Physical and antigenic heterogeneity in the flagellins of Listeria monocytogenes and L. ivanovii.
Listeria monocytogenes serotypes 4a, 4b and 7, and L. ivanovii, all grown at 20 degrees C, were negatively stained and examined by electron microscopy. Crude extracts of the cell surface of L. monocytogenes serotypes 1/2b, 3b, 3c, 4a, 4b, 4d and 7 and of L. ivanovii (all grown at 20 degrees C) were examined by SDS-PAGE and Western blotting using (i) affinity-purified polyclonal monospecific antibody, and (ii) monoclonal antibody, each raised against 29 kDa flagellin of serotype 4b. No flagella were seen on serotype 7 by electron microscopy and no flagellin was detected in crude cell surface extracts of serotype 7 either in silver-stained gels or in Western blots. The monospecific polyclonal antibody detected flagellins of approximate molecular mass 29 kDa in each of the seven flagellate strains including L. ivanovii. The monoclonal antibody detected 29 kDa flagellin in serotypes 1/2b, 3b, 4a, 4b and 4d, but not the flagellins of serotype 3c or L. ivanovii, which had a slightly lower molecular mass. Following prolonged electrophoresis of crude flagellar extracts the 29 kDa complex was resolved into three closely migrating bands. In a heterologous system using serotype 1/2b crude flagellar extract, all three bands were detected using the polyclonal antibody whereas only two bands were detected by the monoclonal antibody. It is concluded that polyclonal anti-flagellin antibodies are not useful tools with which to distinguish serotypes of L. monocytogenes sensu lato in immunoblotting, but that differences can be determined using a monoclonal antibody directed against particular components of the flagellar complex. These differences did not fully correspond to those anticipated from results of agglutination tests.
Peel
M
M
Moredun Research Institute, Edinburgh, UK.
Donachie
W
W
Shaw
A
A
D000942:Antigens, Bacterial / Q000032:analysis*; D001426:Bacterial Proteins / Q000032:analysis*; D015153:Blotting, Western; D004591:Electrophoresis, Polyacrylamide Gel; D005407:Flagella / Q000648:ultrastructure; D005408:Flagellin / Q000032:analysis* / Q000276:immunology; D008087:Listeria / Q000032:analysis* / Q000276:immunology / Q000648:ultrastructure; D008089:Listeria monocytogenes / Q000032:analysis* / Q000145:classification / Q000276:immunology; D012703:Serotyping
D000942:Antigens, Bacterial; D001426:Bacterial Proteins; D005408:Flagellin