Altering the ligand specificity of DectiSomes.

DectiSomes are drug-loaded liposomes coated with pathogen receptors, such as the C-type lectins (CTL) Dectin-2 (D2) and Dectin-3 (D3, MCL). Floating on the surface of DectiSomes, the carbohydrate recognition domains (CRDs) of these CTLs form dimers that bind their cognate oligoglycan ligands. We have shown previously that amphotericin B (AmB)-loaded DectiSomes, D2-AmB-LLs, and D3-AmB-LLs, are effective at binding and killing diverse pathogenic fungi. The best-known ligands of Dectin-2 and Dectin-3 in the Candida albicans cell wall and exopolysaccharide matrix include a wide variety of oligomannans. When D2-AmB-LLs or D3-AmB-LLs were labeled in their lumen with complementary green and red fluorescent proteins, Venus and mCherry, they bound the same overlapping regions of oligoglycans in C. albicans colonies. By contrast, when D2-AmB-LLs and D3-AmB-LLs were labeled on their membrane surfaces with complementary pairs of the small fluorophores FITC and Rhodamine B or with Venus and mCherry, they bound mostly nonoverlapping sets of ligands. When the Dectin-2 and Dectin-3 proteins were labeled with the complementary pairs of FITC and Rhodamine, they also bound primarily distinct ligands. We proposed several models to explain these alterations in Dectin and DectiSome ligand specificity. These findings also raise important questions about the ligand binding properties of immuno-liposomes.