Sequence polymorphism within erythrocyte binding domain of EBA175 in Indian and African field isolates

Abstract

Invasion of erythrocyte by Plasmodium merozoites is mediated by specific molecular interactions between proteins expressed on merozoite surface and the receptors present on erythrocytes. Erythrocyte binding antigen 175 (EBA175) is one such protein that interacts with the sialic acid residues on glycophorin A present on erythrocytes’ surface during invasion. The FII region (PfFII) of EBA175 has been mapped to be critical for binding to erythrocytes. It is reported that antibodies against FII region blocks binding. Polymorphisms in FII region of EBA175 are already reported. The goal of this study was to investigate whether polymorphism in FII region of African P. falciparum field isolates has any effect on erythrocyte binding and also to find whether antibodies raised against FII region from P. falciparum Malayan Camp strain (Camp) can inhibit erythrocyte binding. Genomic DNA of parasites from the blood samples of P. falciparum infected individuals was isolated and PfFII region from these genomic DNA were amplified, cloned and sequenced. Following sequence analysis, we selected three isolates harboring higher PfFII polymorphisms, expressed them on the surface of COS cells as chimeric proteins using secretory signal and transmembrane segments of Herpex simplex virus glycoprotein D (HSVg D) and tested for their erythrocyte binding ability. We further tested the inhibition of erythrocyte binding of these polymorphic FII regions using anti-campPfF2 antibodies. Our results reveal that the polymorphisms in different field isolates included in this study do not have any significant effect on erythrocyte binding and antibodies raised against FII region of camp strain could inhibit erythrocyte binding by all the polymorphic PfFII. This observation strengthens the possibility that PfFII can be a potential candidate vaccine.