title: Saccharomyces cerevisiae-secreted fusion proteins Pfs25 and Pfs28 elicit potent Plasmodium falciparum transmission-blocking antibodies in mice.
abstract: Transmission-blocking vaccines based on sexual-stage surface antigens of Plasmodium falciparum may assist in the control of this lethal form of human malaria. Two vaccine candidates, Pfs25 and Pfs28, were produced as single recombinant fusion proteins. The 39-kDa chimeric proteins, having a C-terminal His6 tag, were secreted by Saccharomyces cerevisiae, using the prepro-alpha-factor leader sequence. Pfs25-28 fusion proteins were significantly more potent than either Pfs25 or Pfs28 alone in eliciting antibodies in mice that blocked oocyst development in Anopheles freeborni mosquitoes: complete inhibition of oocyst development in the mosquito midgut was achieved with fewer vaccinations, at a lower dose, and for a longer duration than with either Pfs25 or Pfs28 alone. Increased antigen-specific immunoglobulin G titers and highly significant lymphoproliferative stimulation by Pfs28-containing antigens suggest the presence of an immunodominant helper T-cell epitope in the Pfs28 portion of the fusion proteins. This epitope may be responsible for the enhanced humoral response to both Pfs25 and Pfs28 antigens. Protein production of the fusion protein was improved 12-fold by converting Pfs28 codons to yeast-preferred codons (TBV28), using a modified ADH2 promoter and incorporating a (Glu-Ala)2 repeat after the Kex2 cleavage site.
authors.affiliation: Laboratory of Parasitic Diseases, National Institutes of Health, Bethesda, Maryland 20892-0425, USA.
authors.forename: M M
authors.initials: MM
authors.lastname: Gozar
authors.forename: V L
authors.initials: VL
authors.lastname: Price
authors.forename: D C
authors.initials: DC
authors.lastname: Kaslow
mesh_terms: D000818:Animals; D000852:Anopheles / Q000276:immunology / Q000469:parasitology; D019138:Antibodies, Blocking / Q000276:immunology; D000953:Antigens, Protozoan / Q000276:immunology; D002455:Cell Division; D003001:Cloning, Molecular; D004306:Dose-Response Relationship, Immunologic; D000939:Epitopes / Q000276:immunology; D015870:Gene Expression; D007074:Immunoglobulin G / Q000032:analysis / Q000276:immunology; D016778:Malaria, Falciparum / Q000235:genetics / Q000276:immunology* / Q000635:transmission; D051379:Mice; D010963:Plasmodium falciparum / Q000235:genetics* / Q000276:immunology*; D011401:Promoter Regions, Genetic; D021382:Protein Sorting Signals / Q000235:genetics; D015800:Protozoan Proteins / Q000235:genetics / Q000276:immunology*; D016052:Protozoan Vaccines / Q000235:genetics / Q000276:immunology; D011993:Recombinant Fusion Proteins / Q000276:immunology* / Q000378:metabolism; D011995:Recombination, Genetic; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000378:metabolism; D006377:T-Lymphocytes, Helper-Inducer / Q000166:cytology / Q000276:immunology; D014611:Vaccination; D014614:Vaccines, Synthetic / Q000276:immunology
chemical_list: D019138:Antibodies, Blocking; D000953:Antigens, Protozoan; D000939:Epitopes; D007074:Immunoglobulin G; C071284:Pfs25 protein, Plasmodium falciparum; D021382:Protein Sorting Signals; D015800:Protozoan Proteins; D016052:Protozoan Vaccines; D011993:Recombinant Fusion Proteins; D014614:Vaccines, Synthetic