A whole blood assay for antibody dependent phagocytosis of Plasmodium falciparum infected erythrocytes
Abstract Background Antibodies are used to protect against Plasmodium falciparum malaria. One antibody target, the variant surface antigens, is expressed on infected erythrocytes (IEs). Antibodies to these antigens can either block IE sequestration in the tissues, facilitate natural killer cell-medi...
Saved in:
| Main Authors: | , , , , , , , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Nature Portfolio
2025-07-01
|
| Series: | Communications Medicine |
| Online Access: | https://doi.org/10.1038/s43856-025-00989-2 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1849332192339034112 |
|---|---|
| author | Dilini Rathnayake Wina Hasang Alexander Macpherson HongHua Ding Laurens Manning Moses Laman Maria Ome-Kaius Holger W. Unger Feiko Ter Kuile Mwayi Madanitsa Bruce Wines P. Mark Hogarth Elizabeth H. Aitken Stephen J. Rogerson |
| author_facet | Dilini Rathnayake Wina Hasang Alexander Macpherson HongHua Ding Laurens Manning Moses Laman Maria Ome-Kaius Holger W. Unger Feiko Ter Kuile Mwayi Madanitsa Bruce Wines P. Mark Hogarth Elizabeth H. Aitken Stephen J. Rogerson |
| author_sort | Dilini Rathnayake |
| collection | DOAJ |
| description | Abstract Background Antibodies are used to protect against Plasmodium falciparum malaria. One antibody target, the variant surface antigens, is expressed on infected erythrocytes (IEs). Antibodies to these antigens can either block IE sequestration in the tissues, facilitate natural killer cell-mediated killing, or opsonise IEs for phagocytic clearance by neutrophils and monocytes. Methods We developed a high-throughput assay to measure antibody-dependent neutrophil phagocytosis (ADNP) and antibody-dependent cellular phagocytosis (ADCP, by blood monocytes) in the same sample of fresh whole blood. Results Here we show that immune plasma mediates ADNP and ADCP in a concentration-dependent manner. Uptake is greater in the presence of complement proteins and is largely dependent on the expression of P. falciparum Erythrocyte Membrane Protein 1 located on the IE surface. Plasma from pregnant Papua New Guinean women with and without placental malaria shows that ADNP and ADCP are associated with protection from placental malaria. ADNP, but not ADCP, using IEs expressing IT4VAR19 (a PfEMP1 variant that binds to endothelial protein C receptor through a DC8 domain cassette) is higher at hospital presentation in children with uncomplicated malaria than in severe malaria. In pregnant women, ADNP and ADCP in whole blood are strongly correlated with one another (Spearman’s rho = 0.90), but not with ADNP or ADCP using purified neutrophils and monocytes in the absence of complement proteins. Conclusions The whole blood assay is a powerful new tool to assess functional antibodies that may protect against P. falciparum malaria. It allows simultaneous measurement of phagocytosis of opsonised IEs by monocytes and neutrophils. |
| format | Article |
| id | doaj-art-e2d37fa8fe4c4da3bbf27d22b8ae0e31 |
| institution | Kabale University |
| issn | 2730-664X |
| language | English |
| publishDate | 2025-07-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Communications Medicine |
| spelling | doaj-art-e2d37fa8fe4c4da3bbf27d22b8ae0e312025-08-20T03:46:16ZengNature PortfolioCommunications Medicine2730-664X2025-07-015111210.1038/s43856-025-00989-2A whole blood assay for antibody dependent phagocytosis of Plasmodium falciparum infected erythrocytesDilini Rathnayake0Wina Hasang1Alexander Macpherson2HongHua Ding3Laurens Manning4Moses Laman5Maria Ome-Kaius6Holger W. Unger7Feiko Ter Kuile8Mwayi Madanitsa9Bruce Wines10P. Mark Hogarth11Elizabeth H. Aitken12Stephen J. Rogerson13Department of Medicine (RMH), Peter Doherty Institute of Infection and Immunity, University of MelbourneDepartment of Infectious Diseases, Peter Doherty Institute of Infection and Immunity, University of MelbourneDiscovery Chemistry Research and Technologies, Eli Lilly & Co.Department of Infectious Diseases, Peter Doherty Institute of Infection and Immunity, University of MelbourneUWA Medical School, The University of Western AustraliaPapua New Guinea. Institute of Medical ResearchPapua New Guinea. Institute of Medical ResearchGlobal and Tropical Health Division, Menzies School of Health Research, Charles Darwin UniversityDepartment of Clinical Sciences, Liverpool School of Tropical MedicineDepartment of Clinical Sciences, Academy of Medical Sciences, Malawi University of Science and TechnologyImmune Therapies Group, Centre for Biomedical Research, Burnet InstituteImmune Therapies Group, Centre for Biomedical Research, Burnet InstituteDepartment of Infectious Diseases, Peter Doherty Institute of Infection and Immunity, University of MelbourneDepartment of Medicine (RMH), Peter Doherty Institute of Infection and Immunity, University of MelbourneAbstract Background Antibodies are used to protect against Plasmodium falciparum malaria. One antibody target, the variant surface antigens, is expressed on infected erythrocytes (IEs). Antibodies to these antigens can either block IE sequestration in the tissues, facilitate natural killer cell-mediated killing, or opsonise IEs for phagocytic clearance by neutrophils and monocytes. Methods We developed a high-throughput assay to measure antibody-dependent neutrophil phagocytosis (ADNP) and antibody-dependent cellular