A Multistage Formulation Based on Full-Length CSP and AMA-1 Ectodomain ofPlasmodium vivaxInduces High Antibody Titers and T-cells and Partially Protects Mice Challenged with a TransgenicPlasmodium bergheiParasite

Infections withPlasmodium vivaxare predominant in the Americas, representing 75% of malaria cases. Previously perceived as benign, malaria vivax is, in fact, a highly debilitating and economically important disease. Considering the high complexity of the malaria parasite life cycle, it has been hypothesized that an effective vaccine formulation againstPlasmodiumshould contain multiple antigens expressed in different parasite stages. Based on that, we analyzed a recombinantP. vivaxvaccine formulation mixing the apical membrane antigen 1 ectodomain (PvAMA-1) and a full-length circumsporozoite protein (PvCSP-All(FL)) previously studied by our group, which elicits a potent antibody response in mice. Genetically distinct strains of mice (C57BL/6 and BALB/c) were immunized with the proteins, alone or in combination, in the presence of poly(I:C) adjuvant, a TLR3 agonist. In C57BL/6, high-antibody titers were induced against PvAMA-1 and the three PvCSP variants (VK210, VK247, andP. vivax-like). Meanwhile, mixing PvAMA-1 with PvCSP-All(FL)had no impact on total IgG antibody titers, which were long-lasting. Moreover, antibodies from immunized mice recognized VK210 sporozoites and blood-stage parasites by immunofluorescence assay. However, in the BALB/c model, the antibody response against PvCSP-All(FL)was relatively low. PvAMA-1-specific CD3(+)CD4(+)and CD3(+)CD8(+)T-cell responses were observed in C57BL/6 mice, and the cellular response was impaired by PvCSP-All(FL)combination. More relevant, the multistage vaccine formulation provided partial protection in mice challenged with a transgenicPlasmodium bergheisporozoite expressing the homologous PvCSP protein.