Recommendations for myeloid-derived suppressor cell nomenclature and characterization standards

Vincenzo Bronte, Sven Brandau, Shu-Hsia Chen, Mario P Colombo, Alan B Frey, Tim F Greten, Susanna Mandruzzato, Peter J Murray, Augusto Ochoa, Suzanne Ostrand-Rosenberg, Paulo C Rodriguez, Antonio Sica, Viktor Umansky, Robert H Vonderheide, Dmitry I Gabrilovich, Vincenzo Bronte, Sven Brandau, Shu-Hsia Chen, Mario P Colombo, Alan B Frey, Tim F Greten, Susanna Mandruzzato, Peter J Murray, Augusto Ochoa, Suzanne Ostrand-Rosenberg, Paulo C Rodriguez, Antonio Sica, Viktor Umansky, Robert H Vonderheide, Dmitry I Gabrilovich

Abstract

Myeloid-derived suppressor cells (MDSCs) have emerged as major regulators of immune responses in cancer and other pathological conditions. In recent years, ample evidence supports key contributions of MDSC to tumour progression through both immune-mediated mechanisms and those not directly associated with immune suppression. MDSC are the subject of intensive research with >500 papers published in 2015 alone. However, the phenotypic, morphological and functional heterogeneity of these cells generates confusion in investigation and analysis of their roles in inflammatory responses. The purpose of this communication is to suggest characterization standards in the burgeoning field of MDSC research.

Figures

Figure 1. Gating strategy for the identification…
Figure 1. Gating strategy for the identification of mouse MDSC subsets.
Gating strategy used to define MDSC subpopulations in BM, blood and spleen of C57Bl/6 tumour-free or MCA203 tumour-bearing mice. After exclusion of doublets (not shown), live CD11b+ cells were gated and the proportion of Ly6C and Ly6G cells was evaluated.
Figure 2. Gating strategy for the identification…
Figure 2. Gating strategy for the identification of MDSC subsets in the peripheral blood of healthy donors and melanoma patients.
Doublets were excluded and live PBMC were gated (not shown). (a) CD14+HLA-DR−/lo M-MDSC. Monocytes were gated on the basis of FSC and SSC parameters and HLA-DR downregulation was defined by FMO control. (b) Lin−HLA-DR−CD33+ eMDSC. (c) CD14−CD15+CD11b+ PMN-MDSC.
Figure 3. Overview of MDSC involvement in…
Figure 3. Overview of MDSC involvement in myeloid cell differentiation in cancer.
In cancer and chronic inflammation, the bone marrow and spleen increase the output of mature and immature myeloid cells that comprise a spectrum between monocytes and neutrophils. In mice, MDSC toward the monocytic end of the spectrum (M-MDSC) are CD11b+Ly6C+Ly6G−, while towards the neutrophil end of the spectrum (PMN-MDSC) are CD11b+LyG+Ly6C−. Within solid tumours M-MDSC develop through intermediate steps towards macrophages where Ly6C is progressively downregulated and MHCII, F4/80 and CX3CR1 are upregulated. Under chronic inflammation, monocytic lineages show an increasing requirement for anti-apoptotic survival pathways (mediated primarily by GM-CSF signalling) to block the intrinsic mitochondrial death pathway. A similar scheme is likely to occur in humans; however, the cell markers are different.
Figure 4. Algorithm for identification of cells…
Figure 4. Algorithm for identification of cells as MDSC.
Step-by-step approach to identify cells as MDSC for reporting. It is important, wherever possible, to use cells with the same phenotype from control mice or healthy donors as controls.