phagocytosis (ADCP, by blood monocytes) in the same sample of fresh whole blood. Results Here we show that immune plasma mediates ADNP and ADCP in a concentration-dependent manner. Uptake is greater in the presence of complement proteins and is largely dependent on the expression of P. falciparum Erythrocyte Membrane Protein 1 located on the IE surface. Plasma from pregnant Papua New Guinean women with and without placental malaria shows that ADNP and ADCP are associated with protection from placental malaria. ADNP, but not ADCP, using IEs expressing IT4VAR19 (a PfEMP1 variant that binds to endothelial protein C receptor through a DC8 domain cassette) is higher at hospital presentation in children with uncomplicated malaria than in severe malaria. In pregnant women, ADNP and ADCP in whole blood are strongly correlated with one another (Spearman’s rho = 0.90), but not with ADNP or ADCP using purified neutrophils and monocytes in the absence of complement proteins. Conclusions The whole blood assay is a powerful new tool to assess functional antibodies that may protect against P. falciparum malaria. It allows simultaneous measurement of phagocytosis of opsonised IEs by monocytes and neutrophils.https://doi.org/10.1038/s43856-025-00989-2 |
| spellingShingle | Dilini Rathnayake Wina Hasang Alexander Macpherson HongHua Ding Laurens Manning Moses Laman Maria Ome-Kaius Holger W. Unger Feiko Ter Kuile Mwayi Madanitsa Bruce Wines P. Mark Hogarth Elizabeth H. Aitken Stephen J. Rogerson A whole blood assay for antibody dependent phagocytosis of Plasmodium falciparum infected erythrocytes Communications Medicine |
| title | A whole blood assay for antibody dependent phagocytosis of Plasmodium falciparum infected erythrocytes |
| title_full | A whole blood assay for antibody dependent phagocytosis of Plasmodium falciparum infected erythrocytes |
| title_fullStr | A whole blood assay for antibody dependent phagocytosis of Plasmodium falciparum infected erythrocytes |
| title_full_unstemmed | A whole blood assay for antibody dependent phagocytosis of Plasmodium falciparum infected erythrocytes |
| title_short | A whole blood assay for antibody dependent phagocytosis of Plasmodium falciparum infected erythrocytes |
| title_sort | whole blood assay for antibody dependent phagocytosis of plasmodium falciparum infected erythrocytes |
| url | https://doi.org/10.1038/s43856-025-00989-2 |
| work_keys_str_mv | AT dilinirathnayake awholebloodassayforantibodydependentphagocytosisofplasmodiumfalciparuminfectederythrocytes AT winahasang awholebloodassayforantibodydependentphagocytosisofplasmodiumfalciparuminfectederythrocytes AT alexandermacpherson awholebloodassayforantibodydependentphagocytosisofplasmodiumfalciparuminfectederythrocytes AT honghuading awholebloodassayforantibodydependentphagocytosisofplasmodiumfalciparuminfectederythrocytes AT laurensmanning awholebloodassayforantibodydependentphagocytosisofplasmodiumfalciparuminfectederythrocytes AT moseslaman awholebloodassayforantibodydependentphagocytosisofplasmodiumfalciparuminfectederythrocytes AT mariaomekaius awholebloodassayforantibodydependentphagocytosisofplasmodiumfalciparuminfectederythrocytes AT holgerwunger awholebloodassayforantibodydependentphagocytosisofplasmodiumfalciparuminfectederythrocytes AT feikoterkuile awholebloodassayforantibodydependentphagocytosisofplasmodiumfalciparuminfectederythrocytes AT mwayimadanitsa awholebloodassayforantibodydependentphagocytosisofplasmodiumfalciparuminfectederythrocytes AT brucewines awholebloodassayforantibodydependentphagocytosisofplasmodiumfalciparuminfectederythrocytes AT pmarkhogarth awholebloodassayforantibodydependentphagocytosisofplasmodiumfalciparuminfectederythrocytes AT elizabethhaitken awholebloodassayforantibodydependentphagocytosisofplasmodiumfalciparuminfectederythrocytes AT stephenjrogerson awholebloodassayforantibodydependentphagocytosisofplasmodiumfalciparuminfectederythrocytes AT dilinirathnayake wholebloodassayforantibodydependentphagocytosisofplasmodiumfalciparuminfectederythrocytes AT winahasang wholebloodassayforantibodydependentphagocytosisofplasmodiumfalciparuminfectederythrocytes AT alexandermacpherson wholebloodassayforantibodydependentphagocytosisofplasmodiumfalciparuminfectederythrocytes AT honghuading wholebloodassayforantibodydependentphagocytosisofplasmodiumfalciparuminfectederythrocytes AT laurensmanning wholebloodassayforantibodydependentphagocytosisofplasmodiumfalciparuminfectederythrocytes AT moseslaman wholebloodassayforantibodydependentphagocytosisofplasmodiumfalciparuminfectederythrocytes AT mariaomekaius wholebloodassayforantibodydependentphagocytosisofplasmodiumfalciparuminfectederythrocytes AT holgerwunger wholebloodassayforantibodydependentphagocytosisofplasmodiumfalciparuminfectederythrocytes AT feikoterkuile wholebloodassayforantibodydependentphagocytosisofplasmodiumfalciparuminfectederythrocytes AT mwayimadanitsa wholebloodassayforantibodydependentphagocytosisofplasmodiumfalciparuminfectederythrocytes AT brucewines wholebloodassayforantibodydependentphagocytosisofplasmodiumfalciparuminfectederythrocytes AT pmarkhogarth wholebloodassayforantibodydependentphagocytosisofplasmodiumfalciparuminfectederythrocytes AT elizabethhaitken wholebloodassayforantibodydependentphagocytosisofplasmodiumfalciparuminfectederythrocytes AT stephenjrogerson wholebloodassayforantibodydependentphagocytosisofplasmodiumfalciparuminfectederythrocytes |