The use of dendritic cells (DC) for the development of therapeutic
cancer vaccines is attractive because of their unique ability to present
tumor epitopes via the MHC class I pathway to induce cytotoxic CD8+ T lymphocyte responses. C-Type membrane
lectins,
DC-SIGN and the
mannose receptor (MR), present on the DC surface, recognize
oligosaccharides containing
mannose and/or
fucose and mediate
sugar-specific endocytosis of synthetic oligolysine-based glycoclusters. We therefore asked whether a glycotargeting approach could be used to induce uptake and presentation of
tumor antigens by DC. To this end, we designed and synthesized glycocluster conjugates containing a CD8+
epitope of the
Melan-A/Mart-1
melanoma antigen. These glycocluster-
Melan-A conjugates were obtained by coupling glycosynthons: oligosaccharyl-pyroglutamyl-
beta-alanine derivatives containing either
disaccharides, a dimannoside (Manalpha-6Man) or lactoside, or a Lewis
oligosaccharide, to
Melan-A 16-40
peptide comprising the 26-35
HLA-A2 restricted
T cell epitope, extended with an oligolysine stretch at the C-terminal end. We showed by confocal microscopy and flow cytometry that fluorescent-labeled
Melan-A glycoclusters containing either dimannoside or Lewis
oligosaccharide were taken up by DC and concentrated in acidic vesicles; conversely lactoside
glycopeptides were not at all taken up. Furthermore, using surface plasmon resonance, we showed that dimannoside and Lewis-
Melan-A conjugates bind MR and
DC-SIGN with high affinity. DC loaded with these conjugates, but not with the
lactose-
Melan-A conjugate, led to an efficient presentation of the
Melan-A epitope eliciting a CD8+ T-lymphocyte response. These data suggest that synthetically designed glycocluster-
tumor antigen conjugates may induce
antigen cross-presentation by DC and represent a promising tool for the development of
tumor vaccines.