Please use this identifier to cite or link to this item:
https://hdl.handle.net/2440/83596
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Type: | Journal article |
Title: | Efficient measurement of opsonising antibodies to plasmodium falciparum merozoites |
Author: | Hill, D. Eriksson, E. Carmagnac, A. Wilson, D. Cowman, A. Hansen, D. Schofield, L. |
Citation: | PLoS One, 2012; 7(12):e15692-1-e15692-10 |
Publisher: | Public Library of Science |
Issue Date: | 2012 |
ISSN: | 1932-6203 1932-6203 |
Editor: | Braga, E.M. |
Statement of Responsibility: | Danika L. Hill, Emily M. Eriksson, Amandine B. Carmagnac, Danny W. Wilson, Alan F. Cowman, Diana S. Hansen, Louis Schofield |
Abstract: | BACKGROUND Antibodies targeting merozoites are important in protection from malaria. Therefore, merozoite surface proteins are attractive vaccine candidates. There is a need for robust functional assays to investigate mechanisms of acquired immunity and vaccine efficacy. To date, the study of merozoite phagocytosis has been confounded by the complexity and variability of in vitro assays. METHODOLOGY/PRINCIPAL FINDINGS We have developed a new flow cytometry-based merozoite phagocytosis assay. An optimized merozoite preparation technique produced high yields of merozoites separated from haemozoin. Phagocytosis by the undifferentiated THP-1 monocytic cell line was mediated only by Fc Receptors, and was therefore ideal for studying opsonising antibody responses. The assay showed robust phagocytosis with highly diluted immune sera and strong inter-assay correlation. The assay effectively measured differences in opsonisation-dependent phagocytosis among individuals. CONCLUSIONS/SIGNIFICANCE This highly reproducible assay has potential applications in assessing the role of opsonic phagocytosis in naturally acquired immunity and vaccine trials. |
Keywords: | Monocytes Cells, Cultured Humans Plasmodium falciparum Malaria, Falciparum Receptors, Fc Antibodies, Protozoan Antigens, Protozoan Fluorescent Antibody Technique, Indirect Immunoblotting Flow Cytometry Reproducibility of Results Phagocytosis Adolescent Adult Middle Aged Child Child, Preschool Opsonin Proteins Merozoites Young Adult |
Rights: | © 2012 Hill et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. |
DOI: | 10.1371/journal.pone.0051692 |
Grant ID: | http://purl.org/au-research/grants/nhmrc/1035715 http://purl.org/au-research/grants/nhmrc/1031212 |
Published version: | http://dx.doi.org/10.1371/journal.pone.0051692 |
Appears in Collections: | Aurora harvest Molecular and Biomedical Science publications |
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hdl_83596.pdf | Published version | 934.45 kB | Adobe PDF | View/Open |
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