References

    1. Brusa D. et al. Circulating immunosuppressive cells of prostate cancer patients before and after radical prostatectomy: profile comparison. Int. J. Urol. 20, 971–978 (2013).
    1. Bronte V. et al. Apoptotic death of CD8+ T lymphocytes after immunization: induction of a suppressive population of Mac-1+/Gr-1+ cells. J. Immunol. 161, 5313–5320 (1998).
    1. Gabrilovich D. et al. Vascular endothelial growth factor inhibits the development of dendritic cells and dramatically affects the differentiation of multiple hematopoietic lineages in vivo. Blood 92, 4150–4166 (1998).
    1. Pekarek L. A., Starr B. A., Toledano A. Y. & Schreiber H. Inhibition of tumor growth by elimination of granulocytes. J. Exp. Med. 181, 435–440 (1995).
    1. Talmadge J. E. & Gabrilovich D. I. History of myeloid-derived suppressor cells. Nat. Rev. Cancer 13, 739–752 (2013).
    1. Gabrilovich D. I. et al. The terminology issue for myeloid-derived suppressor cells. Cancer Res. 67, 425 (2007).
    1. Movahedi K. et al. Identification of discrete tumor-induced myeloid-derived suppressor cell subpopulations with distinct T cell-suppressive activity. Blood 111, 4233–4244 (2008).
    1. Youn J. I., Nagaraj S., Collazo M. & Gabrilovich D. I. Subsets of myeloid-derived suppressor cells in tumor-bearing mice. J. Immunol. 181, 5791–5802 (2008).
    1. Youn J. I., Collazo M., Shalova I. N., Biswas S. K. & Gabrilovich D. I. Characterization of the nature of granulocytic myeloid-derived suppressor cells in tumor-bearing mice. J. Leukoc. Biol. 91, 167–181 (2012).
    1. Rodriguez P. C. et al. Arginase I-producing myeloid-derived suppressor cells in renal cell carcinoma are a subpopulation of activated granulocytes. Cancer Res. 69, 1553–1560 (2009).
    1. Solito S. et al. Myeloid-derived suppressor cell heterogeneity in human cancers. Ann. NY Acad. Sci. 1319, 47–65 (2014).
    1. Marvel D. & Gabrilovich D. I. Myeloid-derived suppressor cells in the tumor microenvironment: expect the unexpected. J. Clin. Invest. 125, 3356–3364 (2015).
    1. Gabrilovich D. I., Ostrand-Rosenberg S. & Bronte V. Coordinated regulation of myeloid cells by tumours. Nat. Rev. Immunol. 12, 253–268 (2012).
    1. Peranzoni E. et al. Myeloid-derived suppressor cell heterogeneity and subset definition. Curr. Opin. Immunol. 22, 238–244 (2010).
    1. Dumitru C. A., Moses K., Trellakis S., Lang S. & Brandau S. Neutrophils and granulocytic myeloid-derived suppressor cells: immunophenotyping, cell biology and clinical relevance in human oncology. Cancer Immunol. Immunother. 61, 1155–1167 (2012).
    1. Mandruzzato S. et al. Toward harmonized phenotyping of human myeloid-derived suppressor cells by flow cytometry: results from an interim study. Cancer Immunol. Immunother. 65, 161–169 (2016).
    1. Liu T., Daniels C. K. & Cao S. Comprehensive review on the HSC70 functions, interactions with related molecules and involvement in clinical diseases and therapeutic potential. Pharmacol. Ther. 136, 354–374 (2012).
    1. Hirai H. et al. C/EBP[beta] is required for 'emergency' granulopoiesis. Nat. Immunol. 7, 732–739 (2006).
    1. Trellakis S. et al. Granulocytic myeloid-derived suppressor cells are cryosensitive and their frequency does not correlate with serum concentrations of colony-stimulating factors in head and neck cancer. Innate. Immun. 19, 328–336 (2013).
    1. Viale A. et al. Oncogene ablation-resistant pancreatic cancer cells depend on mitochondrial function. Nature 514, 628–632 (2014).
