Influence of Core β-1,2-Xylosylation on Glycoprotein Recognition by Murine C-type Lectin Receptors and Its Impact on Dendritic Cell Targeting.
Targeting antigens to dendritic cell subsets is a promising strategy to enhance the efficacy of vaccines. C-type lectin receptors (CLRs) expressed by dendritic cells are particularly attractive candidates since CLR engagement may promote cell uptake and may further stimulate antigen presentation and subsequent T cell activation. While most previous approaches have involved antibody-mediated CLR-targeting, glycan-based CLR targeting has become more and more attractive in recent years. In the present study, we show that small structural glycan modifications may markedly influence CLR recognition, dendritic cell targeting, and subsequent T cell activation. A biantennary N-glycan (G0) and its analogous O-2 core xylosylated N-glycan (XG0) were synthesized, covalently conjugated to the model antigen ovalbumin, and analyzed for binding to a set of murine CLR-Fc fusion proteins using lectin microarray. To evaluate whether the differential binding of G0 and XG0 to CLRs impacted dendritic cell targeting, uptake studies using murine dendritic cells were performed. Finally, effects of the ovalbumin glycoconjugates on T cell activation were measured in a dendritic cell/T cell cocultivation assay. Our results highlight the utility of glycan-based dendritic cell targeting and demonstrate that small structural differences may have a major impact on dendritic cell targeting efficacy.
CIC biomaGUNE , Glycotechnology Laboratory, Paseo Miramón 182, 20009 San Sebastian, Spain.
Katarzyna
K
Brzezicka
Max Planck Institute of Colloids and Interfaces , Department of Biomolecular Systems, Am Mühlenberg 1, 14476 Potsdam, Germany.
Uwe
U
Vogel
CIC biomaGUNE , Glycotechnology Laboratory, Paseo Miramón 182, 20009 San Sebastian, Spain.
Sonia
S
Serna
Max Planck Institute of Colloids and Interfaces , Department of Biomolecular Systems, Am Mühlenberg 1, 14476 Potsdam, Germany.|Freie Universität Berlin , Institute of Chemistry and Biochemistry, Arnimallee 22, 14195 Berlin, Germany.|University of Veterinary Medicine Hannover , Immunology Group, Research Center of Emerging Infections and Zoonoses (RIZ), Bünteweg 17, 30559 Hannover, Germany.
Timo
T
Johannssen
Max Planck Institute of Colloids and Interfaces , Department of Biomolecular Systems, Am Mühlenberg 1, 14476 Potsdam, Germany.|Freie Universität Berlin , Institute of Chemistry and Biochemistry, Arnimallee 22, 14195 Berlin, Germany.|University of Veterinary Medicine Hannover , Immunology Group, Research Center of Emerging Infections and Zoonoses (RIZ), Bünteweg 17, 30559 Hannover, Germany.
Bernd
B
Lepenies
CIC biomaGUNE , Glycotechnology Laboratory, Paseo Miramón 182, 20009 San Sebastian, Spain.|CIBER-BBN , Paseo Miramón 182, 20009 San Sebastian, Spain.
Niels-Christian
NC
Reichardt
D000818:Animals; D003713:Dendritic Cells / Q000276:immunology*; D006023:Glycoproteins / Q000378:metabolism*; D037181:Lectins, C-Type / Q000378:metabolism*; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D010047:Ovalbumin / Q000378:metabolism; D011485:Protein Binding; D011975:Receptors, Mitogen / Q000378:metabolism*; D014994:Xylose / Q000378:metabolism*
D006023:Glycoproteins; D037181:Lectins, C-Type; D011975:Receptors, Mitogen; D010047:Ovalbumin; D014994:Xylose