Conjugation‐Free Multilamellar Protein‐Lipid Hybrid Vesicles for Multifaceted Immune Responses

Abstract

Various lipid-based nanocarriers have been developed for the co-delivery of protein antigens with immunological adjuvants. However, their in vivo potency in vaccine delivery is limited by structural instability, which causes off-target delivery and low cross-presentation efficacies. Recent works employ covalent cross-linking to stabilize the lipid nanostructures, though the immunogenicity and side effects of chemically modified protein antigens and lipids can cause a long-lasting safety issue. Here robust "conjugation-free" multilamellar protein antigen-lipid hybrid nanovesicles (MPLVs) are introduced through the antigen-mediated self-assembly of unilamellar lipid vesicles for the co-delivery of protein antigens and immunologic adjuvants. The nanocarriers coated with monophosphoryl lipid A and hyaluronic acids elicit highly increase antigen-specific immune responses in vitro and in vivo. The MPLVs increase the generation of immunological surface markers and cytokines in mouse-derived bone-marrow dendritic cells compared to soluble antigens with adjuvants. Besides, the vaccination of mice with the MPLVs significantly increase the production of anti-antigen antibody and interferon-gamma via the activation of CD4 + and CD8 + T cells, respectively. These findings suggest that MPLVs can serve as a promising nanovaccine delivery platform for efficient antigen cross-presentation through the efficient co-delivery of protein antigens with adjuvants.

Highly Robust Multilamellar Lipid Vesicles Generated through Intervesicular Self-assembly Mediated by Hydrolyzed Collagen Peptides

“…The hybrid vesicles coated with hydrophobic moleculeconjugated polymers also improved long-term stable circulation [16,17]. Recently, we reported that some oppositely charged globular proteins can induce the structural changes of pre-formed charged lipid nanovesicles into multilamellar structures through polyionic interaction under certain conditions [18,19]. Our approach is similar to nucleic acid-lipid ionic complexation, where negatively charged nucleic acids are intercalated into the interstitial space of cationic lipid vesicles to generate closely packed arrays through spontaneous phase conversion [20][21][22].…”

“…SAXS analysis was performed at the 4C SAXS II beamline at the Pohang Accelerator Laboratory (Pohang, Republic of Korea) [18,19]. The lipid particles were detected under 0.734 of X-ray beam wavelength with 2 m of sample-to-detector distance (SDD) for 10 sec at 25 o C. The magnitude of the scattering vector (q = (4π/λ)sinθ, θ = scattering angle) was determined between 0.01 Å -1 and 0.3 Å -1 .…”

“…The charge neutralization was also observed as the zeta potential dramatically decreased from 52.9 ± 0.6 mV to 17.9 ± 1.4 mV (WR = 5.0) as LUVs were complexed with HCPs. The decreased zeta potential and particle size of MCLVs suggest the facile binding of HCPs onto the surface of DOTAP LUVs, resulting in significant morphological changes of LUVs during the formation of multilamellar structures as observed in the DNA-lipid and globular protein-lipid complexation [18,19]. Note that the simple coating of LUVs with HCPs cannot explain the shrinkage of LUVs.…”

Highly Robust Multilamellar Lipid Vesicles Generated through Intervesicular Self-Assembly Mediated by Hydrolyzed Collagen Peptides

“…SAXS analysis was performed at the 4C SAXS II beamline at the Pohang Accelerator Laboratory (Pohang, Republic of Korea). , The lipid particles were detected under 0.734 of X-ray beam wavelength with 2 m of sample-to-detector distance (SDD) for 10 s at 25 °C. The q value ( q = (4π/λ)­sin θ, θ = scattering angle) was determined between 0.01 and 0.3 Å –1 .…”

“…16,17 Recently, we reported that some oppositely charged globular proteins can induce the structural changes of preformed charged lipid nanovesicles into multilamellar structures through polyionic interaction under certain conditions. 18,19 Our approach is similar to nucleic acid−lipid ionic complexation, where negatively charged nucleic acids are hybridized with cationic liposomes to generate nanovesicles through spontaneous phase conversion. 20−22 However, protein−lipid complexation is technically more challenging as the precise control of the intermolecular interaction between proteins and the lipid membranes of nanovesicles is complicated because of the low and heterogeneous surface charge density of proteins.…”