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Predominance of M2 macrophages in gliomas leads to the suppression of local and systemic immunity

Abstract

Glioblastoma is a highly prevalent and aggressive form of primary brain tumor. It represents approximately 56% of all the newly diagnosed gliomas. Macrophages are one of the major constituents of tumor-infiltrating immune cells in the human gliomas. The role of immunosuppressive macrophages is very well documented in correlation with the poor prognosis of patients suffering from breast, prostate, bladder and cervical cancers. The current study highlights the correlation between the tumor-associated macrophage phenotypes and glioma progression. We observed an increase in the pool of M2 macrophages in high-grade gliomas, as confirmed by their CD68 and CD163 double-positive phenotype. In contrast, less M1 macrophages were noticed in high-grade gliomas, as evidenced by the down-regulation in the expression of CCL3 marker. In addition, we observed that higher gene expression ratio of CD163/CCL3 is associated with glioma progression. The Kaplan–Meier survival plots indicate that glioma patients with lower expression of M2c marker (CD163), and higher expression of M1 marker (CCL3) had better survival. Furthermore, we examined the systemic immune response in the peripheral blood and noted a predominance of M2 macrophages, myeloid-derived suppressor cells and PD-1+ CD4 T cells in glioma patients. Thus, the study indicates a high gene expression ratio of CD163/CCL3 in high-grade gliomas as compared to low-grade gliomas and significantly elevated frequency of M2 macrophages and PD-1+ CD4 T cells in the blood of tumor patients. These parameters could be used as an indicator of the early diagnosis and prognosis of the disease.

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Abbreviations

Abs:

Antibodies

DAB:

(3,3′-diaminobenzidine)

FACS:

Fluorescence-activated cell sorting

GAPDH:

Glyceraldehyde 3-phosphate dehydrogenase

HGG:

High-grade glioma

HLA-DR:

Human Leukocyte Antigen—DR isotype

IFN-γ:

Interferon gamma

IL-10:

Interleukin 10

IL-10R:

Interleukin-10 receptor

LGG:

Low-grade glioma

LPS:

Lipopolysaccharides

PBMCs:

Peripheral blood mononuclear cells

PBS:

Phosphate-buffered saline

PD-1:

Programmed cell death protein 1

TAM:

Tumor-associated macrophages

TCGA:

The Cancer Genome Atlas

TGF-β:

Transforming growth factor-beta

References

  1. 1.

    Broekman ML, Maas SLN, Abels ER, Mempel TR, Krichevsky AM, Breakefield XO (2018) Multidimensional communication in the microenvirons of glioblastoma. Nat Rev Neurol. https://doi.org/10.1038/s41582-018-0025-8

    Article  PubMed  PubMed Central  Google Scholar 

  2. 2.

    Preusser M, de Ribaupierre S, Wohrer A, Erridge SC, Hegi M, Weller M, Stupp R (2011) Current concepts and management of glioblastoma. Ann Neurol 70:9–21. https://doi.org/10.1002/ana.22425

    Article  PubMed  Google Scholar 

  3. 3.

    Preusser M, Lim M, Hafler DA, Reardon DA, Sampson JH (2015) Prospects of immune checkpoint modulators in the treatment of glioblastoma. Nat Rev Neurol 11:504–514. https://doi.org/10.1038/nrneurol.2015.139

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  4. 4.

    Arevalo AST, Erices JI, Uribe DA, Howden J, Niechi I, Munoz S, Martin RS, Monras CAQ (2017) Current therapeutic alternatives and new perspectives in glioblastoma multiforme. Curr Med Chem 24:2781–2795. https://doi.org/10.2174/0929867324666170303122241

    CAS  Article  PubMed  Google Scholar 

  5. 5.

    Magana-Maldonado R, Chavez-Cortez EG, Olascoaga-Arellano NK, Lopez-Mejia M, Maldonado-Leal FM, Sotelo J, Pineda B (2016) Immunological evasion in glioblastoma. Biomed Res Int 2016:7487313. https://doi.org/10.1155/2016/7487313

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  6. 6.

    Pyonteck SM, Akkari L, Schuhmacher AJ et al (2013) CSF-1R inhibition alters macrophage polarization and blocks glioma progression. Nat Med 19:1264–1272. https://doi.org/10.1038/nm.3337

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  7. 7.

    Ries CH, Cannarile MA, Hoves S et al (2014) Targeting tumor-associated macrophages with anti-CSF-1R antibody reveals a strategy for cancer therapy. Cancer Cell 25:846–859. https://doi.org/10.1016/j.ccr.2014.05.016

    CAS  Article  PubMed  Google Scholar 

  8. 8.

    Colegio OR, Chu NQ, Szabo AL et al (2014) Functional polarization of tumour-associated macrophages by tumour-derived lactic acid. Nature 513:559–563. https://doi.org/10.1038/nature13490

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  9. 9.

