Evaluating specific adhesion of Plasmodium falciparum-infected erythrocytes to immobilised hyaluronic acid with comparison to binding of mammalian cells.
A feature of infection with Plasmodium falciparum is the ability of parasite-infected erythrocytes to adhere to vascular endothelial cells and accumulate in vital organs, associated with severe clinical disease. Hyaluronic acid was recently identified as a receptor for adhesion and has been implicated in mediating the accumulation of parasites in the placenta. Here, we report in vitro assays to measure specific adhesion of infected erythrocytes to hyaluronic acid that is distinct from binding to chondroitin sulphate A, another glycosaminoglycan implicated as a receptor in placental malaria. In this study, specific adhesion of mature stage infected erythrocytes to hyaluronic acid of high purity immobilised on plastic surfaces was abolished by pre-treating hyaluronic acid with a specific hyaluronate lyase from Streptomyces, whereas the same treatment of chondroitin sulphate A had little effect. Adhesion to hyaluronic acid could not be explained by the presence of chondroitin sulphate A or other glycosaminoglycans in the hyaluronic acid preparations. Chinese hamster ovary cells bound in a similar manner in the assays and confirmed that hyaluronic acid was appropriately immobilised for cell adhesion. In contrast to parasites, these cells did not adhere to chondroitin sulphate A. The adsorption of hyaluronic acid onto plastic surfaces was also confirmed by the use of a specific hyaluronic acid-binding protein. Fixing cells with glutaraldehyde at the completion of adhesion assays reduced the number of parasites remaining adherent to hyaluronic acid, but not to chondroitin sulphate A or CD36. These findings have important implications for understanding and evaluating interactions between P. falciparum and hyaluronic acid that may be involved in disease pathogenesis.
Department of Medicine, University of Melbourne, Royal Melbourne Hospital, Melbourne, Vic. 3050, Australia. beeson@unimelb.edu.au
James G
JG
Beeson
Stephen J
SJ
Rogerson
Graham V
GV
Brown
D000818:Animals; D016466:CHO Cells; D002448:Cell Adhesion; D002478:Cells, Cultured; D002809:Chondroitin Sulfates / Q000378:metabolism; D006224:Cricetinae; D004912:Erythrocytes / Q000378:metabolism / Q000469:parasitology*; D006820:Hyaluronic Acid / Q000378:metabolism*; D008322:Mammals / Q000033:anatomy & histology / Q000378:metabolism; D010963:Plasmodium falciparum / Q000378:metabolism / Q000472:pathogenicity*; D011133:Polysaccharide-Lyases / Q000378:metabolism; D013302:Streptomyces / Q000378:metabolism
D006820:Hyaluronic Acid; D002809:Chondroitin Sulfates; D011133:Polysaccharide-Lyases; C100002:hyaluronate lyase