Identification and Characterization of Myeloid-Derived Suppressor Cells (MDSC) Using a Simple Approach for Conventional Flow Cytometry Analysis

Authors

  • Cátia F. Rodrigues Experimental Pathology and Therapeutics Group, Research Center of IPO-Porto (CI-IPOP), Portugal; RISE@CI-IPOP (Health Research Network), Portuguese Oncology Institute of Porto (IPO-Porto) / Porto Comprehensive Cancer Center Raquel Seruca, Porto, Portugal; Department of Immunology, Portuguese Oncology Institute of Porto (IPO-Porto), Porto, Portugal; Department of Chemistry, University of Aveiro, Aveiro, Portugal https://orcid.org/0009-0001-7005-5571
  • Catarina A. Rodrigues Experimental Pathology and Therapeutics Group, Research Center of IPO-Porto (CI-IPOP), Portugal; RISE@CI-IPOP (Health Research Network), Portuguese Oncology Institute of Porto (IPO-Porto) / Porto Comprehensive Cancer Center Raquel Seruca, Porto, Portugal; Instituto de Investigação e Inovação em Saúde (i3S), University of Porto, Porto, Portugal; Faculty of Medicine of the University of Porto (FMUP), Porto, Portugal https://orcid.org/0000-0002-1305-8055
  • Patrícia Maia Experimental Pathology and Therapeutics Group, Research Center of IPO-Porto (CI-IPOP), Portugal; RISE@CI-IPOP (Health Research Network), Portuguese Oncology Institute of Porto (IPO-Porto) / Porto Comprehensive Cancer Center Raquel Seruca, Porto, Portugal; Department of Immunology, Portuguese Oncology Institute of Porto (IPO-Porto), Porto, Portugal; Clinical Research Unit, Research Center of IPO Porto (CI-IPOP), Portuguese Oncology Institute of Porto (IPO Porto), Porto, Portugal https://orcid.org/0000-0003-1216-3280
  • M. Emília Sousa Department of Immunology, Portuguese Oncology Institute of Porto (IPO-Porto), Porto, Portugal https://orcid.org/0000-0002-4965-9530
  • Inês Godinho Department of Immunology, Portuguese Oncology Institute of Porto (IPO-Porto), Porto, Portugal https://orcid.org/0000-0001-7952-8702
  • Bruno M. Neves Department of Medical Sciences and Institute of Biomedicine (iBiMED), University of Aveiro, 3Aveiro, Portugal https://orcid.org/0000-0001-7391-3124
  • Iola F. Duarte Department of Chemistry, (CICECO & LAQV/REQUIMTE), University of Aveiro, Aveiro, Portugal https://orcid.org/0000-0003-4289-9256
  • Lúcio Lara Santos Experimental Pathology and Therapeutics Group, Research Center of IPO-Porto (CI-IPOP), Portugal; RISE@CI-IPOP (Health Research Network), Portuguese Oncology Institute of Porto (IPO-Porto) / Porto Comprehensive Cancer Center Raquel Seruca, Porto, Portugal; Health School of University Fernando Pessoa, Porto, Portugal; Department of Surgical Oncology, Portuguese Oncology Institute of Porto (IPO Porto), Porto, Portugal https://orcid.org/0000-0002-0521-5655
  • Carlos Palmeira Experimental Pathology and Therapeutics Group, Research Center of IPO-Porto (CI-IPOP), Portugal; RISE@CI-IPOP (Health Research Network), Portuguese Oncology Institute of Porto (IPO-Porto) / Porto Comprehensive Cancer Center Raquel Seruca, Porto, Portugal; Department of Immunology, Portuguese Oncology Institute of Porto (IPO-Porto), Porto, Portugal; Health School of University Fernando Pessoa, Porto, Portugal https://orcid.org/0000-0002-4833-2202
  • Gabriela Martins Department of Immunology, Portuguese Oncology Institute of Porto (IPO-Porto), Porto, Portugal https://orcid.org/0000-0002-1367-8852

DOI:

https://doi.org/10.34635/rpc.1099

Keywords:

Biomarkers, Tumor, Flow Cytometry, Myeloid-Derived Suppressor Cells, Neoplasms

Abstract

Introduction: Myeloid-derived suppressor cells (MDSCs) are a heterogeneous population of immature myeloid cells that accumulate under pathological conditions, such as cancer, and suppress immune responses through various mechanisms. Distinguishing MDSC populations remains challenging due to their shared surface markers with neutrophils and monocytes, complicating accurate identification and quantification. This study aimed to improve MDSC identification and quantification using conventional flow cytometry panels and assess their functional activity for integration into the cancer immunogram of cancer patients at the Immunology Department of IPO-Porto.

Methods: Identification and quantification of circulating MDSCs were performed by flow cytometry using an 8-color multiparametric panel.

