title: Differential constitutive and cytokine-modulated expression of human Toll-like receptors in primary neutrophils, monocytes, and macrophages.
abstract: Human Toll-like receptors (TLRs) comprise a family of proteins that recognizes pathogen-associated molecular patterns (PAMPs) and initiates host innate immune responses. Neutrophils, monocytes, and macrophages are critical cellular components of the human innate immune system. Proinflammatory cytokines, such as granulocyte colony-stimulating factor (G-CSF), granulocyte-macrophage colony-stimulating factor (GM-CSF), macrophage colony-stimulating factor (M-CSF), and interferon-gamma (IFN-gamma), have been shown to up-regulate microbicidal activity in these effector cells of innate immunity. Currently, the cellular and molecular mechanisms responsible for these effects are not completely understood. We hypothesized that these cytokines may up-regulate TLR expression as a mechanism to facilitate microbial recognition and augment the innate immune response. Using quantitative realtime rt-PCR technology, we examined constitutive expression of TLR2, TLR4, TLR5, and TLR9 mRNA and the effects of G-CSF, GM-CSF, M-CSF, and IFN-gamma on TLR mRNA expression in purified populations of normal human neutrophils, monocytes, and monocyte-derived macrophages. Relative constitutive expression of TLR2, TLR4, and TLR9 was similar in neutrophils and monocytes. Constitutive expression of TLR5 was less in neutrophils compared to monocytes. Constitutive expression of TLR4 was greater and that of TLR9 lower in monocyte-derived macrophages compared to monocytes. Of the cytokines examined, IFN-gamma and GM-CSF caused the greatest effects on TLR expression. IFN- gamma up-regulated TLR2 and TLR4 in neutrophils and monocytes. GM-CSF up-regulated expression of TLR2 and TLR4 in neutrophils and TLR2 in monocytes. TLR5 was down-regulated by inflammatory cytokines in monocytes. These results suggest a potential role for IFN- gamma and/or GM-CSF as therapeutic immunomodulators of the host defense to infection.
authors.affiliation: Departments of Medicine, University of Washington, Seattle, WA 98195, USA.
authors.forename: D Shane
authors.initials: DS
authors.lastname: O'Mahony
authors.forename: Uyenvy
authors.initials: U
authors.lastname: Pham
authors.forename: Ramesh
authors.initials: R
authors.lastname: Iyer
authors.forename: Thomas R
authors.initials: TR
authors.lastname: Hawn
authors.forename: W Conrad
authors.initials: WC
authors.lastname: Liles
mesh_terms: D018929:Cell Culture Techniques; D002478:Cells, Cultured; D016207:Cytokines / Q000235:genetics / Q000502:physiology*; D005786:Gene Expression Regulation / Q000276:immunology; D016179:Granulocyte Colony-Stimulating Factor / Q000235:genetics / Q000502:physiology; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000235:genetics / Q000502:physiology; D006801:Humans; D007371:Interferon-gamma / Q000235:genetics / Q000378:metabolism / Q000502:physiology; D008262:Macrophage Activation; D016173:Macrophage Colony-Stimulating Factor / Q000235:genetics / Q000502:physiology; D008264:Macrophages / Q000276:immunology / Q000502:physiology*; D009000:Monocytes / Q000276:immunology / Q000502:physiology*; D018375:Neutrophil Activation; D009504:Neutrophils / Q000276:immunology / Q000502:physiology*; D012333:RNA, Messenger / Q000378:metabolism; D011994:Recombinant Proteins / Q000378:metabolism; D051195:Toll-Like Receptor 2 / Q000378:metabolism / Q000502:physiology; D051197:Toll-Like Receptor 4 / Q000378:metabolism / Q000502:physiology; D051198:Toll-Like Receptor 5 / Q000378:metabolism / Q000502:physiology; D051217:Toll-Like Receptor 9 / Q000378:metabolism / Q000502:physiology; D051193:Toll-Like Receptors / Q000378:metabolism* / Q000502:physiology
chemical_list: D016207:Cytokines; D012333:RNA, Messenger; D011994:Recombinant Proteins; D051195:Toll-Like Receptor 2; D051197:Toll-Like Receptor 4; D051198:Toll-Like Receptor 5; D051217:Toll-Like Receptor 9; D051193:Toll-Like Receptors; D016179:Granulocyte Colony-Stimulating Factor; D016173:Macrophage Colony-Stimulating Factor; D007371:Interferon-gamma; D016178:Granulocyte-Macrophage Colony-Stimulating Factor