Abstract
The lack of suitable animal models for the study of cytoadhesion of P. falciparum-infected erythrocytes (IEs) has necessitated in vitro studies employing a range of cell lines of either human tumour origin (e.g., BeWo and C32 cells) or non-human origin (e.g., CHO cells). Of the human cells available, many were isolated from adults, or derived from a pool of donors (e.g., HBEC-5i). Here we demonstrate, for the first time, the successful isolation of blood outgrowth endothelial cells (BOECs) from frozen stabilates of peripheral blood mononuclear cells obtained from small-volume peripheral blood samples from paediatric malaria patients. BOECs are a sub-population of human endothelial cells, found within the peripheral blood. We demonstrate that these cells express receptors such as Intercellular Adhesion Molecule 1 (ICAM-1/CD54), Endothelial Protein C Receptor (EPCR/CD201), platelet/endothelial cell adhesion molecule 1 (PECAM-1/CD31), Thrombomodulin (CD141), and support adhesion of P. falciparum IEs.
Highlights
The vascular system is one of the largest organs in the human body and is the primary point of interaction between infected erythrocytes (IEs) and the infected host
We used peripheral blood mononuclear cells (PBMCs) freshly isolated from large-volume (30 mL) samples of blood of healthy European adults to optimize the freeze and thaw technique for establishment of blood outgrowth endothelial cells (BOECs) cultures, and for initial phenotypic characterization of the cells (Fig 1A)
The BOECs were analysed via IFA for their surface receptor expression at passage 3, and found to be positive for the adhesion receptors EPCR, ICAM-1, thrombomodulin and PECAM-1 (Fig 1C)
Summary
The vascular system is one of the largest organs in the human body and is the primary point of interaction between infected erythrocytes (IEs) and the infected host. The receptor interactions are mediated by the constituent Duffy binding-like (DBL) and cysteine rich-interdomain (CIDR) domains of the parasite protein family Plasmodium falciparum erythrocyte membrane protein-1 (PfEMP1), encoded by the multi-gene var family (reviewed in[5]). Despite their diversity, the PfEMP1 proteins can be classified into groups A, B and C according to the 5’ un-translated region of the var genes, their genomic location, and their direction of transcription[6,7]. These groups have been associated with clinical presentation of malaria patients, with group A more commonly found among those with severe disease [2,8,9,10,11,12].
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