Results: Cancer patients showed significantly higher levels of PMN-MDSCs (12-fold) and M-MDSCs (1.1-fold) than healthy donors. MDSC function was evaluated by qPCR after cell sorting (FACS), revealing increased transcriptional levels of NOS2 and TGFB1, which are associated with immunosuppressive activity. Moreover, reduced zeta chain (CD247) expression in T lymphocytes and NK cells was observed, with lower mean fluorescence intensity (MFI) ratios in cancer patients, indicating impaired immune signaling.

Conclusion: This study confirmed that circulating MDSC levels are elevated in cancer patients, reinforcing their relevance in the cancer immunogram. It also identified potentially useful phenotypic and functional MDSC markers that require validation in larger sample sets. The findings contribute to refining flow cytometry analysis panels, enabling more accurate and standardized identification of MDSC populations using conventional platforms.

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References

Brown JS, Amend SR, Austin RH, Gatenby RA, Hammarlund EU, Pienta KJ. Updating the Definition of Cancer. Mol Cancer Res. 2023;21:142-47.

Gupta I, Hussein O, Sastry KS, Bougarn S, Gopinath N, Chin-Smith E, et al. Deciphering the complexities of cancer cell immune evasion: Mechanisms and therapeutic implications. Adv Cancer Biology - Metastasis. 2023;8:100107. doi:10.1016/j.addanc.2023.100107

Blank CU, Haanen JB, Ribas A, Schumacher TN. CANCER IMMUNOLOGIC: The "cancer immunogram". Science. 2016;352: 658-60.

Mellman I, Chen DS, Powles T, Turley SJ. The cancer-immunity cycle: Indication, genotype, and immunotype. Immunity. 2023;56: 2188-205.

Yang K, Lu R, Mei J, Cao K, Zeng T, Hua Y, et al. The war between the immune system and the tumor – using immune biomarkers as tracers. Biomark Res. 2024;12:51. doi:10.1186/s40364-024-00599-5.

Kim SJ, Cassella CR, Byrne KT. Tumor burden and immunotherapy: Impact on immune infiltration and therapeutic outcomes. Front Immunol. 2021;11:629722. doi:10.3389/fimmun.2020.629722.

He S, Zheng L, Qi C. Myeloid-derived suppressor cells (MDSCs) in the tumor microenvironment and their targeting in cancer therapy. Mol Cancer. 2025;24:5. doi:10.1186/s12943-024-02208-3.

Liu T, Rosiek A, Gonzalez De Los Santos F, Phan SH. Chapter 1 – Detection of myeloid-derived suppressor cells by flow cytometry. In: Jiménez-Cortegana C, Galluzzi L, editors. Methods in Cell Biology. London: Academic Press; 2024. p.1-15.

Li K, Shi H, Zhang B, Ou X, Ma Q, Chen Y, et al. Myeloid-derived suppressor cells as immunosuppressive regulators and therapeutic targets in cancer. Signal Transduct Target Ther. 2021;6:362. doi:10.1038/s41592-021-00670-9.

Glover A, Zhang Z, Shannon-Lowe C. Deciphering the roles of myeloid derived suppressor cells in viral oncogenesis. Front Immunol. 2023;14:1161848.

Wang S, Zhao X, Wu S, Cui D, Xu Z. Myeloid-derived suppressor cells: key immunosuppressive regulators and therapeutic targets in hematological malignancies. Biomarker Res. 2023;11:34.

Raskov H, Orhan A, Gaggar S, Gögenur I. Neutrophils and polymorphonuclear myeloid-derived suppressor cells: an emerging battleground in cancer therapy. Oncogenesis. 2022;11:22.

Groth C, Weber R, Lasser S, Özbay FG, Kurzay A, Petrova V, et al. Tumor promoting capacity of polymorphonuclear myeloid-derived suppressor cells and their neutralization. Int J Cancer. 2021;149:1628-38. doi:10.1002/jic.33731.

Davidov V, Jensen G, Mai S, Chen S-H, Pan PY. Analyzing one cell at a TIME: Analysis of myeloid cell contributions in the tumor immune microenvironment. Front Immunol.2021;11:645213. doi:10.3389/fimmun.2020.645213.

De Cicco P, Ercolano G, lanaro A. The new era of cancer immunotherapy: targeting myeloid-derived suppressor cells to overcome immune evasion. Front Immunol. 2020;11:1680. doi:10.3389/fimmun.2020.01680.

Shi H, Li K, Ni Y, Liang X, Zhao X. Myeloid-derived suppressor cells: Implications in the resistance of malignant tumors to T cell-based immunotherapy. Front Cell Dev Biol. 2021;9:707198. doi:10.3389/fcell.2021.707198.

Gabrilovich DI. Myeloid-Derived Suppressor Cells. Cancer Immunol Res. 2017;5:3-8.

Ge Y, Cheng D, Jia Q, Xiong H, Zhang J. Mechanisms underlying the role of myeloid-derived suppressor cells in clinical diseases: good or bad. Immune Netw. 2021;21:e21. doi:10.4110/in.2021.21.e21.

Bronte V, Brandau S, Chen SH, Colombo MP, Frey AB, Greten TF, et al. Recommendations for myeloid-derived suppressor cell nomenclature and characterization standards. Nat Commun. 2016;7:2150. doi:10.1038/ncomms12150.

