Abstract
BackgroundTo 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.MethodsA 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.ResultsThe 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.ConclusionsThe 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.
Highlights
To develop a plant-based vaccine against Plasmodium vivax, two P. vivax candidate proteins were chosen
Compared to Plasmodium falciparum vaccine development, less progress has been made on the development of a vivax malarial vaccine due to the lower mortality associated with vivax malaria and limitations in culturing P. vivax in vitro
Based on this work, the MLC chimeric recombinant protein produced in transgenic B. napus plants has potential as a P. vivax vaccine candidate
Summary
To develop a plant-based vaccine against Plasmodium vivax, two P. vivax candidate proteins were chosen. The merozoite surface protein-1 (MSP-1), a major asexual blood stage antigen that is currently considered a strong vaccine candidate. A vaccine to prevent vivax malaria is needed to prevent the morbidity associated with the disease and to prevent the potential spread of malaria due to the reactivation of P. vivax hypnozoites in non-endemic areas. The region containing the tandem repeats was cloned to make a recombinant protein that has subsequently been used for malaria diagnosis in patients, sero-epidemiological investigations, and the production of monoclonal antibodies. This recombinant protein was reported to be a viable vaccine candidate when assessed for immunogenicity [17,18,19]
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