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A Robust Phagocytosis Assay to Evaluate the Opsonic Activity of Antibodies against Plasmodium falciparum-Infected Erythrocytes.

Abstract
Infection with Plasmodium falciparum parasites causes the majority of malaria-related morbidity and mortality. Constant exposure to the pathogen leads to the acquisition of antibodies and high levels of antibodies have been associated with clinical protection against malaria. A possible protective mechanism is the opsonization of parasites, or malaria-infected erythrocytes (IEs), for phagocytic clearance. Current assays use adherent or chemically differentiated THP-1 cells to evaluate opsonic antibodies in patients' samples, but these assays are often time consuming and damage the effector cells. We have developed a high throughput flow cytometry-based phagocytosis assay using undifferentiated THP-1 cells to quantify the opsonic activity against late stage P. falciparum-IEs. Opsonic antibodies bound to IEs promote their phagocytic uptake through Fcγ receptors found on THP-1 cells. Moreover, undifferentiated THP-1 cells do not express CD36, a surface scavenger receptor that promotes non-opsonic phagocytosis. This technical advance allows quantification of opsonic antibodies and is an important tool for the performance of large, population-based studies of malaria immunity, and to provide a significant increase in the statistical power for such studies.
AuthorsAndrew Teo, Wina Hasang, Philippe Boeuf, Stephen Rogerson
JournalMethods in molecular biology (Clifton, N.J.) (Methods Mol Biol) Vol. 1325 Pg. 145-52 ( 2015) ISSN: 1940-6029 [Electronic] United States
PMID26450386 (Publication Type: Journal Article)

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