Abstract
Myeloid derived suppressor cells (MDSCs) from tumor-bearing mice are important negative regulators of anti-cancer immune responses, but the role for immature myeloid cells (IMCs) in non-tumor-bearing mice in the regulation of immune responses are poorly described. We studied the immune-suppressive activity of IMCs from the bone marrow (BM) of C57Bl/6 mice and the mechanism(s) by which they inhibit T-cell activation and proliferation. IMCs, isolated from BM by high-speed FACS, inhibited mitogen-induced proliferation of CD4(+) and CD8(+) T-cells in vitro. Cell-to-cell contact of T-cells with viable IMCs was required for suppression. Neither neutralizing antibodies to TGFβ1, nor genetic disruption of indolamine 2,3-dioxygenase, abrogated IMC-mediated suppressive activity. In contrast, suppression of T-cell proliferation was absent in cultures containing IMCs from interferon-γ (IFN-γ) receptor KO mice or T-cells from IFN-γ KO mice (on the C57Bl/6 background). The addition of NO inhibitors to co-cultures of T-cells and IMC significantly reduced the suppressive activity of IMCs. IFN-γ signaling between T-cells and IMCs induced paracrine Nitric Oxide (NO) release in culture, and the degree of inhibition of T-cell proliferation was proportional to NO levels. The suppressive activity of IMCs from the bone marrow of tumor-free mice was comparable with MDSCs from BALB/c bearing mice 4T1 mammary tumors. These results indicate that IMCs have a role in regulating T-cell activation and proliferation in the BM microenvironment.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Animals
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Antibodies, Neutralizing / pharmacology
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Bone Marrow Cells / cytology*
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Bone Marrow Cells / immunology
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Bone Marrow Cells / metabolism
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CD4-Positive T-Lymphocytes / cytology
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CD4-Positive T-Lymphocytes / immunology
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CD4-Positive T-Lymphocytes / metabolism
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CD8-Positive T-Lymphocytes / cytology
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CD8-Positive T-Lymphocytes / immunology
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CD8-Positive T-Lymphocytes / metabolism
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Cell Communication / drug effects
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Cell Differentiation / drug effects
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Cell Proliferation / drug effects
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Coculture Techniques
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Enzyme Inhibitors / pharmacology
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Female
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Gene Expression
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Indoleamine-Pyrrole 2,3,-Dioxygenase / deficiency
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Indoleamine-Pyrrole 2,3,-Dioxygenase / genetics
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Interferon-gamma / deficiency
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Interferon-gamma / genetics
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Mammary Glands, Animal / immunology
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Mammary Glands, Animal / metabolism
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Mammary Glands, Animal / pathology*
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Mammary Neoplasms, Experimental / immunology
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Mammary Neoplasms, Experimental / metabolism
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Mammary Neoplasms, Experimental / pathology*
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Mice
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Mice, Inbred C57BL
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Mice, Knockout
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Myeloid Cells / cytology*
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Myeloid Cells / immunology
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Myeloid Cells / metabolism
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Nitric Oxide / antagonists & inhibitors
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Nitric Oxide / immunology
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Nitric Oxide / metabolism*
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Signal Transduction
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Transforming Growth Factor beta1 / antagonists & inhibitors
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Transforming Growth Factor beta1 / biosynthesis
Substances
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Antibodies, Neutralizing
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Enzyme Inhibitors
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Indoleamine-Pyrrole 2,3,-Dioxygenase
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Transforming Growth Factor beta1
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Nitric Oxide
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Interferon-gamma
Grants and funding
The authors have no support or funding to report.