Immunogenicity and in vitro protective efficacy of recombinant Mycobacterium bovis bacille Calmette Guerin (rBCG) expressing the 19 kDa merozoite surface protein-1 (MSP-119) antigen of Plasmodium falciparum
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Vaccine development against the blood-stage malaria parasite is aimed at reducing the pathology of the disease. We constructed a recombinant Mycobacterium bovis bacille Calmette Guerin (rBCG) expressing the 19 kDa C-terminus of Plasmodium falciparum merozoite surface protein-1 (MSP-119) to evaluate its protective ability against merozoite invasion of red blood cells in vitro. A mutated version of MSP-119, previously shown to induce the production of inhibitory but not blocking antibodies, was cloned into a suitable shuttle plasmid and transformed into BCG Japan (designated rBCG016). A native version of the molecule was also cloned into BCG (rBCG026). Recombinant BCG expressing the mutated version of MSP-119 (rBCG016) elicited enhanced specific immune response against the epitope in BALB/c mice as compared to rBCG expressing the native version of the epitope (rBCG026). Sera from rBCG016-immunized mice contained significant levels of specific IgG, especially of the IgG2a subclass, against MSP-119 as determined by enzyme-linked immunosorbent assay. The sera was reactive with fixed P. falciparum merozoites as demonstrated by indirect immunofluorescence assay (IFA) and inhibited merozoite invasion of erythrocytes in vitro. Furthermore, lymphocytes from rBCG016-immunized mice demonstrated higher proliferative response against the MSP-119 antigen as compared to those of rBCG026- and BCG-immunized animals. rBCG expressing the mutated version of MSP-119 of P. falciparum induced enhanced humoral and cellular responses against the parasites paving the way for the rational use of rBCG as a blood-stage malaria vaccine candidate.
KeywordsMalaria Bacille Calmette Guerin Preimmune Seron Merozoite Invasion Falciparum Merozoite
We thank Anthony Holder for the gifts of 12.10 and 1E1 mAbs and recombinant MSP-119 protein. We are also thankful to Nor Rain Abdullah for the gift of 3D7 P. falciparum. We also thank Rapeah Suppian, Jamaruddin Mat Asan, Munira Grainger, Irene Ling, and Judith Green for their technical assistance. This work was supported by the Malaysian Institute of Pharmaceuticals and Nutraceuticals Special Initiative Grant [No. 304/PPSK/6150077.I121]. NAN was supported by the Malaysian National Science Fellowship (NSF).
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