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Melanoma-conditioned medium promotes cytotoxic immune responses by murine bone marrow-derived monocytes despite their expression of ‘M2’ markers

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Abstract

Macrophages have been shown to infiltrate a wide range of malignancies and are often considered to promote tumour survival, growth and spread. However, the source and behaviour of discrete tumour-associated macrophage populations are still poorly understood. Here we show a novel method for the rational development of bone marrow-derived monocytes appropriate for the study of processes which involve the contribution of circulating inflammatory monocytes. We have shown that in response to tumour-conditioned medium, these cells upregulate CD206 and CD115, markers traditionally associated with M2-type macrophages. Treated cells show reduced capacity for cytokine secretion but significantly impact CD4+ and CD8+ T-cell proliferation and polarization. Coculture with conditioned bone marrow-derived monocytes significantly reduced CD4+ T-cell proliferation but increased CD8+ T-cell proliferation and granzyme B expression with significant induction of IFNγ secretion by both CD4+ and CD8+ T cells, indicating that these cells may have a role in promoting anti-cancer immunity.

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Abbreviations

B16CM:

B16F10-conditioned medium

BMDM:

Bone marrow-derived monocytes

iNOS:

Inducible nitric oxide synthase

mRPMI:

Macrophage RPMI-1640

NTCM:

No treatment concentrated medium

RANTES:

Regulated on activation, normal T cell expressed and secreted

RBC:

Red blood cell

TAM:

Tumour-associated macrophage

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Funding

We would like to thank Breakthrough Cancer Research who funded this research.

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Authors and Affiliations

  1. Cancer Research @ UCC, University College Cork, Fourth Floor, Western Gateway Building, Western Road, Cork, Ireland

    Liam Friel Tremble & Patrick F. Forde

  2. School of Biochemistry and Cell Biology, University College Cork, Cork, Ireland

    Anne C. Moore

Authors
  1. Liam Friel Tremble
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  2. Anne C. Moore
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  3. Patrick F. Forde
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Contributions

LFT: study design, wet lab work and paper writing. ACM and PFF: study design, evaluation of results and paper writing.

Corresponding authors

Correspondence to Liam Friel Tremble or Patrick F. Forde.

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Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval and ethical standards

All animal husbandry and experimental procedures were approved and licensed by the Animal Experimentation Ethics Committee (AEEC) in University College Cork under licence 2012-047 and performed according to the Irish Cruelty to Animals Act, 1876.

Animal source

All animals were purchased from Envigo in the UK.

Cell line authentication

The B16F10 cell line was purchased from and authenticated by the Developmental Therapeutics Program, Division of Cancer Treatment and Diagnosis Tumor Repository.

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Tremble, L.F., Moore, A.C. & Forde, P.F. Melanoma-conditioned medium promotes cytotoxic immune responses by murine bone marrow-derived monocytes despite their expression of ‘M2’ markers. Cancer Immunol Immunother 68, 1455–1465 (2019). https://doi.org/10.1007/s00262-019-02381-1

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  • DOI: https://doi.org/10.1007/s00262-019-02381-1

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