Preparation and In Vitro Evaluation of Imiquimod Loaded Polylactide-based Micelles as Potential Vaccine Adjuvants
Résumé
Activation of immune cells through pattern recognition receptors (PRRs), such as Toll-like receptors (TLRs) or NOD-like receptors (NLRs), has been identified as a key issue in the development of new efficient vaccine adjuvants. We report here on the elaboration and immunostimulatory potential of polylactide (PLA)-based micelles core-loaded with imiquimod TLR7 ligand and able to be further surface-functionalized with antigenic protein (HIV-1 Gag p24) for antigen delivery purpose. Micelles prepared from poly(D,L-lactide)-b-poly(N-acryloxysuccinimide-co-N-vinylpyrrolidone) amphiphilic copolymer were incubated in the presence of imiquimod, leading to 1.2 wt% loading, and further conjugated to p24 antigen through reaction of p24 lysines and N-terminal amine with the N-succinimidyl pendant groups of the micelle corona. The impact of imiquimod encapsulation in the micelles on its immunostimulatory properties was investigated in vitro, by monitoring: (i) the NF-kappa B and mitogen-activated protein kinases (MAPK) pathways through experiments with RAW-Blue (TM) cells, a mouse macrophage cell line encoding an NF-kappa B/AP-1-inducible reporter construct; (ii) human dendritic cells (DCs) maturation markers by flow cytometry. RAW-Blue (TM) cells based experiments showed that imiquimod encapsulated in the micelles was much more efficient to activate the NF-kappa B and MAPK pathways than free imiquimod. Furthermore, encapsulated imiquimod was found to induce much higher maturation of DCs than the free analog. Finally, these immunostimulatory properties of the loaded imiquimod were shown to be conserved when the p24 antigen was coupled at the micelle surface. Taken together, these data regarding improved immunostimulatory efficiency suggest the strong potential of our micelle-based nano-system for vaccine delivery.