Ni M, Cui J, Yang X, Ding Y, Zhao P, Hu T, et al. Dual roles of CD11bCD33HLA-DR/devCD14* myeloid-derived suppressor cells with a granulocytic morphology following allogeneic hematopoietic stem cell transplantation: from inflammation promoters to immune suppressors within 90 days. Front Immunol. 2024;15:1403272. doi:10.3389/fimmu.2024.1403272.

Coudereau R, Waeckel L, Cour M, Rimmel E T, Pescarmona R, Fabri A, et al. Emergence of immunosuppressive LOX-1+ PMN-MDSC in septic shock and severe COVID-19 patients with acute respiratory distress syndrome. J Leukoc Biol. 2022;111489-96. doi:10.1002/JLB.4COVBCR0321-129R.

Sanchez-Pino MD, Dean MJ, Ochoa AC. Myeloid-derived suppressor cells (MDSC): When good intentions go awry. Cell Immunol. 2021;362:104302.

Tian X, Wang T, Zheng Q, Tao Y, Dai L, Shen H. Circulating CD15(t) LOX-1(t)- PMN-MDSCs are a potential biomarker for the early diagnosis of non-small-cell lung cancer. Int J Clin Pract. 2021;75:e14317.

Lasser SA, Ozbay Kurt FG, Arkhypov I, Utikal J, Umansky V. Myeloid-derived suppressor cells in cancer and cancer therapy. Nat Rev Clin Oncol. 2024;21:147-64. doi:10.1038/s41571-023-00846-y.

Bruderek K, Schirrmann R, Brandau S. Immunophenotyping of Circulating Myeloid-Derived Suppressor Cells (MDSC) in the Peripheral Blood of Cancer Patients. Methods Mol Biol. 2021;223:61-7. doi:10.1007/978-1-0716-1060-2_1.

Condamine T, Dominguez GA, Youm JJ, Kossenkov AV, Mony S, Alicea-Torres K, et al. Lectin-type oxidized LDL receptor-1 distinguishes population of human polymorphonuclear myeloid-derived suppressor cells in cancer patients. Sci Immunol. 2016;1aaf8943. doi:10.1126/sciimmunol.aaf8943.

Cassetta L, Baekkevold ES, Brandau S, Bujko A, Cassatella MA, Dorhoi A, et al. Deciphering myeloid-derived suppressor cells: isolation and markers in humans, mice and non-human primates. Cancer Immunol Immunother. 2019;68:687-97. doi:10.1007/s00262-019-02302-2.

Cassetta L, Bruderek K, Skrzeczynska-Moncznik J, Osiecka O, Hu X, Rundgren IM, et al. Differential expansion of circulating human MDSC subsets in patients with cancer, infection and inflammation. J Immunother Cancer. 2020;8:e001223. doi:10.1136/jitc.2020-001223.

Ostrand-Rosenberg S, Lamb TJ, Pawelec G. Here, There, and Everywhere: Myeloid-Derived Suppressor Cells in Immunology. J Immunol. 2023;210:1183-97.

Ren R, Xiong C, Ma R, Wang Y, Yue T, Yu J, et al. The recent progress of myeloid-derived suppressor cell and its targeted therapies in cancers. MedComm. 2023;4:e323. doi:10.1002/mco2.323.

Zarobkiewicz M, Kowalska W, Chocholska S, Tomczak W, Szymańska A, Morawska I, et al. High M-MDSC Percentage as a Negative Prognostic Factor in Chronic Lymphocytic Leukaemia. Cancers. 2020;12:2614. doi:10.3390/cancers12092614.

Ren W, Zhang X, Li W, Feng Q, Feng H, Tong Y, et al. Circulating and tumor-infiltrating arginate 1-expressing cells in gastric adenocarcinoma patients were mainly immature and monocytic Myeloid-derived suppressor cells. Sci Rep. 2020;10(1):8056. doi:10.1038/s41598-020-64841-4.

Ye W, Zhou Y, Xu B, et al. CD247 expression is associated with differentiation and classification in ovarian cancer. Medicine. 2019;98:e18407.

Reuven O, Mikula I, Jr., Ashkenazi-Preiser H, Twaik N, Ben-Meir K, Meirow Y, et al. Functional assays evaluating immunosuppression mediated by myeloid-derived suppressor cells. Curr Protoc. 2022;2:e557. doi:10.1002/cpxl.557.

Joshi S, Sharabi A. Targeting myeloid-derived suppressor cells to enhance natural killer cell-based immunotherapy. Pharmacol Ther. 2022;235:108114. doi:10.1016/j.pharmthera.2022.108114.

Hwang JR, Byeon Y, Kim D, Park SG. Recent insights of T cell receptor-mediated signaling pathways for T cell activation and development. Exp Mol Med. 2020;52:750-61. doi:10.1038/s12276-020-0435-8.

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Published

2025-06-09

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Original Papers