title: Preclinical profiling of the immunogenicity of a two-component subunit malaria vaccine candidate based on virosome technology.
abstract: Presentation of synthetic peptides on immunopotentiating reconstituted influenza virosomes is a promising technology for subunit vaccine development. An optimized virosomally delivered peptide representing 5 NPNA repeats of P. falciparum circumsporozoite protein is highly immunogenic in mice. Antibodies against this peptide (UK-39) inhibit sporozoite invasion of human hepatocytes. A second peptide (AMA49-C1) based on domain III of apical membrane antigen 1, induces antibodies that inhibit blood-stage parasite growth in vitro. Here we show a detailed pre-clinical profiling of these virosomally formulated peptides alone and in combination in mice and rabbits. Two immunizations with virosomally formulated UK-39 or AMA49-C1 were enough to elicit high titers of parasite cross-reactive antibodies in both species. A low dose of 10 microg UK-39 was enough to induce maximal titers in rabbits. Higher doses of peptide did not increase antibody titers. In contrast, AMA49-C1 induced higher antibody titers with 25 and 50 microg peptide. Combination of UK-39 and AMA49- C1 on separate virosomes did not have any negative effect on anti-peptide antibody titers in mice or rabbits. No MHC restriction was observed in the development of humoral responses in outbred rabbits with different immunogenetic backgrounds. All vaccine formulations were safe in toxicity studies in rabbits and rats. Taken together, low amounts of synthetic peptides delivered on virosomes induced high antibody titers in mice and rabbits. Moreover, different peptides could be combined without interfering with individual anti-peptide responses, augmenting the value of this system for the development of a multivalent malaria vaccine.
authors.affiliation: Molecular Immunology, Swiss Tropical Institute, Basel, Switzerland.
authors.forename: Shinji L
authors.initials: SL
authors.lastname: Okitsu
authors.forename: Markus S
authors.initials: MS
authors.lastname: Mueller
authors.forename: Mario
authors.initials: M
authors.lastname: Amacker
authors.forename: Denise
authors.initials: D
authors.lastname: Vogel
authors.forename: Nicole
authors.initials: N
authors.lastname: Westerfeld
authors.forename: John A
authors.initials: JA
authors.lastname: Robinson
authors.forename: Rinaldo
authors.initials: R
authors.lastname: Zurbriggen
authors.forename: Gerd
authors.initials: G
authors.lastname: Pluschke
mesh_terms: D000818:Animals; D000913:Antibodies, Protozoan / Q000097:blood*; D000953:Antigens, Protozoan / Q000737:chemistry / Q000276:immunology*; D003429:Cross Reactions; D004306:Dose-Response Relationship, Immunologic; D015195:Drug Design; D004912:Erythrocytes / Q000469:parasitology; D005260:Female; D007114:Immunization; D007074:Immunoglobulin G / Q000097:blood; D017780:Malaria Vaccines / Q000008:administration & dosage / Q000737:chemistry / Q000276:immunology*; D016778:Malaria, Falciparum / Q000276:immunology / Q000469:parasitology / Q000517:prevention & control; D008297:Male; D008565:Membrane Proteins / Q000138:chemical synthesis / Q000737:chemistry / Q000276:immunology*; D051379:Mice; D008807:Mice, Inbred BALB C; D010455:Peptides / Q000008:administration & dosage / Q000138:chemical synthesis / Q000737:chemistry / Q000276:immunology; D010963:Plasmodium falciparum / Q000276:immunology; D015800:Protozoan Proteins / Q000138:chemical synthesis / Q000737:chemistry / Q000276:immunology*; D051381:Rats; D017208:Rats, Wistar; D022701:Virosomes / Q000276:immunology*
chemical_list: D000913:Antibodies, Protozoan; D000953:Antigens, Protozoan; D007074:Immunoglobulin G; D017780:Malaria Vaccines; D008565:Membrane Proteins; D010455:Peptides; D015800:Protozoan Proteins; D022701:Virosomes; C090077:apical membrane antigen I, Plasmodium; C041595:circumsporozoite protein, Protozoan