TY - JOUR

T1 - Immunity by formulation design

T2 - induction of high CD8+ T-cell responses by poly (I:C) incorporated into the CAF01 adjuvant via a double emulsion method

AU - Nordly, Pernille Juul

AU - Rose, Fabrice

AU - Christensen, Dennis

AU - Nielsen, Hanne Mørck

AU - Andersen, Peter

AU - Agger, Else Marie

AU - Foged, Camilla

PY - 2011

Y1 - 2011

N2 - The combination of nucleic acid-based Toll-like receptor (TLR)-3 or TLR9 agonists and cationic liposomes constitutes an effective vaccine adjuvant approach for eliciting CD8+ T-cell responses. However, complexing cationic liposomes and oppositely charged oligonucleotides generally results in highly unstable and heterogeneous formulations with limited clinical applicability. The aim of this study was to design, formulate, and carefully characterize a stable CD8-inducing adjuvant based on the TLR3 ligand polyinosinic–polycytidylic acid [poly(I:C)] incorporated into a cationic adjuvant system (CAF01) composed of dimethyldioctadecylammonium (DDA) and trehalose 6,6′-dibehenate (TDB). For this purpose, a modified double emulsion solvent evaporation method was investigated for complexation of high amounts of anionic poly(I:C) to gel-state DDA/TDB liposomes. Addition of a volatile, water-miscible co-solvent (ethanol) to the outer water phase enabled preparation of colloidally stable liposomes, presumably by reducing the poly(I:C)-enhanced rigidity of the lipid bilayer. Cryo-transmission electron microscopy (TEM) revealed the formation of unilamellar as well as multilamellar liposomes, the latter suggesting that poly(I:C) is intercalated between the membrane bilayers in an onion-like structure. Finally, immunization of mice with the model antigen ovalbumin (OVA) and DDA/TDB/poly(I:C) liposomes induced a remarkably strong, antigen-specific CD8+ T-cell response, which was maintained for more than two months. Importantly, whereas injection of soluble poly(I:C) led to rapid production of the pro-inflammatory cytokines tumor necrosis factor (TNF)-α and interleukin (IL)-6 in serum, administration of poly(I:C) in complex with the cationic DDA/TDB liposomes prevented this non-specific systemic pro-inflammatory response. These data emphasize the importance of improving the quality of the vaccine formulation to indeed overcome some of the major obstacles for using CD8-inducing agents such as poly(I:C) in future subunit vaccines.

AB - The combination of nucleic acid-based Toll-like receptor (TLR)-3 or TLR9 agonists and cationic liposomes constitutes an effective vaccine adjuvant approach for eliciting CD8+ T-cell responses. However, complexing cationic liposomes and oppositely charged oligonucleotides generally results in highly unstable and heterogeneous formulations with limited clinical applicability. The aim of this study was to design, formulate, and carefully characterize a stable CD8-inducing adjuvant based on the TLR3 ligand polyinosinic–polycytidylic acid [poly(I:C)] incorporated into a cationic adjuvant system (CAF01) composed of dimethyldioctadecylammonium (DDA) and trehalose 6,6′-dibehenate (TDB). For this purpose, a modified double emulsion solvent evaporation method was investigated for complexation of high amounts of anionic poly(I:C) to gel-state DDA/TDB liposomes. Addition of a volatile, water-miscible co-solvent (ethanol) to the outer water phase enabled preparation of colloidally stable liposomes, presumably by reducing the poly(I:C)-enhanced rigidity of the lipid bilayer. Cryo-transmission electron microscopy (TEM) revealed the formation of unilamellar as well as multilamellar liposomes, the latter suggesting that poly(I:C) is intercalated between the membrane bilayers in an onion-like structure. Finally, immunization of mice with the model antigen ovalbumin (OVA) and DDA/TDB/poly(I:C) liposomes induced a remarkably strong, antigen-specific CD8+ T-cell response, which was maintained for more than two months. Importantly, whereas injection of soluble poly(I:C) led to rapid production of the pro-inflammatory cytokines tumor necrosis factor (TNF)-α and interleukin (IL)-6 in serum, administration of poly(I:C) in complex with the cationic DDA/TDB liposomes prevented this non-specific systemic pro-inflammatory response. These data emphasize the importance of improving the quality of the vaccine formulation to indeed overcome some of the major obstacles for using CD8-inducing agents such as poly(I:C) in future subunit vaccines.

KW - Former Faculty of Pharmaceutical Sciences

U2 - 10.1016/j.jconrel.2010.11.021

DO - 10.1016/j.jconrel.2010.11.021

M3 - Journal article

C2 - 21111765

VL - 150

SP - 307

EP - 317

JO - Journal of Controlled Release

JF - Journal of Controlled Release

SN - 0168-3659

IS - 3

ER -