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Clinical Immunology | Spotlight

An Opsonic Phagocytosis Assay for Plasmodium falciparum Sporozoites

Ryan W. J. Steel, Brandon K. Sack, Moriya Tsuji, Mary Jane L. Navarro, Will Betz, Matt E. Fishbaugher, Erika L. Flannery, Stefan H. I. Kappe
Patricia P. Wilkins, Editor
Ryan W. J. Steel
aCenter for Infectious Disease Research, Seattle, Washington, USA
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Brandon K. Sack
aCenter for Infectious Disease Research, Seattle, Washington, USA
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Moriya Tsuji
bHIV and Malaria Vaccine Program, Aaron Diamond AIDS Research Center, New York, New York, USA
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Mary Jane L. Navarro
aCenter for Infectious Disease Research, Seattle, Washington, USA
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Will Betz
aCenter for Infectious Disease Research, Seattle, Washington, USA
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Matt E. Fishbaugher
aCenter for Infectious Disease Research, Seattle, Washington, USA
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Erika L. Flannery
aCenter for Infectious Disease Research, Seattle, Washington, USA
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Stefan H. I. Kappe
aCenter for Infectious Disease Research, Seattle, Washington, USA
cDepartment of Global Health, University of Washington, Seattle, Washington, USA
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Patricia P. Wilkins
CDC
Roles: Editor
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DOI: 10.1128/CVI.00445-16
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ABSTRACT

Plasmodium falciparum malaria remains the deadliest parasitic disease worldwide. Vaccines targeting the preerythrocytic sporozoite and liver stages have the potential to entirely prevent blood-stage infection and disease, as well as onward transmission. Sporozoite surface and secreted proteins are leading candidates for inclusion in a preerythrocytic stage-specific, antibody-based vaccine. Preclinical functional assays to identify humoral correlates of protection in vitro and to validate novel sporozoite protein targets for inclusion in multisubunit vaccines currently do not consider the interaction of sporozoite-targeting antibodies with other components of the immune system. Here, we describe the development of a simple flow cytometric assay to quantitatively assess the ability of antibodies directed against P. falciparum sporozoites to facilitate their phagocytosis. We demonstrate that this sporozoite opsonic phagocytosis assay (SOPA) is compatible with both monoclonal antibodies and human immune serum and can be performed using cryopreserved P. falciparum sporozoites. This simple, accessible assay will aid with the assessment of antibody responses to vaccination with Plasmodium antigens and their interaction with phagocytic cells of the immune system.

FOOTNOTES

    • Received 12 September 2016.
    • Returned for modification 2 October 2016.
    • Accepted 8 November 2016.
    • Accepted manuscript posted online 23 November 2016.
  • Supplemental material for this article may be found at https://doi.org/10.1128/CVI.00445-16 .

  • Copyright © 2017 American Society for Microbiology.

All Rights Reserved .

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An Opsonic Phagocytosis Assay for Plasmodium falciparum Sporozoites
Ryan W. J. Steel, Brandon K. Sack, Moriya Tsuji, Mary Jane L. Navarro, Will Betz, Matt E. Fishbaugher, Erika L. Flannery, Stefan H. I. Kappe
Clinical and Vaccine Immunology Feb 2017, 24 (2) e00445-16; DOI: 10.1128/CVI.00445-16

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An Opsonic Phagocytosis Assay for Plasmodium falciparum Sporozoites
Ryan W. J. Steel, Brandon K. Sack, Moriya Tsuji, Mary Jane L. Navarro, Will Betz, Matt E. Fishbaugher, Erika L. Flannery, Stefan H. I. Kappe
Clinical and Vaccine Immunology Feb 2017, 24 (2) e00445-16; DOI: 10.1128/CVI.00445-16
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KEYWORDS

Antibodies, Protozoan
Immunoassay
Opsonin Proteins
phagocytosis
Plasmodium falciparum
Sporozoites
Plasmodium falciparum
antibodies
assay development
malaria
opsonization
phagocytosis
sporozoite
vaccines

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