High Yield Purification of Plasmodium falciparum Merozoites For Use in Opsonizing Antibody Assays
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 challeng...
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| Format: | Online |
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MyJove Corporation
2014
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| Online Access: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4217647/ |
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pubmed-4217647 |
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pubmed-42176472014-11-13 High Yield Purification of Plasmodium falciparum Merozoites For Use in Opsonizing Antibody Assays Hill, Danika L. Eriksson, Emily M. Schofield, Louis Immunology 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. MyJove Corporation 2014-07-17 /pmc/articles/PMC4217647/ /pubmed/25078358 http://dx.doi.org/10.3791/51590 Text en Copyright © 2014, Journal of Visualized Experiments http://creativecommons.org/licenses/by-nc-nd/3.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs 3.0 Unported License. To view a copy of this license, visithttp://creativecommons.org/licenses/by-nc-nd/3.0/ |
| repository_type |
Open Access Journal |
| institution_category |
Foreign Institution |
| institution |
US National Center for Biotechnology Information |
| building |
NCBI PubMed |
| collection |
Online Access |
| language |
English |
| format |
Online |
| author |
Hill, Danika L. Eriksson, Emily M. Schofield, Louis |
| spellingShingle |
Hill, Danika L. Eriksson, Emily M. Schofield, Louis High Yield Purification of Plasmodium falciparum Merozoites For Use in Opsonizing Antibody Assays |
| author_facet |
Hill, Danika L. Eriksson, Emily M. Schofield, Louis |
| author_sort |
Hill, Danika L. |
| title |
High Yield Purification of Plasmodium falciparum Merozoites For Use in Opsonizing Antibody Assays |
| title_short |
High Yield Purification of Plasmodium falciparum Merozoites For Use in Opsonizing Antibody Assays |
| title_full |
High Yield Purification of Plasmodium falciparum Merozoites For Use in Opsonizing Antibody Assays |
| title_fullStr |
High Yield Purification of Plasmodium falciparum Merozoites For Use in Opsonizing Antibody Assays |
| title_full_unstemmed |
High Yield Purification of Plasmodium falciparum Merozoites For Use in Opsonizing Antibody Assays |
| title_sort |
high yield purification of plasmodium falciparum merozoites for use in opsonizing antibody assays |
| description |
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. |
| publisher |
MyJove Corporation |
| publishDate |
2014 |
| url |
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4217647/ |
| _version_ |
1613151577191743488 |