Mem Inst Oswaldo Cruz, Rio de Janeiro, VOLUME 114 | SEPTEMBER 2019
Short communication

A simple, ex vivo phagocytosis assay of Plasmodium vivax merozoites by flow cytometry

Elizangela Farias1,2, Fhabiane Bezerra1, Djane Clarys Baia-da-Silva3,Yury Chaves2, Tatiana Bacry Cardoza2, Maria Edilene Almeida2, Lucas Barbosa Oliveira2, Pritesh Lalwani1,2, Patrícia P Orlandi2, Marcus Vinicius Guimaraes Lacerda2,3, Stefanie Lopes2, Paulo Afonso Nogueira+,2

1Universidade Federal do Amazonas, Manaus, Amazonas, Brazil
2Instituto Leônidas e Maria Deane (Fiocruz Amazônia), Manaus, Amazonas, Brazil
3Fundação de Medicina Tropical Dr. Heitor Vieira Dourado, Manaus, Amazonas, Brazil

DOI: 10.1590/0074-02760190158
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ABSTRACT

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-terminal domains of P. vivax Merozoite Surface Protein-1. The resulting assay is simple and efficient for use as a routine phagocytic assay for the evaluation of merozoite stage vaccine candidates.

Correspondent author: paulo.nogueira@fiocruz.br
https://orcid.org/0000-0001-9677-7225
Received 30 April 2019
Accepted 04 September 2019

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