High Yield Purification of Plasmodium Falciparum Merozoites for Use in Opsonizing Antibody Assays

Overview

Journal J Vis Exp

Specialties Biology
General Medicine

Date 2014 Aug 1

PMID 25078358

Citations 10

Authors

Danika L Hill

Emily M Eriksson

Louis Schofield

Affiliations

Soon will be listed here.

Abstract

Plasmodium falciparum merozoite antigens are under development as potential malaria vaccines. One aspect of immunity against malaria is the removal of free merozoites from the blood by phagocytic cells. However assessing the functional efficacy of merozoite specific opsonizing antibodies is challenging due to the short half-life of merozoites and the variability of primary phagocytic cells. Described in detail herein is a method for generating viable merozoites using the E64 protease inhibitor, and an assay of merozoite opsonin-dependent phagocytosis using the pro-monocytic cell line THP-1. E64 prevents schizont rupture while allowing the development of merozoites which are released by filtration of treated schizonts. Ethidium bromide labelled merozoites are opsonized with human plasma samples and added to THP-1 cells. Phagocytosis is assessed by a standardized high throughput protocol. Viable merozoites are a valuable resource for assessing numerous aspects of P. falciparum biology, including assessment of immune function. Antibody levels measured by this assay are associated with clinical immunity to malaria in naturally exposed individuals. The assay may also be of use for assessing vaccine induced antibodies.

Citing Articles

Merozoites Obtained through a Simpler Method Do Not Survive in Classically Activated Macrophages.

Rodrigues P, Azeredo M, Almeida N, de Almeida G, Wanderley J, Seabra S Microorganisms. 2024; 12(1).

PMID: 38257932 PMC: 10818340. DOI: 10.3390/microorganisms12010105.

Cytolytic circumsporozoite-specific memory CD4 T cell clones are expanded during Plasmodium falciparum infection.

Furtado R, Paul M, Zhang J, Sung J, Karell P, Kim R Nat Commun. 2023; 14(1):7726.

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Susceptibility to malaria in fulani, Bariba, Otamari and gando individuals living in sympatry in Benin: Role of opsonizing antibodies to Plasmodium falciparum merozoites.

Fall A, Kana I, Garcia-Senosiain A, Henry B, Dechavanne C, Garcia A Heliyon. 2023; 9(1):e13092.

PMID: 36711279 PMC: 9879790. DOI: 10.1016/j.heliyon.2023.e13092.

Naturally acquired antibodies from Beninese infants promote Plasmodium falciparum merozoite-phagocytosis by human blood leukocytes: implications for control of asymptomatic malaria infections.

Fall A, Kana I, Dechavanne C, Garcia-Senosiain A, Guitard E, Milet J Malar J. 2022; 21(1):356.

PMID: 36447200 PMC: 9707106. DOI: 10.1186/s12936-022-04361-w.

Treatment of Plasmodium falciparum merozoites with the protease inhibitor E64 and mechanical filtration increases their susceptibility to complement activation.

Stoute J, Landmesser M, Biryukov S PLoS One. 2020; 15(8):e0237786.

PMID: 32822376 PMC: 7442247. DOI: 10.1371/journal.pone.0237786.

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