title: Trehalose diester glycolipids are superior to the monoesters in binding to Mincle, activation of macrophages in vitro and adjuvant activity in vivo.
abstract: The T-cell adjuvanticity of mycobacterial cord factor trehalose 6,6'-dimycolate (TDM) is well established. The identification of the C-type lectin Mincle on innate immune cells as the receptor for TDM and its synthetic analogue trehalose 6,6'-dibehenate (TDB) has raised interest in development of synthetic Mincle ligands as novel adjuvants. Trehalose mono- (TMXs) and diesters (TDXs) with symmetrically shortened acyl chains [denoted by X: arachidate (A), stearate (S), palmitate (P), and myristate (M)] were tested. Upon stimulation of murine macrophages, G-CSF secretion and NO production were strongly augmented by all TDXs tested, in a wide concentration range. In contrast, the TMXs triggered macrophage activation only at high concentrations. Macrophage activation by all TDXs required Mincle, but was independent of MyD88. The superior capacity of TDXs for activating macrophages was paralleled by direct binding of TDXs, but not of TMXs, to a Mincle-Fc fusion protein. Insertion of a short polyethylene glycol between the sugar and acyl chain in TDS reduced Mincle-binding and macrophage activation. Immunization of mice with cationic liposomes containing the analogues demonstrated the superior adjuvant activity of trehalose diesters. Overall, immune activation in vitro and in vivo by trehalose esters of simple fatty acids requires two acyl chains of length and involves Mincle.
authors.affiliation: Institute of Clinical Microbiology, Immunology and Hygiene, University Hospital Erlangen, Friedrich Alexander Universität Erlangen-Nürnberg, Berlin, Germany.
authors.forename: Alexandra
authors.initials: A
authors.lastname: Huber
authors.affiliation: Department of Pharmacy, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark Department of Drug Design and Pharmacology, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark Department of Infectious Disease Immunology, Vaccine Adjuvant Research, Statens Serum Institut, Copenhagen, Denmark.
authors.forename: Rie S
authors.initials: RS
authors.lastname: Kallerup
authors.affiliation: Department of Infectious Disease Immunology, Vaccine Adjuvant Research, Statens Serum Institut, Copenhagen, Denmark.
authors.forename: Karen S
authors.initials: KS
authors.lastname: Korsholm
authors.affiliation: Department of Drug Design and Pharmacology, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark.
authors.forename: Henrik
authors.initials: H
authors.lastname: Franzyk
authors.affiliation: Department of Biomolecular Systems, Max Planck Institute of Colloids and Interfaces, Potsdam, Germany Institute of Chemistry and Biochemistry, Department of Biology, Chemistry and Pharmacy, Freie Universität Berlin, Berlin, Germany Research Center for Emerging Infections and Zoonoses, University of Veterinary Medicine, Hannover, Germany.
authors.forename: Bernd
authors.initials: B
authors.lastname: Lepenies
authors.affiliation: Department of Infectious Disease Immunology, Vaccine Adjuvant Research, Statens Serum Institut, Copenhagen, Denmark.
authors.forename: Dennis
authors.initials: D
authors.lastname: Christensen
authors.affiliation: Department of Pharmacy, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark camilla.foged@sund.ku.dk.
authors.forename: Camilla
authors.initials: C
authors.lastname: Foged
authors.affiliation: Institute of Clinical Microbiology, Immunology and Hygiene, University Hospital Erlangen, Friedrich Alexander Universität Erlangen-Nürnberg, Berlin, Germany roland.lang@uk-erlangen.de.
authors.forename: Roland
authors.initials: R
authors.lastname: Lang
mesh_terms: D000276:Adjuvants, Immunologic / Q000008:administration & dosage* / Q000737:chemistry; D000818:Animals; D002478:Cells, Cultured; D006017:Glycolipids / Q000737:chemistry; D016179:Granulocyte Colony-Stimulating Factor / Q000276:immunology; D006801:Humans; D007113:Immunity, Innate / Q000187:drug effects; D016219:Immunotherapy, Adoptive / Q000379:methods*; D037181:Lectins, C-Type / Q000235:genetics / Q000378:metabolism*; D008262:Macrophage Activation / Q000187:drug effects; D008264:Macrophages / Q000502:physiology*; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D010703:Phorbol Esters / Q000737:chemistry; D011485:Protein Binding; D013601:T-Lymphocytes / Q000276:immunology / Q000637:transplantation*; D014199:Trehalose / Q000008:administration & dosage* / Q000031:analogs & derivatives / Q000737:chemistry
chemical_list: D000276:Adjuvants, Immunologic; C113583:Clecsf8 protein, mouse; D006017:Glycolipids; D037181:Lectins, C-Type; D008565:Membrane Proteins; D010703:Phorbol Esters; D016179:Granulocyte Colony-Stimulating Factor; D014199:Trehalose
keywords: C-type lectin receptor; Mincle; Vaccine adjuvant; trehalose dimycolate