title: Isoform localization of Dectin-1 regulates the signaling quality of anti-fungal immunity.
abstract: Dectin-1 is recognized as a major receptor for fungal ß-glucans and contributes to anti-fungal immunity. Human monocyte populations express Dectin-1 isoforms A and B, which differ by the presence of a stalk region and its N-linked glycosylation site. Here, we analyzed the expression of both isoforms in human monocyte-derived cells. The cellular localization on cell lines stably expressing either Dectin-1 isoform A or B was studied by flow cytometry and confocal laser scanning microscopy. Intracellular protein signaling and cytokine production were analyzed by immunoblotting and cytometric bead array, respectively. Monocyte-derived cells showed cell type-specific expression of the two isoforms. Glycosylated Dectin-1 isoform A was predominantly localized at the cell surface, non-glycosylated isoform B was retained intracellularly. Inhibition of glycosylation resulted in efficient abrogation of cell surface expression of isoform A. Signaling quality following Dectin-1 stimulation was reduced in isoform B cells. Differential isoform specific cytokine secretion was observed by cytometric bead array. We show here that n-glycosylation of Dectin-1 is crucial for its cell surface expression and consequently signal transduction. Taken together, unique cytokine secretion and varying expression levels of human Dectin-1 isoforms on monocyte-derived cells may indicate distinct isoform usage as a cell type-specific mechanism of regulating anti-fungal immunity.
authors.affiliation: Klinik für Innere Medizin II, Abteilung für Hämatologie und Internistische Onkologie, Universitätsklinikum Jena, Jena, Germany.
authors.forename: Mike
authors.initials: M
authors.lastname: Fischer
authors.affiliation: Institut für Molekulare Zellbiologie, CMB, Universitätsklinikum Jena, Jena, Germany.
authors.forename: Jörg P
authors.initials: JP
authors.lastname: Müller
authors.affiliation: Klinik für Innere Medizin II, Abteilung für Hämatologie und Internistische Onkologie, Universitätsklinikum Jena, Jena, Germany.
authors.forename: Bärbel
authors.initials: B
authors.lastname: Spies-Weisshart
authors.affiliation: Klinik für Innere Medizin II, Abteilung für Hämatologie und Internistische Onkologie, Universitätsklinikum Jena, Jena, Germany.
authors.forename: Christine
authors.initials: C
authors.lastname: Gräfe
authors.affiliation: Septomics Research Center, Friedrich Schiller University Jena, Leibniz Institute for Natural Product Research and Infection Biology, Hans-Knoell-Institute, Jena, Germany.
authors.forename: Oliver
authors.initials: O
authors.lastname: Kurzai
authors.affiliation: Septomics Research Center, Friedrich Schiller University Jena, Leibniz Institute for Natural Product Research and Infection Biology, Hans-Knoell-Institute, Jena, Germany.
authors.forename: Kerstin
authors.initials: K
authors.lastname: Hünniger
authors.affiliation: Klinik für Innere Medizin II, Abteilung für Hämatologie und Internistische Onkologie, Universitätsklinikum Jena, Jena, Germany.
authors.forename: Andreas
authors.initials: A
authors.lastname: Hochhaus
authors.affiliation: Klinik für Innere Medizin II, Abteilung für Hämatologie und Internistische Onkologie, Universitätsklinikum Jena, Jena, Germany.
authors.forename: Sebastian
authors.initials: S
authors.lastname: Scholl
authors.affiliation: Klinik für Innere Medizin II, Abteilung für Hämatologie und Internistische Onkologie, Universitätsklinikum Jena, Jena, Germany.
authors.forename: Ulf
authors.identifier: 0000-0001-7455-8988
authors.initials: U
authors.lastname: Schnetzke
mesh_terms: D002460:Cell Line; D016207:Cytokines / Q000096:biosynthesis / Q000276:immunology; D005434:Flow Cytometry; D006031:Glycosylation; D006801:Humans; D047908:Intracellular Signaling Peptides and Proteins / Q000378:metabolism; D037181:Lectins, C-Type / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D008264:Macrophages / Q000276:immunology; D008565:Membrane Proteins / Q000235:genetics / Q000276:immunology; D018613:Microscopy, Confocal; D009000:Monocytes / Q000276:immunology / Q000378:metabolism* / Q000502:physiology; D009181:Mycoses / Q000276:immunology* / Q000378:metabolism; D020033:Protein Isoforms / Q000737:chemistry / Q000378:metabolism; D015398:Signal Transduction; D000099181:Dectin-1
chemical_list: D016207:Cytokines; D047908:Intracellular Signaling Peptides and Proteins; D037181:Lectins, C-Type; D008565:Membrane Proteins; D020033:Protein Isoforms; C546929:CLEC7A protein, human; D000099181:Dectin-1
keywords: Cytokine secretion; Dectin-1 isoforms; Fungal diseases; Immune responses; Infectious diseases