title: Immunogenicity, protective efficacy and mechanism of novel CCS adjuvanted influenza vaccine.
abstract: We optimized the immunogenicity of adjuvanted seasonal influenza vaccine based on commercial split influenza virus as an antigen (hemagglutinin = HA) and on a novel polycationic liposome as a potent adjuvant and efficient antigen carrier (CCS/C-HA vaccine). The vaccine was characterized physicochemically, and the mechanism of action of CCS/C as antigen carrier and adjuvant was studied. The optimized CCS/C-HA split virus vaccine, when administered intramuscularly (i.m.), is significantly more immunogenic in mice, rats and ferrets than split virus HA vaccine alone, and it provides for protective immunity in ferrets and mice against live virus challenge that exceeds the degree of efficacy of the split virus vaccine. Similar adjuvant effects of optimized CCS/C are also observed in mice for H1N1 swine influenza antigen. The CCS/C-HA vaccine enhances immune responses via the Th1 and Th2 pathways, and it increases both the humoral responses and the production of IL-2 and IFN-γ but not of the pro-inflammatory factor TNFα. In mice, levels of CD4(+) and CD8(+) T-cells and of MHC II and CD40 co-stimulatory molecules are also elevated. Structure-function relationship studies of the CCS molecule as an adjuvant/carrier show that replacing the saturated palmitoyl acyl chain with the mono-unsaturated oleoyl (C18:1) chain affects neither size distribution and zeta potential nor immune responses in mice. However, replacing the polyalkylamine head group spermine (having two secondary amines) with spermidine (having only one secondary amine) reduces the enhancement of the immune response by ∼ 50%, while polyalkylamines by themselves are ineffective in improving the immunogenicity over the commercial HA vaccine. This highlights the importance of the particulate nature of the carrier and the polyalkylamine secondary amines in the enhancement of the immune responses against seasonal influenza. Altogether, our results suggest that the CCS/C polycationic liposomes combine the activities of a potent adjuvant and efficient carrier of seasonal and swine flu vaccines and support further development of the CCS/C-HA vaccine.
authors.affiliation: Laboratory of Membrane and Liposome Research, Institute for Medical Research Israel-Canada, The Hebrew University-Hadassah Medical School, PO Box 12272, Jerusalem 91120, Israel.
authors.forename: Orli
authors.initials: O
authors.lastname: Even-Or
authors.forename: Sarit
authors.initials: S
authors.lastname: Samira
authors.forename: Eli
authors.initials: E
authors.lastname: Rochlin
authors.forename: Shobana
authors.initials: S
authors.lastname: Balasingam
authors.forename: Alex J
authors.initials: AJ
authors.lastname: Mann
authors.forename: Rob
authors.initials: R
authors.lastname: Lambkin-Williams
authors.forename: Jack
authors.initials: J
authors.lastname: Spira
authors.forename: Itzhak
authors.initials: I
authors.lastname: Goldwaser
authors.forename: Ronald
authors.initials: R
authors.lastname: Ellis
authors.forename: Yechezkel
authors.initials: Y
authors.lastname: Barenholz
mesh_terms: D000276:Adjuvants, Immunologic / Q000008:administration & dosage* / Q000737:chemistry*; D000818:Animals; D000956:Antigens, Viral / Q000276:immunology; D016207:Cytokines / Q000276:immunology; D005260:Female; D005289:Ferrets; D019267:Hemagglutinin Glycoproteins, Influenza Virus / Q000276:immunology; D007111:Immunity, Cellular; D056724:Immunity, Humoral; D007252:Influenza Vaccines / Q000008:administration & dosage / Q000276:immunology*; D007273:Injections, Intramuscular; D008081:Liposomes / Q000276:immunology*; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008809:Mice, Inbred C3H; D009976:Orthomyxoviridae Infections / Q000276:immunology / Q000517:prevention & control*; D051381:Rats; D017207:Rats, Sprague-Dawley
chemical_list: D000276:Adjuvants, Immunologic; D000956:Antigens, Viral; D016207:Cytokines; D019267:Hemagglutinin Glycoproteins, Influenza Virus; D007252:Influenza Vaccines; D008081:Liposomes