    Murray PJ (2017) Macrophage polarization. Annu Rev Physiol 79:541–566. https://doi.org/10.1146/annurev-physiol-022516-034339

    CAS  Article  PubMed  Google Scholar 

  10. 10.

    Sarkar S, Doring A, Zemp FJ et al (2014) Therapeutic activation of macrophages and microglia to suppress brain tumor-initiating cells. Nat Neurosci 17:46–55. https://doi.org/10.1038/nn.3597

    CAS  Article  PubMed  Google Scholar 

  11. 11.

    Gustafson MP, Lin Y, New KC, Bulur PA, O’Neill BP, Gastineau DA, Dietz AB (2010) Systemic immune suppression in glioblastoma: the interplay between CD14 + HLA-DRlo/neg monocytes, tumor factors, and dexamethasone. Neuro Oncol 12:631–644. https://doi.org/10.1093/neuonc/noq001

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  12. 12.

    Khan N, Pahari S, Vidyarthi A, Aqdas M, Agrewala JN (2016) Stimulation through CD40 and TLR-4 is an effective host directed therapy against mycobacterium tuberculosis. Front Immunol 7:386. https://doi.org/10.3389/fimmu.2016.00386

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  13. 13.

    Vidyarthi A, Khan N, Agnihotri T et al (2018) TLR-3 stimulation skews M2 macrophages to M1 through IFN-alphabeta signaling and restricts tumor progression. Front Immunol 9:1650. https://doi.org/10.3389/fimmu.2018.01650

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  14. 14.

    Khan N, Vidyarthi A, Pahari S, Negi S, Aqdas M, Nadeem S, Agnihotri T, Agrewala JN (2016) Signaling through NOD-2 and TLR-4 Bolsters the T cell priming capability of dendritic cells by inducing autophagy. Sci Rep 6:19084. https://doi.org/10.1038/srep19084

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  15. 15.

    Paul S, Calmels B, Regulier E (2002) Tumor-induced immunosuppression. Ann Biol Clin (Paris) 60:143–152

    CAS  Google Scholar 

  16. 16.

    Kostianovsky AM, Maier LM, Anderson RC, Bruce JN, Anderson DE (2008) Astrocytic regulation of human monocytic/microglial activation. J Immunol 181:5425–5432

    CAS  Article  Google Scholar 

  17. 17.

    Morantz RA, Wood GW, Foster M, Clark M, Gollahon K (1979) Macrophages in experimental and human brain tumors. Part 2: studies of the macrophage content of human brain tumors. J Neurosurg 50:305–311. https://doi.org/10.3171/jns.1979.50.3.0305

    CAS  Article  PubMed  Google Scholar 

  18. 18.

    Fritz JM, Tennis MA, Orlicky DJ et al (2014) Depletion of tumor-associated macrophages slows the growth of chemically induced mouse lung adenocarcinomas. Front Immunol 5:587. https://doi.org/10.3389/fimmu.2014.00587

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  19. 19.

    Yan D, Kowal J, Akkari L, Schuhmacher AJ, Huse JT, West BL, Joyce JA (2017) Inhibition of colony stimulating factor-1 receptor abrogates microenvironment-mediated therapeutic resistance in gliomas. Oncogene 36:6049–6058. https://doi.org/10.1038/onc.2017.261

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  20. 20.

    Martinez FO, Sica A, Mantovani A, Locati M (2008) Macrophage activation and polarization. Front Biosci 13:453–461

    CAS  Article  Google Scholar 

  21. 21.

    Schneider SW, Ludwig T, Tatenhorst L, Braune S, Oberleithner H, Senner V, Paulus W (2004) Glioblastoma cells release factors that disrupt blood-brain barrier features. Acta Neuropathol 107:272–276. https://doi.org/10.1007/s00401-003-0810-2

    Article  PubMed  Google Scholar 

  22. 22.

    Buechler C, Ritter M, Orso E, Langmann T, Klucken J, Schmitz G (2000) Regulation of scavenger receptor CD163 expression in human monocytes and macrophages by pro- and antiinflammatory stimuli. J Leukoc Biol 67:97–103

    CAS  Article  Google Scholar 

  23. 23.

    Jiang Y, Li Y, Zhu B (2015) T-cell exhaustion in the tumor microenvironment. Cell Death Dis 6:e1792. https://doi.org/10.1038/cddis.2015.162

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  24. 24.

    Huber S, Hoffmann R, Muskens F, Voehringer D (2010) Alternatively activated macrophages inhibit T-cell proliferation by Stat6-dependent expression of PD-L2. Blood 116:3311–3320. https://doi.org/10.1182/blood-2010-02-271981

    CAS  Article  PubMed  Google Scholar 

  25. 25.

    Turner NC, Reis-Filho JS (2012) Genetic heterogeneity and cancer drug resistance. Lancet Oncol 13:e178–e185. https://doi.org/10.1016/S1470-2045(11)70335-7

    Article  PubMed  Google Scholar 

  26. 26.

