title: Plasmodium vivax ookinete surface protein Pvs25 linked to cholera toxin B subunit induces potent transmission-blocking immunity by intranasal as well as subcutaneous immunization.
abstract: The nontoxic cholera toxin B subunit (CTB) was evaluated as a potential delivery molecule for the Plasmodium vivax ookinete surface protein, Pvs25. Recombinant Pvs25 was expressed as a secreted protein in the yeast Pichia pastoris, as a mixture of isoforms including multimers and the A and B monomers. The A isoform with the presumed native protein fold was the most abundant, accounting for more than 40% of all expressed protein. The molecularly uniform A isoform was chemically conjugated to CTB via its primary amines, and the fusion protein, retaining GM1-ganglioside affinity, was administered to BALB/c mice by the subcutaneous (s.c.) or intranasal (i.n.) route. Immunization of mice with conjugated Pvs25 without supplemental adjuvant induced antisera that specifically recognized P. vivax ookinetes in vitro. Furthermore, the antisera, when mixed with parasitized blood isolated from P. vivax patients from Thailand, was found to reduce parasite transmission to mosquitoes, conferring a 93 to 98% (s.c.) or a 73 to 88% (i.n.) decrease in oocyst number. Unconjugated Pvs25 alone conferred only a 23 to 60% (s.c.) or a 0 to 6% (i.n.) decrease in oocyst number. Coadministration of extraneous adjuvants, however, further enhanced the vaccine efficacy up to complete blockade. Taken together, we conclude that a weakly immunogenic Pvs25 by itself, when linked to CTB, transforms into a potent transmission-blocking antigen in both i.n. and s.c. routes. In addition, the present study is, to the best of our knowledge, the first demonstration of the immune potentiating function of CTB for a vaccine antigen delivered by the s.c. route.
authors.affiliation: Molecular Microbiology Group, Department of Tropical Infectious Diseases, COMB, Tropical Biosphere Research Center, University of the Ryukyus, 1 Senbaru, Nishihara, Okinawa 903-0213, Japan.
authors.forename: Takeshi
authors.initials: T
authors.lastname: Miyata
authors.forename: Tetsuya
authors.initials: T
authors.lastname: Harakuni
authors.forename: Takafumi
authors.initials: T
authors.lastname: Tsuboi
authors.forename: Jetsumon
authors.initials: J
authors.lastname: Sattabongkot
authors.forename: Hideyasu
authors.initials: H
authors.lastname: Kohama
authors.forename: Mayumi
authors.initials: M
authors.lastname: Tachibana
authors.forename: Goro
authors.initials: G
authors.lastname: Matsuzaki
authors.forename: Motomi
authors.initials: M
authors.lastname: Torii
authors.forename: Takeshi
authors.initials: T
authors.lastname: Arakawa
mesh_terms: D000276:Adjuvants, Immunologic / Q000008:administration & dosage*; D000281:Administration, Intranasal; D000818:Animals; D000852:Anopheles; D000953:Antigens, Protozoan / Q000276:immunology*; D000954:Antigens, Surface / Q000276:immunology*; D002772:Cholera Toxin / Q000008:administration & dosage*; D005260:Female; D006801:Humans; D007279:Injections, Subcutaneous; D017780:Malaria Vaccines / Q000008:administration & dosage / Q000276:immunology*; D016780:Malaria, Vivax / Q000517:prevention & control / Q000635:transmission*; D051379:Mice; D008807:Mice, Inbred BALB C; D010966:Plasmodium vivax / Q000276:immunology*; D014614:Vaccines, Synthetic / Q000276:immunology
chemical_list: D000276:Adjuvants, Immunologic; D000953:Antigens, Protozoan; D000954:Antigens, Surface; D017780:Malaria Vaccines; C121473:Pvs25 protein, P vivax; D014614:Vaccines, Synthetic; D002772:Cholera Toxin