    1. Dolcetti L. et al. Hierarchy of immunosuppressive strength among myeloid-derived suppressor cell subsets is determined by GM-CSF. Eur. J. Immunol. 40, 22–35 (2010).
    1. Elkabets M. et al. IL-1beta regulates a novel myeloid-derived suppressor cell subset that impairs NK cell development and function. Eur. J. Immunol. 40, 3347–3357 (2010).
    1. Hongo D., Tang X., Baker J., Engleman E. G. & Strober S. Requirement for interactions of natural killer T cells and myeloid-derived suppressor cells for transplantation tolerance. Am. J. Transplant. 14, 2467–2477 (2014).
    1. Li Y. et al. Myeloid-derived suppressor cells as a potential therapy for experimental autoimmune myasthenia gravis. J. Immunol. 193, 2127–2134 (2014).
    1. Koehn B. H. et al. GVHD-associated, inflammasome-mediated loss of function in adoptively transferred myeloid-derived suppressor cells. Blood 126, 1621–1628 (2015).
    1. Marigo I. et al. Tumor-induced tolerance and immune suppression depend on the C/EBPbeta transcription factor. Immunity 32, 790–802 (2010).
    1. Nagaraj S. et al. Anti-inflammatory triterpenoid blocks immune suppressive function of MDSCs and improves immune response in cancer. Clin. Cancer Res. 16, 1812–1823 (2010).
    1. Youn J. I. et al. Epigenetic silencing of retinoblastoma gene regulates pathologic differentiation of myeloid cells in cancer. Nat. Immunol. 14, 211–220 (2013).
    1. Svensson R. U. et al. Slow disease progression in a C57BL/6 pten-deficient mouse model of prostate cancer. Am. J. Pathol. 179, 502–512 (2011).
    1. Lavin Y. et al. Tissue-resident macrophage enhancer landscapes are shaped by the local microenvironment. Cell 159, 1312–1326 (2014).
    1. Sonda N. et al. miR-142-3p prevents macrophage differentiation during cancer-induced myelopoiesis. Immunity 38, 1236–1249 (2013).
    1. Rigamonti N. et al. Modulators of arginine metabolism do not impact on peripheral T-cell tolerance and disease progression in a model of spontaneous prostate cancer. Clin. Cancer. Res. 17, 1012–1023 (2011).
    1. Spranger S. et al. Up-regulation of PD-L1, IDO, and T(regs) in the melanoma tumor microenvironment is driven by CD8(+) T cells. Sci. Transl. Med. 5, 200ra116 (2013).
    1. DeNardo D. G. et al. Leukocyte complexity predicts breast cancer survival and functionally regulates response to chemotherapy. Cancer Discov. 1, 54–67 (2011).
    1. Gentles A. J. et al. The prognostic landscape of genes and infiltrating immune cells across human cancers. Nat. Med. 21, 938–945 (2015).
    1. Ugel S., De Sanctis F., Mandruzzato S. & Bronte V. Tumor-induced myeloid deviation: when myeloid-derived suppressor cells meet tumor-associated macrophages. J. Clin. Invest. 125, 3365–3376 (2015).
    1. Noy R. & Pollard J. W. Tumor-associated macrophages: from mechanisms to therapy. Immunity 41, 49–61 (2014).
    1. Fridlender Z. G. et al. Polarization of tumor-associated neutrophil phenotype by TGF-beta: "N1" versus "N2" TAN. Cancer Cell 16, 183–194 (2009).
    1. Murray P. J. et al. Macrophage activation and polarization: nomenclature and experimental guidelines. Immunity 41, 14–20 (2014).
    1. Sica A. & Mantovani A. Macrophage plasticity and polarization: in vivo veritas. J. Clin. Invest. 122, 787–795 (2012).
    1. Kratochvill F. et al. TNF counterbalances the emergence of M2 tumor macrophages. Cell Rep. 12, 1902–1914 (2015).
    1. Movahedi K. et al. Different tumor microenvironments contain functionally distinct subsets of macrophages derived from Ly6C(high) monocytes. Cancer Res. 70, 5728–5739 (2010).
