Cross-presentation through langerin and DC-SIGN targeting requires different formulations of glycan-modified antigens

Dendritic cells (DCs) and Langerhans cells (LC) are professional antigen presenting cells (APCs) that initiate humoral and cellular immune responses. Targeted delivery of antigen towards DC- or LC-specific receptors enhances vaccine efficacy. In this study, we compared the efficiency of glycan-based antigen targeting to both the human DC-specific C-type lectin receptor (CLR) DC-SIGN and the LC-specific CLR langerin. Since DC-SIGN and langerin are able to recognize the difucosylated oligosaccharide Lewis Y (Le^Y), we prepared neoglycoconjugates bearing this glycan epitope to allow targeting of both lectins. Le^Y-modified liposomes, with an approximate diameter of 200 nm, were significantly endocytosed by DC-SIGN⁺ DCs and mediated efficient antigen presentation to CD4⁺ and CD8⁺ T cells. Surprisingly, although langerin bound to Le^Y-modified liposomes, LCs exposed to Le^Y-modified liposomes could not endocytose liposomes nor mediate antigen presentation to T cells. However, LCs mediated an enhanced cross-presentation when antigen was delivered through langerin using Le^Y-modified synthetic long peptides. In contrast, Le^Y-modified synthetic long peptides were recognized by DC-SIGN, but did not trigger antigen internalization nor antigen cross-presentation. These data demonstrate that langerin and DC-SIGN have different size requirements for antigen uptake. Although using glycans remains an interesting option in the design of anti-cancer vaccines targeting multiple CLRs, aspects such as molecule size and conformation need to be taken in consideration.