A method for positive and negative selection of Plasmodium falciparum platelet-mediated clumping parasites and investigation of the role of CD36.
Platelet-mediated clumping of Plasmodium falciparum infected erythrocytes (IEs) is a frequently observed parasite adhesion phenotype. The importance of clumping in severe malaria and the molecular mechanisms behind this phenomenon are incompletely understood. Three platelet surface molecules have pr...
Saved in:
| Main Authors: | , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Public Library of Science (PLoS)
2013-01-01
|
| Series: | PLoS ONE |
| Online Access: | https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0055453&type=printable |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1850137424307421184 |
|---|---|
| author | Mònica Arman Yvonne Adams Gabriella Lindergard J Alexandra Rowe |
| author_facet | Mònica Arman Yvonne Adams Gabriella Lindergard J Alexandra Rowe |
| author_sort | Mònica Arman |
| collection | DOAJ |
| description | Platelet-mediated clumping of Plasmodium falciparum infected erythrocytes (IEs) is a frequently observed parasite adhesion phenotype. The importance of clumping in severe malaria and the molecular mechanisms behind this phenomenon are incompletely understood. Three platelet surface molecules have previously been identified as clumping receptors: CD36, globular C1q receptor (gC1qR/HABP1/p32), and P-selectin (CD62P), but the parasite ligands mediating this phenotype are unknown. The aim of this work was to develop a selection method to facilitate investigations of the molecular mechanisms of clumping in selected P. falciparum lines. Magnetic beads coated with anti-platelet antibodies were used to positively and negatively select clumping IEs from P. falciparum strains IT, HB3, 3D7 and Dd2. Clumping in all four positively selected parasite lines was abolished by antibodies to CD36, but was not affected by antibodies to gC1qR or P-selectin. Clumping positive lines showed significantly higher binding to CD36 than clumping negative lines in flow adhesion assays (strains IT, HB3 and 3D7, p<0.05 for all strains, paired t test) and static assays (strain Dd2, p<0.0001 paired t test). However, clumping negative lines IT, HB3 and 3D7 did show some binding to CD36 under flow conditions, indicating that CD36-binding is not sufficient for clumping. These data show that CD36-dependent clumping positive and negative lines can easily be selected from P. falciparum laboratory strains. CD36-binding is necessary but not sufficient for clumping, and the molecular differences between clumping positive and negative parasite lines responsible for the phenotype require further investigation. |
| format | Article |
| id | doaj-art-c310b71696eb4ae8bc46e29d63fa3e3b |
| institution | OA Journals |
| issn | 1932-6203 |
| language | English |
| publishDate | 2013-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj-art-c310b71696eb4ae8bc46e29d63fa3e3b2025-08-20T02:30:50ZengPublic Library of Science (PLoS)PLoS ONE1932-62032013-01-0182e5545310.1371/journal.pone.0055453A method for positive and negative selection of Plasmodium falciparum platelet-mediated clumping parasites and investigation of the role of CD36.Mònica ArmanYvonne AdamsGabriella LindergardJ Alexandra RowePlatelet-mediated clumping of Plasmodium falciparum infected erythrocytes (IEs) is a frequently observed parasite adhesion phenotype. The importance of clumping in severe malaria and the molecular mechanisms behind this phenomenon are incompletely understood. Three platelet surface molecules have previously been identified as clumping receptors: CD36, globular C1q receptor (gC1qR/HABP1/p32), and P-selectin (CD62P), but the parasite ligands mediating this phenotype are unknown. The aim of this work was to develop a selection method to facilitate investigations of the molecular mechanisms of clumping in selected P. falciparum lines. Magnetic beads coated with anti-platelet antibodies were used to positively and negatively select clumping IEs from P. falciparum strains IT, HB3, 3D7 and Dd2. Clumping in all four positively selected parasite lines was abolished by antibodies to CD36, but was not affected by antibodies to gC1qR or P-selectin. Clumping positive lines showed significantly higher binding to CD36 than clumping negative lines in flow adhesion assays (strains IT, HB3 and 3D7, p<0.05 for all strains, paired t test) and static assays (strain Dd2, p<0.0001 paired t test). However, clumping negative lines IT, HB3 and 3D7 did show some binding to CD36 under flow conditions, indicating that CD36-binding is not sufficient for clumping. These data show that CD36-dependent clumping positive and negative lines can easily be selected from P. falciparum laboratory strains. CD36-binding is necessary but not sufficient for clumping, and the molecular differences between clumping positive and negative parasite lines responsible for the phenotype require further investigation.https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0055453&type=printable |
| spellingShingle | Mònica Arman Yvonne Adams Gabriella Lindergard J Alexandra Rowe A method for positive and negative selection of Plasmodium falciparum platelet-mediated clumping parasites and investigation of the role of CD36. PLoS ONE |
| title | A method for positive and negative selection of Plasmodium falciparum platelet-mediated clumping parasites and investigation of the role of CD36. |
| title_full | A method for positive and negative selection of Plasmodium falciparum platelet-mediated clumping parasites and investigation of the role of CD36. |
| title_fullStr | A method for positive and negative selection of Plasmodium falciparum platelet-mediated clumping parasites and investigation of the role of CD36. |
| title_full_unstemmed | A method for positive and negative selection of Plasmodium falciparum platelet-mediated clumping parasites and investigation of the role of CD36. |
| title_short | A method for positive and negative selection of Plasmodium falciparum platelet-mediated clumping parasites and investigation of the role of CD36. |
| title_sort | method for positive and negative selection of plasmodium falciparum platelet mediated clumping parasites and investigation of the role of cd36 |
| url | https://journals.plos.org/plosone/article/file?id=10.1371/journal.pone.0055453&type=printable |
| work_keys_str_mv | AT monicaarman amethodforpositiveandnegativeselectionofplasmodiumfalciparumplateletmediatedclumpingparasitesandinvestigationoftheroleofcd36 AT yvonneadams amethodforpositiveandnegativeselectionofplasmodiumfalciparumplateletmediatedclumpingparasitesandinvestigationoftheroleofcd36 AT gabriellalindergard amethodforpositiveandnegativeselectionofplasmodiumfalciparumplateletmediatedclumpingparasitesandinvestigationoftheroleofcd36 AT jalexandrarowe amethodforpositiveandnegativeselectionofplasmodiumfalciparumplateletmediatedclumpingparasitesandinvestigationoftheroleofcd36 AT monicaarman methodforpositiveandnegativeselectionofplasmodiumfalciparumplateletmediatedclumpingparasitesandinvestigationoftheroleofcd36 AT yvonneadams methodforpositiveandnegativeselectionofplasmodiumfalciparumplateletmediatedclumpingparasitesandinvestigationoftheroleofcd36 AT gabriellalindergard methodforpositiveandnegativeselectionofplasmodiumfalciparumplateletmediatedclumpingparasitesandinvestigationoftheroleofcd36 AT jalexandrarowe methodforpositiveandnegativeselectionofplasmodiumfalciparumplateletmediatedclumpingparasitesandinvestigationoftheroleofcd36 |