    1. Haverkamp J. M. et al. Myeloid-derived suppressor activity is mediated by monocytic lineages maintained by continuous inhibition of extrinsic and intrinsic death pathways. Immunity 41, 947–959 (2014).
    1. Strauss L. et al. RORC1 regulates tumor-promoting "Emergency" granulo-monocytopoiesis. Cancer Cell 28, 253–269 (2015).
    1. Kumar V., Patel S., Tcyganov E. & Gabrilovich D. I. The nature of myeloid-derived suppressor cells in the tumor microenvironment. Trends Immunol. 37, 208–220 (2016).
    1. Thevenot P. T. et al. The stress-response sensor chop regulates the function and accumulation of myeloid-derived suppressor cells in tumors. Immunity 41, 389–401 (2014).
    1. Condamine T. et al. ER stress regulates myeloid-derived suppressor cell fate through TRAIL-R-mediated apoptosis. J. Clin. Invest. 124, 2626–2639 (2014).
    1. Fridlender Z. G. et al. Transcriptomic analysis comparing tumor-associated neutrophils with granulocytic myeloid-derived suppressor cells and normal neutrophils. PLoS ONE 7, e31524 (2012).
    1. Eruslanov E. B. et al. Tumor-associated neutrophils stimulate T cell responses in early-stage human lung cancer. J. Clin. Invest. 124, 5466–5480 (2014).
    1. Mishalian I., Granot Z. & Fridlender Z. G. The diversity of circulating neutrophils in cancer. Immunobiology S0171–2985, 30017–1 (2016).
    1. Cimen Bozkus C., Elzey B. D., Crist S. A., Ellies L. G. & Ratliff T. L. Expression of cationic amino acid transporter 2 is required for myeloid-derived suppressor cell-mediated control of T cell immunity. J. Immunol. 195, 5237–5250 (2015).
    1. Mairhofer D. G. et al. Impaired gp100-Specific CD8(+) T-cell responses in the presence of myeloid-derived suppressor cells in a spontaneous mouse melanoma model. J. Invest. Dermatol. 135, 2785–2793 (2015).
    1. Raber P. L. et al. Subpopulations of myeloid-derived suppressor cells impair T cell responses through independent nitric oxide-related pathways. Int. J. Cancer 134, 2853–2864 (2014).
    1. Mantovani A. & Sica A. Macrophages, innate immunity and cancer: balance, tolerance, and diversity. Curr. Opin. Immunol. 22, 231–237 (2010).
    1. Ortiz M. L., Lu L., Ramachandran I. & Gabrilovich D. I. Myeloid-derived suppressor cells in the development of lung cancer. Cancer Immunol. Res. 2, 50–58 (2014).
    1. Waight J. D. et al. Myeloid-derived suppressor cell development is regulated by a STAT/IRF-8 axis. J. Clin. Invest. 123, 4464–4478 (2013).
    1. Chalmin F. et al. Membrane-associated Hsp72 from tumor-derived exosomes mediates STAT3-dependent immunosuppressive function of mouse and human myeloid-derived suppressor cells. J. Clin. Invest. 120, 457–471 (2010).
    1. Nefedova Y. et al. Hyperactivation of STAT3 is involved in abnormal differentiation of dendritic cells in cancer. J. Immunol. 172, 464–474 (2004).
    1. Corzo C. A. et al. Mechanism regulating reactive oxygen species in tumor-induced myeloid-derived suppressor cells. J. Immunol. 182, 5693–5701 (2009).
    1. Sinha P. et al. Proinflammatory s100 proteins regulate the accumulation of myeloid-derived suppressor cells. J. Immunol. 181, 4666–4675 (2008).
    1. Cheng P. et al. Inhibition of dendritic cell differentiation and accumulation of myeloid-derived suppressor cells in cancer is regulated by S100A9 protein. J. Exp. Med. 205, 2235–2249 (2008).
    1. Finke J. et al. MDSC as a mechanism of tumor escape from sunitinib mediated anti-angiogenic therapy. Int. Immunopharmacol. 11, 856–861 (2011).
