Targeting Langerhans cells using a modular mannosylated nucleic acid-based vaccine platform

The skin is a diverse reservoir of immune cells with strong potential for immunotherapeutic delivery. Langerhans cells in the epidermis are antigen-presenting cells, accessible for vaccination using carbohydrate-conjugated therapeutics targeting their endocytic lectin receptor, Langerin. As carbohydrate-lectin binding is highly dependent on valency, scaffolds that enable control over ligand spacing and stoichiometry are instrumental in enhancing receptor binding and selectivity. Here we utilized a self-assembled nucleic acid-based Holliday Junction scaffold, fully modified for nuclease protection and with a well-defined carbohydrate arrangement to optimize drug delivery to Langerhans cells. In vitro screening with Langerin-expressing cells revealed that mannosylated Holliday Junctions showed the strongest binding. This was confirmed in human epidermal cell suspensions, demonstrating specificity and valency-driven interactions. Topical administration of mannosylated scaffolds on skin explants enabled effective targeting of epidermal Langerhans cells. Finally, in an antigen-presentation assay, in vitro differentiated Langerhans cells loaded with mannosylated and peptide-conjugated scaffolds significantly enhanced T cell activation. Overall, our study presents a promising nucleic acid-based platform for precise Langerhans cell targeting and drug delivery, with broad potential for skin-directed immunotherapies.