Coadsorption of HIV-1 p24 and gp120 proteins to surfactant-free anionic PLA nanoparticles preserves antigenicity and immunogenicity.
Biodegradable micro- or nanoparticles with surface adsorbed antigens represent a promising method for in vivo delivery of vaccines. Most vaccines, licensed or under development, are based on combined delivery of multiple antigens. Thus, we investigated the feasibility of combining two vaccine antigens, HIV-1 p24 and gp120 proteins, on the surface of surfactant-free anionic PLA nanoparticles obtained by an improved solvent diffusion method. The analysis of adsorption isotherms has shown that both proteins had similar and high affinities for the nanoparticles. Coadsorption of p24 and gp120 onto the same PLA particle was evidenced by sandwich ELISA, using antibodies directed against one protein for particle capture and the other one for detection. To assess structural integrity, the antigenicity of free and PLA-adsorbed antigens was compared by competition ELISA, using a set of 6 anti-p24 and 7 anti-gp120 antibodies, as well as soluble CD4. The antigenicity of proteins on the nanoparticle surface was well preserved, adsorbed either individually or in combination. Furthermore, both antigens maintained their immunogenicity, since high antibody titres (10(6) for p24 and 10(5) for gp120) were elicited in mice with monovalent and divalent PLA formulations. Taken together our results show that development of multivalent vaccines based on anionic PLA nanoparticles is possible. Moreover, coadsorption of a ligand for cell-specific targeting or of an immunostimulatory molecule will further extend the field of application of delivery systems based on charged micro- and nanoparticles.
Lamalle-Bernard
Delphine
D
FRE2736 CNRS-bioMérieux, IFR128 BioSciences Lyon-Gerland, Tour CERVI, 21, Avenue Tony Garnier, F-69365 Lyon cedex 07, France.
Munier
Séverine
S
Compagnon
Christelle
C
Charles
Marie-Hélène
MH
Kalyanaraman
Vaniambadi S
VS
Delair
Thierry
T
Verrier
Bernard
B
Ataman-Onal
Yasemin
Y
D016915:AIDS Vaccines / Q000008:administration & dosage* / Q000276:immunology*; D000327:Adsorption; D000818:Animals; D000838:Anions; D000918:Antibody Specificity; D001667:Binding, Competitive; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000276:immunology; D004926:Escherichia coli / Q000235:genetics; D005260:Female; D016655:HIV Core Protein p24 / Q000008:administration & dosage* / Q000276:immunology*; D015699:HIV Envelope Protein gp120 / Q000008:administration & dosage* / Q000276:immunology*; D007114:Immunization; D019344:Lactic Acid; D051379:Mice; D008807:Mice, Inbred BALB C; D008970:Molecular Weight; D049329:Nanostructures; D010316:Particle Size; D011100:Polyglycolic Acid; D000077182:Polylactic Acid-Polyglycolic Acid Copolymer; D011108:Polymers; D011994:Recombinant Proteins / Q000008:administration & dosage / Q000276:immunology; D013816:Thermodynamics
D016915:AIDS Vaccines; D000838:Anions; D000939:Epitopes; D016655:HIV Core Protein p24; D015699:HIV Envelope Protein gp120; D011108:Polymers; D011994:Recombinant Proteins; D000077182:Polylactic Acid-Polyglycolic Acid Copolymer; D011100:Polyglycolic Acid; D019344:Lactic Acid