Antigenic Characterization of Measles and SSPE Virus Haemagglutinin by Monoclonal Antibodies

Abstract

SUMMARYHybrid cells secreting monoclonal antibodies directed against the haemagglutinin (H) protein of measles virus (Edmonston) were produced by fusion of mouse myeloma cells with spleen cells derived from immunized mice. Measles antibodies secreted by these cells were tested for their ability to react with measles virus in immunoprecipitation experiments and assays of binding, neutralization, haemagglutination inhibition and haemolysin inhibition. On this basis 21 out of 75 hybridomas could be defined and divided into five functional groups with different properties. However, when tested against other measles virus strains, including those isolated from subacute sclerosing panencephalitis (SSPE) patients, normalized radioimmunoassay (RIA) binding titres showed that the extent to which a given antibody bound could vary greatly with the virus strain examined. Moreover, the biological actions within a group were found to be very heterogeneous, even when high antibody binding titres were observed. These results suggest that different measles virus strains, which are not distinguishable by polyvalent sera, do in fact possess antigenic differences. Furthermore, the functional significance of a given virus epitope may vary from strain to strain. Hybridoma antibodies were also used to demonstrate the occurrence of antigenic changes within the H polypeptide of SSPE virus during the course of a non-productive, persistent infection in vitro.

Effect of Measles Virus Antibodies on a Measles SSPE Virus Persistently Infected C6 Rat Glioma Cell Line

“…Monoclonal antibodies directed against virus haemagglutinin were prepared as previously described (ter Meulen et al, 1981). Cells secreting these antibodies were grown in 800 ml plastic tissue culture flasks under RPMI 1640 medium.…”

“…Only monoclonal antibodies directed against the haemagglutinin of measles virus were effective in modulation. A number of different monoclonal antibodies directed against the H protein have been obtained in our laboratory (ter Meulen et al, 1981 ;Carter et al, 1983). Antibodies were selected which were all known to bind to the membranes of cells persistently infected with SSPE virus Lec or to immunoprecipitate the H protein of this virus (ter Meulen et al, 1981 ;Carter et al, 1983).…”

Antigenicity of the Measles Virus Haemagglutinin Studied by Using Synthetic Peptides

“…The antigenicity of MV H has been analysed by several groups using monoclonal antibodies (ter Meulen et al, 1981 ;Carter et al, 1982;G iraudon & Wild, 1985;Sheshberadaran & Norrby, 1986). These studies have revealed three or four antigenic sites consisting of up to nine epitopes, depending on the methods used.…”

Biological Activity of a Monoclonal Antibody to a Measles Virus Haemagglutinin Epitope Detected Late in Infection

“…Thus far, monoclonal antibodies have been used for the detection of antigenic variation among HA molecules (Birrer et al, 1981 ;Trudgett et al, 1981) and the classification of potential binding sites into groups according to the serological activity and the competitive binding characteristics of their respective monoclonal antibodies (ter Meulen et al, 1981;Carter et al, 1982).…”

“…The production of measles virus HAspecific monoclonal antibodies has facilitated the characterization of these antigenic determinants (McFarlin et al, 1980;Birrer et al, 1981 ;Trudgett et al, 1981 ;Togashi et al, 1981 ;Giraudon & Wild, 1981 ;Bohn et al, 1982). Measles virus HA binding sites have recently been categorized into five biological function groups based on the reactivity of HA-specific monoclonal antibodies in haemagglutination inhibition (HI), haemolysin inhibition and neutralization (NT) assays (ter Meulen et al, 1981). Competitive radioimmunoassay was used by Carter et al (1982) to condense these five functional groups into three binding groups which recognize different, but overlapping, areas of the HA molecule, thus proposing the physical relationships among the binding sites.…”