title: An optimized protocol for human M2 macrophages using M-CSF and IL-4/IL-10/TGF-β yields a dominant immunosuppressive phenotype.
abstract: Monocytes are highly abundant circulatory effector cells and play a vital role in driving or resolving inflammatory processes depending on their activation phenotype. We investigated and compared a panel of polarization protocols of blood-derived monocytes to achieve a stable, optimal and effective regimen for in vitro induction of immunosuppressive human macrophages, evaluating their surface receptor expression, cytokine profile, scavenging function and ability to suppress T-cell proliferation. Importantly, we assessed the effect of copolarization or secondary pro-inflammatory stimulation of a primary anti-inflammatory activation phenotype. A combination of IL-4/IL-10/TGF-β yielded a relatively stable and dominant immunosuppressive phenotype characterized by higher IL-10 production and down-regulated TNF-α, IL-6, CD86, CD274 and MHC II expression. Functionally, IL-4/IL-10/TGF-β-stimulated macrophages (M2) had a potent deactivating effect on a subsequent pro-inflammatory LPS/IFNγ-activated macrophage (M1) stimulation and significantly suppressed T-cell proliferation. Monocytes derived from patients with chronic inflammatory diseases could be induced to be anti-inflammatory using this protocol. Pre-differentiation with GM-CSF or M-CSF was further demonstrated to enhance final M1/M2 activation status. Our findings indicate a robust polarization protocol for generation of specific immunosuppressive human monocyte-derived macrophages.
authors: Applied Immunology & Immunotherapy, Department of Clinical Neuroscience, Center for Molecular Medicine, Karolinska University Hospital at Solna, Karolinska Institutet, Stockholm, Sweden.
authors: S
authors: S
authors: Mia
authors: A
authors: A
authors: Warnecke
authors: X-M
authors: XM
authors: Zhang
authors: V
authors: V
authors: Malmström
authors: R A
authors: RA
authors: Harris
mesh_terms: D051940:B7-2 Antigen / Q000378:metabolism; D060890:B7-H1 Antigen / Q000378:metabolism; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D049109:Cell Proliferation; D002478:Cells, Cultured; D016207:Cytokines / Q000378:metabolism; D000949:Histocompatibility Antigens Class II / Q000378:metabolism; D006801:Humans; D007108:Immune Tolerance; D007165:Immunosuppression Therapy / Q000379:methods*; D016753:Interleukin-10 / Q000276:immunology; D015847:Interleukin-4 / Q000276:immunology; D016173:Macrophage Colony-Stimulating Factor / Q000276:immunology; D008264:Macrophages / Q000276:immunology*; D009000:Monocytes / Q000276:immunology*; D010641:Phenotype; D013601:T-Lymphocytes / Q000276:immunology*; D016212:Transforming Growth Factor beta / Q000276:immunology
chemical_list: D051940:B7-2 Antigen; D060890:B7-H1 Antigen; D016207:Cytokines; D000949:Histocompatibility Antigens Class II; D016212:Transforming Growth Factor beta; D016753:Interleukin-10; D015847:Interleukin-4; D016173:Macrophage Colony-Stimulating Factor