Murine immune responses to a Plasmodium vivax-derived chimeric recombinant protein expressed in Brassica napus

作者：Lee Choonghee; Kim Hyung Hwan; Choi Kyung Mi; Chung Kyung Won; Choi Yien Kyoung; Jang Mi Jung; Kim Tong Soo; Chung Nam Jun; Rhie Ho Gun; Lee Ho Sa; Sohn Youngjoo; Kim Hyuck; Lee Sung Jae^*; Lee Hyeong Woo

来源：Malaria Journal, 2011, 10: 106.

DOI：10.1186/1475-2875-10-106

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-gamma 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.

• 出版日期2011-4-29