title: Murine immune responses to a Plasmodium vivax-derived chimeric recombinant protein expressed in Brassica napus.
abstract: BACKGROUND
  To develop a plant-based vaccine against Plasmodium vivax, two P. vivax candidate proteins were chosen. First, the merozoite surface protein-1 (MSP-1), a major asexual blood stage antigen that is currently considered a strong vaccine candidate. Second, the circumsporozoite protein (CSP), a component of sporozoites that contains a B-cell epitope.
  
  
  METHODS
  A synthetic chimeric recombinant 516 bp gene encoding containing PvMSP-1, a Pro-Gly linker motif, and PvCSP was synthesized; the gene, named MLC, encoded a total of 172 amino acids. The recombinant gene was modified with regard to codon usage to optimize gene expression in Brassica napus. The Ti plasmid inducible gene transfer system was used for MLC chimeric recombinant gene expression in B. napus. Gene expression was confirmed by polymerase chain reaction (PCR), beta-glucuronidase reporter gene (GUS) assay, and Western blot.
  
  
  RESULTS
  The MLC chimeric recombinant protein expressed in B. napus had a molecular weight of approximately 25 kDa. It exhibited a clinical sensitivity of 84.21% (n=38) and a clinical specificity of 100% (n=24) as assessed by enzyme-linked immunosorbent assay (ELISA). Oral immunization of BALB/c mice with MLC chimeric recombinant protein successfully induced antigen-specific IgG1 production. Additionally, the Th1-related cytokines IL-12 (p40), TNF, and IFN-γ were significantly increased in the spleens of the BALB/c mice.
  
  
  CONCLUSIONS
  The chimeric MLC recombinant protein produced in B. napus has potential as both as an antigen for diagnosis and as a valuable vaccine candidate for oral immunization against vivax malaria.
authors.affiliation: Institute of Global Environment and Department of Biology, Kyung Hee University, Seoul 130-701, Republic of Korea.
authors.forename: Choonghee
authors.initials: C
authors.lastname: Lee
authors.forename: Hyung-Hwan
authors.initials: HH
authors.lastname: Kim
authors.forename: Kyung Mi
authors.initials: KM
authors.lastname: Choi
authors.forename: Kyung Won
authors.initials: KW
authors.lastname: Chung
authors.forename: Yien Kyoung
authors.initials: YK
authors.lastname: Choi
authors.forename: Mi Jung
authors.initials: MJ
authors.lastname: Jang
authors.forename: Tong-Soo
authors.initials: TS
authors.lastname: Kim
authors.forename: Nam-Jun
authors.initials: NJ
authors.lastname: Chung
authors.forename: Ho-Gun
authors.initials: HG
authors.lastname: Rhie
authors.forename: Ho-Sa
authors.initials: HS
authors.lastname: Lee
authors.forename: Youngjoo
authors.initials: Y
authors.lastname: Sohn
authors.forename: Hyuck
authors.initials: H
authors.lastname: Kim
authors.forename: Sung-Jae
authors.initials: SJ
authors.lastname: Lee
authors.forename: Hyeong-Woo
authors.initials: HW
authors.lastname: Lee
mesh_terms: D000284:Administration, Oral; D000818:Animals; D000913:Antibodies, Protozoan / Q000097:blood; D000953:Antigens, Protozoan / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology; D029688:Brassica napus / Q000235:genetics / Q000378:metabolism*; D003062:Codon; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D015870:Gene Expression; D005822:Genetic Vectors; D007074:Immunoglobulin G / Q000097:blood; D017780:Malaria Vaccines / Q000008:administration & dosage / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology*; D008297:Male; D020066:Merozoite Surface Protein 1 / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology*; D051379:Mice; D008807:Mice, Inbred BALB C; D008970:Molecular Weight; D036741:Plant Tumor-Inducing Plasmids; D010966:Plasmodium vivax / Q000235:genetics / Q000276:immunology*; D015800:Protozoan Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology; D012680:Sensitivity and Specificity; D014614:Vaccines, Synthetic / Q000008:administration & dosage / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology
chemical_list: D000913:Antibodies, Protozoan; D000953:Antigens, Protozoan; D003062:Codon; D007074:Immunoglobulin G; D017780:Malaria Vaccines; D020066:Merozoite Surface Protein 1; D015800:Protozoan Proteins; D011994:Recombinant Proteins; D014614:Vaccines, Synthetic; C041595:circumsporozoite protein, Protozoan