also known as Monocyte Chemoattractant Protein, MCP-1) is implicated in the process. The number of circulating CD4+/μL or Treg/μL was similar in GBM patients and controls. Specific Treg markers (FOXP3; CD127; Helios; GITR; CTLA4; CD95; CCR2, CCR4; CCR7) were screened in peripheral blood and no differences could be detected between the two populations. These data confirm that the tumor microenvironment is attractive to Treg, which tend to migrate toward the tumor region changing the immunological response. Though we provide evidence that CCL2 is implicated in Treg migration, other factors are needed as well to provide such effect.

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References

  1. Wainwright DA, Nigam P, Thaci B et al (2012) Recent developments on immunotherapy for brain cancer. Expert Opin Emerg Drugs 17(2):181–202

    Article  PubMed  CAS  Google Scholar 

  2. Behin A, Hoang-Xuan K, Carpentier AF et al (2003) Primary brain tumors in adults. Lancet 361:323–331

    Article  PubMed  Google Scholar 

  3. Curtin JF, Candolfi M, Fakhouri TM et al (2008) Treg depletion inhibits efficacy of cancer immunotherapy: implications for clinical trials. PLoS ONE 3(4):e1983

    Article  PubMed  Google Scholar 

  4. Crane CA, Ahn BJ, Han SJ et al (2012) Soluble factors secreted by glioblastoma cell lines facilitate recruitment, survival, and expansion of regulatory T cells: implications for immunotherapy. Neuro Oncol 14(5):584–595

    Article  PubMed  CAS  Google Scholar 

  5. Fujimoto S, Greene MI, Sehon AH (1976) Regulation of the immuneresponse to tumor antigens. II. The nature of immunosuppressor cells in tumor-bearing hosts. J Immunol 116:800–806

    PubMed  CAS  Google Scholar 

  6. Ha TY, Park HJ, On KK et al (1995) Potentiation of anti-tumor immunity by modulation of suppressor T cells. II. Regression of tumor by modulation of suppressor T cells. Korean J Immunol 17:101–118

    Google Scholar 

  7. Ha TY (2009) The role of regulatory T cells in cancer. Immune Netw 9(6):209–235

    Article  PubMed  Google Scholar 

  8. Chen X, Oppenheim JJ (2011) Resolving the identity myth: key markers of functional CD4+ FoxP3+ regulatory T cells. Int Immunopharmacol 11(10):1489–1496

    Article  PubMed  CAS  Google Scholar 

  9. Wang RF (2006) Regulatory T cells and innate immune regulation in tumor immunity. Springer Semin Imunopathol 28:17–23

    Article  Google Scholar 

  10. Fietta AM, Morosini M, Passadore I et al (2009) Systemic inflammatory response and downmodulation of peripheral CD25+ Foxp3+ T-regulatory cells in patients undergoing radiofrequency thermal ablation for lung cancer. Human Immunol 70:477–486

    Article  CAS  Google Scholar 

  11. Liu Z, Kim JH, Falo LD et al (2009) Tumor regulatory T cells potently abrogate antitimor immunity. J Immunol 182:6160–6167

    Article  PubMed  CAS  Google Scholar 

  12. El Andaloussi A, Lesniak MS (2006) An increase in CD4+ CD25+ FOXP3+ regulatory T cells in tumor infiltrating lymphocytes of human glioblastoma multiforme. Neuro Oncol 8:234–243

    Article  PubMed  Google Scholar 

  13. Heimberger AB, Abou-Ghazal M, Reina-Ortiz C et al (2008) Incidence and prognostic impact of FoxP3+ regulatory T cells in human gliomas. Clin Cancer Res 14:5166–5172

    Article  PubMed  CAS  Google Scholar 

  14. Jacobs JF, Idema AJ, Bol KF et al (2010) Prognostic significance and mechanism of Treg infiltration in human brain tumors. J Neuroimmunol 225:195–199

    Article  PubMed  CAS  Google Scholar 

  15. Fecci PE, Mitchell DA, Whitesides JF et al (2006) Increased regulatory T-cell fraction amidst a diminished CD4 compartment explains cellular immune defects in patients with malignant glioma. Cancer Res 66(6):3294–3302

