title: MIF inhibition reverts the gene expression profile of human melanoma cell line-induced MDSCs to normal monocytes.
abstract: Myeloid-derived suppressor cells (MDSCs) are potently immunosuppressive innate immune cells that accumulate in advanced cancer patients and actively inhibit anti-tumor T lymphocyte responses [1]. Increased numbers of circulating MDSCs directly correlate with melanoma patient morbidity and reduced anti-tumor immune responses [2], [3]. Previous studies have revealed that monocyte-derived macrophage migration inhibitory factor (MIF) is necessary for the immune suppressive function of MDSCs in mouse models of melanoma [4], [5]. To investigate whether MIF participates in human melanoma-induced MDSC differentiation and/or suppressive function, we have established an in vitro MDSC induction model using primary, normal human monocytes co-cultured with human melanoma cell lines in the presence or absence of the MIF antagonist-4-IPP [4], [6], [7], [8], [9]. To identify potential mechanistic effectors, we have performed transcriptome analyses on cultured monocytes and on melanoma-induced MDSCs obtained from either untreated or 4-IPP-treated A375:monocyte co-cultures. Here, we present a detailed protocol, which can facilitate easy reproduction of the microarray results (NCBI GEO accession number GSE73333) published by Yaddanapudi et al. (2015) in Cancer Immunology Research [10].
authors.affiliation: Molecular Targets Program, JG Brown Cancer Center, University of Louisville, Louisville, KY 40202, USA.
authors.forename: Sabine
authors.initials: S
authors.lastname: Waigel
authors.affiliation: Molecular Targets Program, JG Brown Cancer Center, University of Louisville, Louisville, KY 40202, USA.
authors.forename: Beatriz E
authors.initials: BE
authors.lastname: Rendon
authors.affiliation: Molecular Targets Program, JG Brown Cancer Center, University of Louisville, Louisville, KY 40202, USA.
authors.forename: Gwyneth
authors.initials: G
authors.lastname: Lamont
authors.affiliation: Molecular Targets Program, JG Brown Cancer Center, University of Louisville, Louisville, KY 40202, USA.
authors.forename: Jamaal
authors.initials: J
authors.lastname: Richie
authors.affiliation: Molecular Targets Program, JG Brown Cancer Center, University of Louisville, Louisville, KY 40202, USA; Department of Microbiology and Immunology, University of Louisville, USA; Department of Medicine, University of Louisville, USA.
authors.forename: Robert A
authors.initials: RA
authors.lastname: Mitchell
authors.affiliation: Molecular Targets Program, JG Brown Cancer Center, University of Louisville, Louisville, KY 40202, USA; Department of Microbiology and Immunology, University of Louisville, USA; Department of Medicine, University of Louisville, USA.
authors.forename: Kavitha
authors.initials: K
authors.lastname: Yaddanapudi
keywords: Immunesuppression; MDSC; MIF; Melanoma; Trancriptome analysis