title: Glycosylation Changes Triggered by the Differentiation of Monocytic THP-1 Cell Line into Macrophages.
abstract: The human acute monocytic leukemia cell line THP-1 is widely used as an in vitro phagocytic cell model because it exhibits several immune properties similar to native monocyte-derived macrophages. In this study, we investigated the alteration of N- and O-linked glycans as well as glycosphingolipids, during THP-1 differentiation, combining mass spectrometry, flow cytometry, and quantitative real-time PCR. Mass spectrometry revealed that macrophage differentiation led to a marked upregulation of expression of GM3 ganglioside as well as an increase in complex-type structures, particularly triantennary glycans, occurring at the expense of high-mannose N-glycans. Moreover, we observed a slight decrease in the proportion of multifucosylated N-glycans and α2,6-sialylation. The uncovered changes in glycosylation correlated with variations of gene expression of relevant glycosyltransferases and glycosidases including sialyltransferases, β-N-acetylglucosaminyltransferases, fucosyltransferases, and neuraminidase. Furthermore, using flow cytometry and antibodies directed against glycan structures, we confirmed that the alteration of glycosylation occurs at the cell surface of THP-1 macrophage-like cells. Altogether, we established that macrophagic maturation of THP-1 induces dramatic modifications of the surface glycosylation pattern that may result in differential interaction of monocytic and macrophagic THP-1 with immune or bacterial lectins.
authors.affiliation: Univ. Lille , CNRS, UMR 8576 - UGSF - Unité de Glycobiologie Structurale et Fonctionnelle, F 59000 Lille, France.
authors.forename: Clément P
authors.initials: CP
authors.lastname: Delannoy
authors.affiliation: Univ. Lille , CNRS, UMR 8576 - UGSF - Unité de Glycobiologie Structurale et Fonctionnelle, F 59000 Lille, France.
authors.forename: Yoann
authors.initials: Y
authors.lastname: Rombouts
authors.affiliation: Univ. Lille , CNRS, UMR 8576 - UGSF - Unité de Glycobiologie Structurale et Fonctionnelle, F 59000 Lille, France.
authors.forename: Sophie
authors.initials: S
authors.lastname: Groux-Degroote
authors.affiliation: Center for Proteomics and Metabolomics, Leiden University Medical Center , 2300 RC Leiden, The Netherlands.
authors.forename: Stephanie
authors.initials: S
authors.lastname: Holst
authors.affiliation: Univ. Lille , CNRS, UMR 8576 - UGSF - Unité de Glycobiologie Structurale et Fonctionnelle, F 59000 Lille, France.
authors.forename: Bernadette
authors.initials: B
authors.lastname: Coddeville
authors.affiliation: Univ. Lille , CNRS, UMR 8576 - UGSF - Unité de Glycobiologie Structurale et Fonctionnelle, F 59000 Lille, France.
authors.forename: Anne
authors.initials: A
authors.lastname: Harduin-Lepers
authors.affiliation: Center for Proteomics and Metabolomics, Leiden University Medical Center , 2300 RC Leiden, The Netherlands.
authors.forename: Manfred
authors.initials: M
authors.lastname: Wuhrer
authors.affiliation: Univ. Lille , CNRS, UMR 8576 - UGSF - Unité de Glycobiologie Structurale et Fonctionnelle, F 59000 Lille, France.
authors.forename: Elisabeth
authors.initials: E
authors.lastname: Elass-Rochard
authors.affiliation: Univ. Lille , CNRS, UMR 8576 - UGSF - Unité de Glycobiologie Structurale et Fonctionnelle, F 59000 Lille, France.
authors.forename: Yann
authors.initials: Y
authors.lastname: Guérardel
mesh_terms: D002236:Carbohydrate Conformation; D002240:Carbohydrate Sequence; D002454:Cell Differentiation / Q000276:immunology*; D002460:Cell Line; D005647:Fucosyltransferases / Q000235:genetics / Q000276:immunology; D005679:G(M3) Ganglioside / Q000737:chemistry / Q000276:immunology; D005786:Gene Expression Regulation; D006028:Glycosphingolipids / Q000737:chemistry* / Q000276:immunology; D006031:Glycosylation; D016695:Glycosyltransferases / Q000235:genetics / Q000276:immunology; D006801:Humans; D008264:Macrophages / Q000737:chemistry* / Q000166:cytology / Q000276:immunology; D008358:Mannose / Q000737:chemistry / Q000276:immunology; D009000:Monocytes / Q000737:chemistry* / Q000166:cytology / Q000276:immunology; D017351:N-Acetylglucosaminyltransferases / Q000235:genetics / Q000276:immunology; D009439:Neuraminidase / Q000235:genetics / Q000276:immunology; D011134:Polysaccharides / Q000737:chemistry* / Q000276:immunology; D012794:Sialic Acids / Q000737:chemistry / Q000276:immunology; D012799:Sialyltransferases / Q000235:genetics / Q000276:immunology
chemical_list: D005679:G(M3) Ganglioside; D006028:Glycosphingolipids; D011134:Polysaccharides; D012794:Sialic Acids; D016695:Glycosyltransferases; D005647:Fucosyltransferases; D017351:N-Acetylglucosaminyltransferases; D012799:Sialyltransferases; D009439:Neuraminidase; D008358:Mannose
keywords: glycosphingolipids; glycosylation; glycosyltransferases; mass spectrometry; monocyte-to-macrophage differentiation