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Memórias do Instituto Oswaldo Cruz

Print version ISSN 0074-0276On-line version ISSN 1678-8060

Abstract

FARIAS, Elizangela et al. A simple, ex vivo phagocytosis assay of Plasmodium vivax merozoites by flow cytometry. Mem. Inst. Oswaldo Cruz [online]. 2019, vol.114, e190158.  Epub Oct 07, 2019. ISSN 1678-8060.  https://doi.org/10.1590/0074-02760190158.

As phagocytosis is the first line of defense against malaria, we developed a phagocytosis assay with Plasmodium vivax (P. vivax) merozoites that can be applied to evaluate vaccine candidates. Briefly, after leukocyte removal with loosely packed cellulose powder in a syringe, P. vivax trophozoites matured to the merozoite-rich schizont stages in the presence of the E64 protease inhibitor. The Percoll gradient-enriched schizonts were chemically disrupted to release merozoites that were submitted to merozoite opsonin-dependent phagocytosis in two phagocytic lines with human and mouse antibodies against the N- and C-terminus of P. vivax Merozoite Surface Protein-1 (Nterm-PvMSP1 and MSP119). The resulting assay is simple and efficient for use as a routine phagocytic assay for the evaluation of merozoite stage vaccine candidates.

Keywords : Plasmodium vivax; merozoites; MSP1; opsonising antibodies.

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