    Article  PubMed  CAS  Google Scholar 

  16. Zhang J, Patel L, Pienta KJ (2010) Targeting chemokine (C–C motif) ligand 2 (CCL2) as an example of translation of cancer molecular biology to the clinic. Prog Mol Biol Transl Sci 95:31–53

    Article  PubMed  CAS  Google Scholar 

  17. Deshmane SL, Kremlev S, Amini S et al (2009) Monocyte chemoattractant protein-1 (MCP-1): an overview. J Interferon Cytokine Res 29:313–326

    Article  PubMed  CAS  Google Scholar 

  18. Del Pozo MA, Cabanas C, Montoya MC et al (1997) ICAMs redistributed by chemokines to cellular uropods as a mechanism for recruitment of T lymphocytes. J Cell Biol 137:493–508

    Article  PubMed  Google Scholar 

  19. Carrillo-de Sauvage MA, Gómez A, Ros CM (2012) CCL2-expressing astrocytes mediate the extravasation of T lymphocytes in the brain. Evidence from patients with glioma and experimental models in vivo. PLoS ONE 7:30762

    Article  Google Scholar 

  20. Calatozzolo C, Canazza A, Pollo B et al (2011) Expression of the new CXCL12 receptor, CXCR7, in gliomas. Cancer Biol Ther 11(242–53):17

    Google Scholar 

  21. Conductier G, Blondeau N, Guyon A et al (2010) The role of monocyte chemoattractant protein CCL2/CCL2 in neuroinflammatory diseases. J Neuroimmunol 224:93–100

    Article  PubMed  CAS  Google Scholar 

  22. Kim YH, Jung TY, Jung S et al (2012) Tumor-infiltrating T-cell subpopulations in glioblastomas. Br J Neurosurg 26:21–27

    Article  PubMed  Google Scholar 

  23. Donson AM, Birks DK, Schittone SA et al (2012) Increased immune gene expression and immune cell infiltration in high grade astrocytoma distinguish long-term from short-term survivors. J Immunol 189:1920–1927

    Article  PubMed  CAS  Google Scholar 

  24. Lowther DE, Hafler DA (2012) Regulatory T cells in the central nervous system. Immunol Rev 248(1):156–169

    Article  PubMed  Google Scholar 

  25. Jordan JT, Sun W, Hussain SF et al (2008) Preferential migration of regulatory T cells mediated by glioma-secreted chemokines can be blocked with chemotherapy. Cancer Immunol Immunother 57(1):123–131

    Article  PubMed  CAS  Google Scholar 

  26. Parney IF, Waldron J, Parsa AT (2009) Flow citometry and in vitro analysis of human glioma associated macrophages. J Neurosurg 110(3):572–582

    Article  PubMed  CAS  Google Scholar 

  27. Lohr J, Ratliff T, Huppertz A et al (2011) Effector T-cell infiltration positively impacts survival of glioblastoma patients and is impaired by tumor-derived TGF-β. Clin Cancer Res 17(13):4296–4308

    Article  PubMed  CAS  Google Scholar 

  28. de Chaisemartin L, Goc J, Damotte D et al (2011) Characterization of chemokines and adhesion molecules associated with T cell presence in tertiary lymphoid structures in human lung cancer. Cancer Res 71(20):6391–6399

    Article  PubMed  Google Scholar 

  29. Wei S, Kryczek I, Zou W (2006) Regulatory T-cell compartmentalization and trafficking. Blood 108(2):426–431

    Article  PubMed  CAS  Google Scholar 

  30. Ménétrier-Caux C, Faget J, Biota C, Gobert M, Blay JY, Caux C (2012) Innate immune recognition of breast tumor cells mediates CCL2 secretion favoring Treg recruitment within tumorenvironment. Oncoimmunology 1(5):759–761

