title: Biological evaluation of multivalent lewis X-MGL-1 interactions.
abstract: Myeloid C-type lectin receptors (CLRs) expressed by antigen-presenting cells are pattern-recognition receptors involved in the recognition of pathogens as well as of self-antigens. The interaction of carbohydrate ligands with a CLR can trigger immune responses. Although several CLR ligands are known, there is limited insight into CLR targeting by carbohydrate ligands. The weak affinity of lectin-carbohydrate interactions often renders multivalent carbohydrate presentation necessary. Here, we have analyzed the impact of multivalent presentation of the trisaccharide Lewis X (Le(X) ) epitope on its interaction with the CLR macrophage galactose-type lectin-1 (MGL-1). Glycan arrays, including N-glycan structures with terminal Le(X) , were prepared by enzymatic extension of immobilized synthetic core structures with two recombinant glycosyltransferases. Incubation of arrays with an MGL-1-hFc fusion protein showed up to tenfold increased binding to multiantennary N-glycans displaying Le(X) structures, compared to monovalent Le(X) trisaccharide. Multivalent presentation of Le(X) on the model antigen ovalbumin (OVA) led to increased cytokine production in a dendritic cell /T cell coculture system. Furthermore, immunization of mice with Le(X) -OVA conjugates modulated cytokine production and the humoral response, compared to OVA alone. This study provides insights into how multivalent carbohydrate-lectin interactions can be exploited to modulate immune responses.
authors.affiliation: Department of Biomolecular Systems, Max Planck Institute of Colloids and Interfaces, Am Mühlenberg 1, 14476 Potsdam (Germany); Institute for Chemistry and Biochemistry, Freie Universität Berlin, Arnimallee 22, 14195 Berlin (Germany).
authors.forename: Magdalena
authors.initials: M
authors.lastname: Eriksson
authors.forename: Sonia
authors.initials: S
authors.lastname: Serna
authors.forename: Maha
authors.initials: M
authors.lastname: Maglinao
authors.forename: Mark K
authors.initials: MK
authors.lastname: Schlegel
authors.forename: Peter H
authors.initials: PH
authors.lastname: Seeberger
authors.forename: Niels-Christian
authors.initials: NC
authors.lastname: Reichardt
authors.forename: Bernd
authors.initials: B
authors.lastname: Lepenies
mesh_terms: D000818:Animals; D001212:Asialoglycoproteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D002240:Carbohydrate Sequence; D002478:Cells, Cultured; D018920:Coculture Techniques; D016207:Cytokines / Q000378:metabolism; D003713:Dendritic Cells / Q000166:cytology / Q000276:immunology / Q000378:metabolism; D016695:Glycosyltransferases / Q000235:genetics / Q000378:metabolism; D006801:Humans; D056724:Immunity, Humoral; D007141:Immunoglobulin Fc Fragments / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D037181:Lectins, C-Type / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D016256:Lewis X Antigen / Q000737:chemistry* / Q000276:immunology / Q000378:metabolism; D008565:Membrane Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D008969:Molecular Sequence Data; D010047:Ovalbumin / Q000737:chemistry / Q000276:immunology; D011134:Polysaccharides / Q000032:analysis; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D013601:T-Lymphocytes / Q000166:cytology / Q000276:immunology / Q000378:metabolism
chemical_list: D001212:Asialoglycoproteins; C064720:Clec10a protein, mouse; D016207:Cytokines; D007141:Immunoglobulin Fc Fragments; D037181:Lectins, C-Type; D016256:Lewis X Antigen; D008565:Membrane Proteins; D011134:Polysaccharides; D011994:Recombinant Proteins; D010047:Ovalbumin; D016695:Glycosyltransferases
keywords: C-type lectins; antigens; carbohydrates; dendritic cells; multivalency; targeting