The Site of Bluetongue Virus Attachment to Glycophorins from a Number of Animal Erythrocytes

Abstract

Bluetongue virus (BTV) was shown to agglutinate human, ovine and porcine erythrocytes. Removal of neuraminic acid (NA) from erythrocytes by Vibrio cholerae neuraminidase prevented their agglutination. Haemagglutination was also inhibited by N-acetyl neuraminic acid (NANA), N-glycol neuraminic acid (NGNA) and N-acetyl neuramin-lactose. The ability of BTV to agglutinate trypsin-treated human erythrocytes, which lack the amino-terminal domain and the single N-linked oligosaccharide of glycophorin A, suggests that the virus bound to human erythrocytes via NANA-containing, O-linked oligosaccharides. Glycoproteins with NA-containing oligosaccharides of known structure such as mucin, fetuin, alpha 1-acid glycoprotein, ovomucoid and ovine, porcine, human and equine glycophorin were examined for their ability to inhibit BTV-mediated agglutination of human, ovine and porcine erythrocytes. All glycoproteins containing NANA- or NGNA alpha 2-6GalNAc were capable of inhibiting the agglutination of human and porcine erythrocytes. Treatment of human erythrocytes with Newcastle disease virus neuraminidase and of porcine erythrocytes with Clostridium perfringens neuraminidase to cleave preferentially the NANA- and NGNA alpha 2-3Gal linkages respectively, were shown to have little effect on the ability of the erythrocytes to be agglutinated by BTV. The results suggested that BTV binds to NANA- and NGNA alpha 2-6GalNAc residues in the O-linked oligosaccharides of human and porcine glycophorins respectively and indicated the presence of different binding sites on the virus for erythrocytes from other species.