Serological characterization of Actinobacillus pleuropneumoniae serotype 2 strains by using polyclonal and monoclonal antibodies.
The antigenic differences between strains of serotype 2 of both biotypes I and II of Actinobacillus pleuropneumoniae were studied by using several serological techniques. Monoclonal antibodies (MAbs) against A. pleuropneumoniae biotype I serotype 2 were produced by fusion of spleen cells of BALb/c mice immunized with whole-cell (WC) suspension with SP2/O-Ag14 murine myeloma cells. Desirable MAbs were selected by enzyme-linked immunosorbent assay (ELISA) using WC as antigen. MAbs MK-7 and MK-10 identified multiple bands of lipopolysaccharide in Western-blot. Treatment of WC with proteinase K and sodium periodate indicated that both MAb binding sites were carbohydrates in nature. In both ELISA and Western-blot, MAbs MK-7 and MK-10 recognized only biotype I serotype 2 isolates. Neither MAb MK-7 nor MK-10 reacted with reference strains of remaining serotypes of A. pleuropneumoniae and other Gram-negative bacteria tested. The results obtained with various serological tests showed that strains of serotype 2 biotype I shared antigenic determinants with strain N-282 of serotype 2 biotype II, but not with strain N-273 of serotype 1 biotype II. It is suggested that data obtained from this study may be helpful in the development of specific serotyping and serodiagnostic reagents of A. pleuropneumoniae strains.
Bouh
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KC
Départment de Pathologie et Microbiologie, Faculté de Médecine Vétérinaire, Université de Montréal, Saint-hyacinthe, Que, Canada.
Mittal
K R
KR
D016977:Actinobacillus pleuropneumoniae / Q000145:classification* / Q000276:immunology / Q000302:isolation & purification; D000372:Agglutination Tests; D000818:Animals; D000906:Antibodies; D000911:Antibodies, Monoclonal; D000942:Antigens, Bacterial / Q000276:immunology; D003378:Counterimmunoelectrophoresis; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D006386:Hemagglutination Tests; D005779:Immunodiffusion; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D011817:Rabbits; D012703:Serotyping / Q000379:methods
D000906:Antibodies; D000911:Antibodies, Monoclonal; D000942:Antigens, Bacterial