Isolation and Characterization of Monoclonal Antibodies to Simian Virus 5 and Their Use in Revealing Antigenic Differences between Human, Canine and Simian Isolates

Abstract

SUMMARYHybridomas secreting monoclonal antibodies to simian virus 5 (SV5) were obtained following immunization of mice with purified preparations of a human isolate (LN) of SV5. Immune precipitation studies showed that these monoclonal antibodies had specificities for the haemagglutinin-neuraminidase (HN), fusion (F), nucleo-, matrix and phospho-(P) proteins of SV5. By use of a radioimmune competition assay the monoclonal antibodies to the HN protein were assigned to four groups, members of which recognized different antigenic sites on the protein. All the anti-HN antibodies and the anti-F antibody neutralized virus infectivity. The 54 monoclonal antibodies obtained were used to determine whether there were antigenic differences between five human, two canine and one simian isolate of SV5. Although most of the monoclonal antibodies reacted with all isolates, a few did reveal antigenic differences in the HN, F and P proteins. Furthermore, analysis by SDS-PAGE showed that while the electrophoretic mobilities of most of the virus polypeptides of these isolates were similar some differences could be detected. In particular the P protein showed the most marked mobility differences between the human, canine and simian isolates. Slight differences in the mobility of the F1 glycoprotein could also be visualized.

Canine respiratory viruses

“…Parainfluenza viruses are classified in the family Paramyxoviridae, subfamily Paramyxovirinae. CPIV is antigenically similar to the simian virus 5 (SV5) and to porcine, bovine, ovine and feline parainfluenza viruses [1,127]. Sequence analysis of the fusion protein-encoding gene has revealed that CPIV has 99.3% nucleotide similarity to porcine parainfluenza virus, 98.5% to SV5 and 59.5% nt to human parainfluenza virus 2 [111].…”

Comparison between Parainfluenza Virus Type 2 and Simian Virus 5: Monoclonal Antibodies Reveal Major Antigenic Differences

“…The procedures used for the isolation of MAbs to PF-2 were the same as those described in detail for the production of MAbs to SV5 (Randall et al, 1987). The nomenclature system for the PF-2 MAbs was similar to that used for the SV5 MAbs i.e.…”

“…1.5 mg/1). At the end of the labelling period the medium was harvested for virus purification and the cells were used to make soluble antigen extracts for the immune precipitation reactions, as described in detail elsewhere (Randall et al, 1987). The proteins present in the immune precipitates and in the partially purified virus preparations were analysed by SDS-PAGE and autoradiography (Randall et al, 1987).…”

“…HNa. MAbs that reacted with the H N protein were tested for their ability to compete with the binding of other MAbs (for method see Randall et al, 1987) with reactivities to the same protein. If no competition between antibodies was noted these antibodies were placed in different groups which were given a number that was incorporated into the name of a particular antibody e.g.…”

“…Thus, although the degree of antigenic relatedness between SV5 and PF-2 is not known it has often been assumed that they are very closely related, and SV5 and PF-2 have even on occasion been classified in the same serotype (Pringle, 1987;Goswami & Russell, 1982). We have previously isolated 54 MAbs to SV5 and used these to show that only minor antigenic variations exist between different simian, canine and human isolates of SV5 (Randall et al, 1987). In contrast, we demonstrate in this paper that there are major antigenic differences between SV5 and PF-2 in that most (55 out of 60) MAbs to these viruses are specific for either SV5 or PF-2.…”