    1. Gallina G. et al. Tumors induce a subset of inflammatory monocytes with immunosuppressive activity on CD8+ T cells. J. Clin. Invest. 116, 2777–2790 (2006).
    1. Bronte V. et al. IL-4-induced arginase 1 suppresses alloreactive T cells in tumor-bearing mice. J. Immunol. 170, 270–278 (2003).
    1. Molon B. et al. Chemokine nitration prevents intratumoral infiltration of antigen-specific T cells. J. Exp. Med. 208, 1949–1962 (2011).
    1. Umemura N. et al. Tumor-infiltrating myeloid-derived suppressor cells are pleiotropic-inflamed monocytes/macrophages that bear M1- and M2-type characteristics. J. Leukoc. Biol. 83, 1136–1144 (2008).
    1. Nagaraj S., Schrum A. G., Cho H. I., Celis E. & Gabrilovich D. I. Mechanism of T cell tolerance induced by myeloid-derived suppressor cells. J. Immunol. 184, 3106–3116 (2010).
    1. Lu T. et al. Tumor-infiltrating myeloid cells induce tumor cell resistance to cytotoxic T cells in mice. J. Clin. Invest. 121, 4015–4029 (2011).
    1. Shojaei F. et al. Tumor refractoriness to anti-VEGF treatment is mediated by CD11b(+)Gr1(+) myeloid cells. Nat. Biotechnol. 25, 911–920 (2007).
    1. Yang L. et al. Expansion of myeloid immune suppressor Gr+CD11b+ cells in tumor-bearing host directly promotes tumor angiogenesis. Cancer Cell 6, 409–421 (2004).
    1. Sinha P., Clements V. K., Fulton A. M. & Ostrand-Rosenberg S. Prostaglandin E2 promotes tumor progression by inducing myeloid-derived suppressor cells. Cancer. Res. 67, 4507–4513 (2007).
    1. Mao Y. et al. Inhibition of tumor-derived prostaglandin-e2 blocks the induction of myeloid-derived suppressor cells and recovers natural killer cell activity. Clin. Cancer. Res. 20, 4096–4106 (2014).
    1. Noman M. Z. et al. PD-L1 is a novel direct target of HIF-1alpha, and its blockade under hypoxia enhanced MDSC-mediated T cell activation. J. Exp. Med. 211, 781–790 (2014).
    1. Beury D. W. et al. Cross-talk among myeloid-derived suppressor cells, macrophages, and tumor cells impacts the inflammatory milieu of solid tumors. J. Leukoc. Biol. 96, 1109–1118 (2014).
    1. Sinha P., Clements V. K., Bunt S. K., Albelda S. M. & Ostrand-Rosenberg S. Cross-talk between myeloid-derived suppressor cells and macrophages subverts tumor immunity toward a type 2 response. J. Immunol. 179, 977–983 (2007).
    1. Sumida K. et al. Anti-IL-6 receptor mAb eliminates myeloid-derived suppressor cells and inhibits tumor growth by enhancing T-cell responses. Eur. J. Immunol. 42, 2060–2072 (2012).
    1. Roth F. et al. Aptamer-mediated blockade of IL4Ralpha triggers apoptosis of MDSCs and limits tumor progression. Cancer Res. 72, 1373–1383 (2012).
    1. Serafini P. et al. High-dose GM-CSF-producing vaccines impair the immune response through the recruitment of myeloid suppressor cells. Cancer Res. 64, 6337–6343 (2004).
    1. Abe F. et al. Myeloid-derived suppressor cells in mammary tumor progression in FVB Neu transgenic mice. Cancer Immunol. Immunother. 59, 47–62 (2010).
    1. Kowanetz M. et al. Granulocyte-colony stimulating factor promotes lung metastasis through mobilization of Ly6G+Ly6C+ granulocytes. Proc. Natl Acad. Sci. USA 107, 21248–21255 (2010).
    1. Highfill S. L. et al. Bone marrow myeloid-derived suppressor cells (MDSCs) inhibit graft-versus-host disease (GVHD) via an arginase-1-dependent mechanism that is up-regulated by interleukin-13. Blood 116, 5738–5747 (2010).

Source: PubMed

Sponsorer og samarbejdspartnere

Medicinske tilstande

Narkotikainterventioner

3
Abonner