relevant assay to study the adhesion of plasmodium falciparum-infected erythrocytes to the placental epithelium相关分析研究疟原虫的附着力falciparum-infected红细胞胎盘上皮.pdfVIP

Relevant Assay to Study the Adhesion of Plasmodium falciparum-Infected Erythrocytes to the Placental Epithelium 1 1 2 3 1 Philippe Boeuf *, Wina Hasang , Eric Hanssen , Jocelyn D. Glazier , Stephen J. Rogerson 1 Department of Medicine (RMH/WH), The University of Melbourne, Royal Melbourne Hospital, Melbourne, Australia, 2 Department of Biochemistry, LaTrobe University, and Electron Microscopy Unit, Bio21 Institute, The University of Melbourne, Melbourne, Australia, 3 Maternal and Fetal Health Research Centre, School of Medicine, Manchester Academic Health Sciences Centre, University of Manchester, St. Mary’s Hospital, Manchester, United Kingdom Abstract In placental malaria, Plasmodium falciparum-infected erythrocytes adhere to the apical plasma membrane of the placental epithelium, triggering an impairment of placental function detrimental to the fetus. The design of anti-adhesion intervention strategies requires a detailed understanding of the mechanisms involved. However, most adhesion assays lack in vivo relevance and are hardly quantitative. Here, we describe a flow cytometry-based adhesion assay that is fully relevant by using apical epithelial plasma membrane vesicles as the adhesion matrix, and being applicable to infected erythrocytes directly isolated from patients. Adhesion is measured both as the percentage of pathogens bound to epithelial membrane vesicles as well as the mean number of vesicles bound per infected erythrocytes. We show that adhesins alternative to those currently identified could be involved. This demonstrates the power of this assay to advance our understanding of epithelial adhesion of infected erythrocytes and in the design of intervention strategies. Citatio