A Bicistronic DNA Vaccine Containing Apical Membrane Antigen 1 and Merozoite Surface Protein 4/5 Can Prime Humoral and Cellular Immune Responses and Partially Protect Mice against Virulent Plasmodium chabaudi adami DS Malaria

Department of Biochemistry and Molecular Biology, Monash University, Clayton, Australia.
Infection and Immunity (Impact Factor: 3.73). 11/2004; 72(10):5565-73. DOI: 10.1128/IAI.72.10.5565-5573.2004
Source: PubMed


The ultimate malaria vaccine will require the delivery of multiple antigens from different stages of the complex malaria life cycle. In order to efficiently deliver multiple antigens with use of DNA vaccine technology, new antigen delivery systems must be assessed. This study utilized a bicistronic vector construct, containing an internal ribosome entry site, expressing a combination of malarial candidate antigens: merozoite surface protein 4/5 (MSP4/5) (fused to a monocyte chemotactic protein 3 chemoattractant sequence) and apical membrane antigen 1 (AMA-1) (fused to a tissue plasminogen activator secretion signal). Transfection of COS 7 cells with bicistronic plasmids resulted in production and secretion of both AMA-1 and MSP4/5 in vitro. Vaccination of BALB/c mice via intraepidermal gene gun and intramuscular routes against AMA-1 and MSP4/5 resulted in antibody production and significant in vitro proliferation of splenocytes stimulated by both AMA-1 and MSP4/5. Survival of BALB/c mice vaccinated with bicistronic constructs after lethal Plasmodium chabaudi adami DS erythrocytic-stage challenge was variable, although significant increases in survival and reductions in peak parasitemia were observed in several challenge trials when the vaccine was delivered by the intramuscular route. This study using a murine model demonstrates that the delivery of malarial antigens via bicistronic vectors is feasible. Further experimentation with bicistronic delivery systems is required for the optimization and refinement of DNA vaccines to effectively prime protective immune responses against malaria.

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    • "Also, previous studies in IFN-g knockout mice have shown that the absence of IFN-g can increase the recovery of infective larvae in mice, suggesting its role in parasite killing (Babu et al., 2000). The use of multiple antigens in a cocktail mode for a complex multicellular parasite would be necessary to achieve a high level of protection (Rainczuk et al., 2004; Mendez et al., 2005). A recent study by Anand et al. (2011) showed that an abundant larval transcript (ALT) and venom allergen homologue (VAH) cocktail vaccine conferred significantly high protection. "
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    • "reported as one of the IFN-γ-inducing antigens of MAP, also strongly induced IL-10 from macrophages obtained from infected calves [14]. Bicistronic vectors have been used to design DNA vaccine against HIV infection, which contained gp120 and GM-CSF gene [17], bicistronic DNA vaccine containing apical membrane antigen 1 and merozoite surface protein 4/5 can prime humoral and cellular immune responses and partially protect mice against virulent plasmodium chabaudi adami DS malaria [18], and a bicistronic woodchuck hepatitis virus core and gamma interferon DNA vaccine can protect from hepatitis [19]. Recently from our laboratory, Kadam et al. [20], have reported that coexpression of IFNγ with a 16.8 kDa gene of MAP can enhance immunogenicity of DNA vaccine using the same protein. "
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    • "First bleed was done day 0 after the second immunization while the second bleed was done 14 days after the last boost. In each case pooled sera were obtained and the antibody response was measured by ELISA as previously described [33] against crude parasite antigen, 18 ␮g/ml. Parasite crude antigen at concentration of 18 ␮g/ml, in coating buffer (pH 9.6), was used and 100 ␮l was added to each well of the ELISA plates (Nunc, Copenhagen, Denmark) and incubated at 4 • C overnight. "
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