Pig Paramyxovirus of the Blue Eye Disease Binding to a 116 kDa Glycoprotein Expressed in Pig Neuronal Membranes

Abstract

Pig paramyxovirus of the blue eye disease (PPBED) is a novel member of the paramyxoviridae family which infects pigs. In neonatal pigs it causes neurological damage, whereas in adult pigs it affects the reproductive function. As PPBED damages the new‐born pig central nervous system (CNS), it is important to study whether PPBED binds to the membrane proteins of all brain tissue, or selectively binds to neuronal tissue of the brain stem, olfactory bulb, hippocampus, cerebellum, frontal, temporal and parietal brain cortex. It is also important to establish whether it also infects neurones obtained from new‐born, 60‐day‐old and adult pigs, and the role of carbohydrate residues in virus binding. The effect on virus binding of polyclonal antibodies against viral envelope proteins was also studied. Binding studies were performed using dot blot and virus overlay protein binding assays. PPBED was able to bind to membrane proteins from all brain regions, particularly to a protein band of approximately 116 kDa. Neuraminidase treatment of neuronal membrane proteins decreased virus binding; subsequent treatments with β‐galactosidase and manosidase did not increase virus binding inhibition. N‐glycosidase F and trypsin also decreased virus binding, but not the O‐glycanase. Antibodies against viral haemagglutinin‐neuraminidase blocked virus binding more efficiently than antibodies against viral fusion protein. In conclusion: (1) PPBED is able to bind to pig neurones of all brain regions studied and at all ages analysed; (2) a 116 kDa membrane protein containing sialic acid residues with an N‐linked oligosaccharide chain was specifically recognized; (3) PPBED haemagglutinin‐neuraminidase protein seems to play a central role in neural receptor recognition.