    Sottoriva A, Spiteri I, Piccirillo SG, Touloumis A, Collins VP, Marioni JC, Curtis C, Watts C, Tavare S (2013) Intratumor heterogeneity in human glioblastoma reflects cancer evolutionary dynamics. Proc Natl Acad Sci USA 110:4009–4014. https://doi.org/10.1073/pnas.1219747110

    Article  PubMed  Google Scholar 

  27. 27.

    Kawasaki Y, Akiyama T (2015) Tumor microenvironment: promising therapeutic target. Nihon Rinsho 73:1283–1287

    PubMed  Google Scholar 

  28. 28.

    Fahrenhold M, Rakic S, Classey J, Brayne C, Ince PG, Nicoll JAR, Boche D, Mrc C (2018) TREM2 expression in the human brain: a marker of monocyte recruitment? Brain Pathol 28:595–602. https://doi.org/10.1111/bpa.12564

    CAS  Article  PubMed  Google Scholar 

  29. 29.

    Hendrickx DAE, van Eden CG, Schuurman KG, Hamann J, Huitinga I (2017) Staining of HLA-DR, Iba1 and CD68 in human microglia reveals partially overlapping expression depending on cellular morphology and pathology. J Neuroimmunol 309:12–22. https://doi.org/10.1016/j.jneuroim.2017.04.007

    CAS  Article  PubMed  Google Scholar 

  30. 30.

    Gabrusiewicz K, Rodriguez B, Wei J et al (2016) Glioblastoma-infiltrated innate immune cells resemble M0 macrophage phenotype. JCI Insight. https://doi.org/10.1172/jci.insight.85841

    Article  PubMed  PubMed Central  Google Scholar 

  31. 31.

    Arrieta O, Montes-Servin E, Hernandez-Martinez JM, Cardona AF, Casas-Ruiz E, Crispin JC, Motola D, Flores-Estrada D, Barrera L (2017) Expression of PD-1/PD-L1 and PD-L2 in peripheral T-cells from non-small cell lung cancer patients. Oncotarget 8:101994–102005. https://doi.org/10.18632/oncotarget.22025

    Article  PubMed  PubMed Central  Google Scholar 

  32. 32.

    Jiang C, Cai X, Zhang H, Xia X, Zhang B, Xia L (2018) Activity and immune correlates of a programmed death-1 blockade antibody in the treatment of refractory solid tumors. J Cancer 9:205–212. https://doi.org/10.7150/jca.21414

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  33. 33.

    Stecher C, Battin C, Leitner J, Zettl M, Grabmeier-Pfistershammer K, Holler C, Zlabinger GJ, Steinberger P (2017) PD-1 blockade promotes emerging checkpoint inhibitors in enhancing T cell responses to allogeneic dendritic cells. Front Immunol 8:572. https://doi.org/10.3389/fimmu.2017.00572

    CAS  Article  PubMed  PubMed Central  Google Scholar 

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Acknowledgements

We thank Dr. Debajyoti Chatterjee, Gurpreet, Ishwar and Alka from Department of Histopathology, Postgraduate Institute of Medical Education and Research, Chandigarh, India for the helpful suggestions for doing immunohistochemistry.

Funding

Authors are grateful to the Council of Scientific and Industrial Research (CSIR), Department of Biotechnology (DBT), Indian Council of Medical Research (ICMR), India for financial support. Aurobind Vidyarthi and Tapan Agnihotri received fellowships from the CSIR, Nargis Khan from DBT, and Sanpreet Singh from ICMR.

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Authors

Contributions

JNA and AV conceived the project. AV, TA and NK performed experiments. AV, TA, NK, JNA, BDR, MKT and DC analyzed data. AV, JNA, NK, MKT and SS wrote the manuscript. All authors assisted in editing the manuscript and approved its final version.

Corresponding author

Correspondence to Javed N. Agrewala.

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Competing interests

All authors have declared no conflicts of interest.

Ethical approval

All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional committee. The study was approved by the Postgraduate Institute of Medical Education and Research, Chandigarh, India ‘Institutional Ethical Committee’ (Ref. No. PGI/IEC/2012/1498-99) and ‘Institutional Biosafety Committee of the Institute of Microbial Technology’ (Ref. No. 1/IEC/1/9-2014), Chandigarh, India.

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Written informed consent was obtained from all individual participants and close relative of the deceased (For cadavers) included in the study for using the specimens for research and publication.

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Vidyarthi, A., Agnihotri, T., Khan, N. et al. Predominance of M2 macrophages in gliomas leads to the suppression of local and systemic immunity. Cancer Immunol Immunother 68, 1995–2004 (2019). https://doi.org/10.1007/s00262-019-02423-8

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Keywords

  • Glioma
  • Tumor-associated macrophages
  • CD163
  • CCL3
  • PD-1