    Article  PubMed  Google Scholar 

  31. Weller M (2012) More on the immune privilege of glioblastoma. Neuro Oncol 14(5):527–528

    Article  PubMed  Google Scholar 

  32. Nalla AK, Gogineni VR, Gupta R et al (2011) Suppression of uPA and uPAR blocks radiation-induced CCL2 mediated recruitment of endothelial cells in meningioma. Cell Signal 23(8):1299–1310

    Article  PubMed  CAS  Google Scholar 

  33. Yan M, Jene N, Byrne D et al (2011) Recruitment of regulatory T cells is correlated with hypoxia-induced CXCR4 expression, and is associated with poor prognosis in basal-like breast cancers. Breast Cancer Res 13(2):R47

    Article  PubMed  CAS  Google Scholar 

  34. Wainwright DA, Balyasnikova IV, Chang AL et al (2012) IDO Expression in brain tumors increases the recruitment of regulatory T cells and negatively impacts survival. Clin Cancer Res 18(22):6110–6121

    Article  PubMed  CAS  Google Scholar 

  35. Coe D, Begom S, Addey C et al (2010) Depletion of regulatory T cells by anti-GITR mAb as a novel mechanism for cancer immunotherapy. Cancer Immunol Immunother 59:1367–1377

    Article  PubMed  CAS  Google Scholar 

  36. Cohen AD, Schaer DA, Liu C et al (2010) Agonist anti-GITR monoclonal antibody induces melanoma tumor immunity in mice by altering regulatory T cell stability and intra-tumor accumulation. PLoS ONE 5:e10436

    Article  PubMed  Google Scholar 

  37. Faget J, Bendriss-Vermare N, Gobert M et al (2012) ICOS-ligand expression on plasmacytoid dendritic cells supports breast cancer progression by promoting the accumulation of immunosuppressive CD4+ T cells. Cancer Res 72:6130–6141

    Article  PubMed  CAS  Google Scholar 

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Acknowledgments

This work was supported by research Grant of the Ministero della Salute (LR9 IRCCS Foundation Neurological Institute “C. Besta”, Milan Italy). The authors are grateful to Luisa Fumagalli and Francesco Ghielmetti from Radiotherapy Unit (Foundation IRCCS Neurological Institute C. Besta) for the skillful technical help with irradiation experiments. We are also thankful to Marta Dossena from Cellular Neurobiology Laboratory, Cerebrovascular Diseases Unit (Foundation IRCCS Neurological Institute C. Besta) for her contribution on data analysis.

Conflict of interest

The authors declare that they have no conflict of interest.

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Author notes

  1. Alessandra Canazza

    Present address: Cellular Neurobiology Laboratory, Cerebrovascular Diseases Unit, Foundation IRCCS Neurological Institute C. Besta, Via Celoria, 11, 20133, Milan, Italy

Authors and Affiliations

  1. Laboratory of Clinical Pathology and Medical Genetics, Foundation IRCCS Neurological Institute C. Besta, Via Celoria, 11, 20133, Milan, Italy

    Chiara Vasco, Alessandra Canazza, Ambra Rizzo, Elena Corsini & Emilio Ciusani

  2. Department of Neuroncology, Foundation IRCCS Neurological Institute C. Besta, Via Celoria, 11, 20133, Milan, Italy

    Adele Mossa & Antonio Silvani

  3. Department of Radiotherapy, Foundation IRCCS Neurological Institute C. Besta, Via Celoria, 11, 20133, Milan, Italy

    Laura Fariselli

  4. Neurologia-Stroke Unit, Manzoni Hospital, Via Dell’Eremo 9/11, 23900, Lecco, Italy

    Andrea Salmaggi

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  1. Chiara Vasco
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Corresponding author

Correspondence to Emilio Ciusani.

Additional information

Chiara Vasco and Alessandra Canazza have equally contributed to this work.

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Vasco, C., Canazza, A., Rizzo, A. et al. Circulating T regulatory cells migration and phenotype in glioblastoma patients: an in vitro study. J Neurooncol 115, 353–363 (2013). https://doi.org/10.1007/s11060-013-1236-x

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  • DOI: https://doi.org/10.1007/s11060-013-1236-x

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