Monoclonal antibodies demonstrate multiple epitopes on the O antigens of Salmonella typhimurium LPS.
To investigate the complexity of the antigenic determinants presented on the surface of Salmonella typhimurium, a panel of murine monoclonal antibodies was generated and characterized. Hybridomas specific for S. typhimurium (strain TML, O antigens 1, 4, 12) were produced by immunization with acetone-killed and dried bacteria and standard fusion procedures. In this report, 15 such monoclonal antibodies, all of which bind lipopolysaccharide (LPS) extracted from S. typhimurium, are described. The fine specificity of these antibodies was assessed by examining the differential binding of each antibody to a panel of Salmonella strains, which selectively express different O antigenic determinants. This analysis defined several distinct categories of monoclonal antibodies of varying isotypes. Four anti-O:4-specific antibodies were identified. Two were specific for O:1. One antibody appears to react with the core polysaccharide of S. typhimurium LPS. Several of the monoclonal antibodies recognized LPS determinants that are presumably created by a combination of O antigens. For instance, one bound only to Salmonella strains that expressed both O:1 and O:12, whereas another bound only to those strains which expressed both O:4 and O:12. A group of three antibodies bound to any strain that simultaneously expressed O:1, O:4, and O:12. A distinct group of three monoclonal antibodies also bound strains that expressed O:1, O:4, and O:12, but only when the O:5 antigenic determinant was not present. The latter are, in that respect, S. typhimurium strain TML LPS-specific. The results of this analysis suggest that the epitopes of the S. typhimurium LPS molecule that are recognized by the host are considerably more complex than has been previously indicated by classical serology.
D000818:Animals; D000911:Antibodies, Monoclonal / Q000032:analysis*; D000918:Antibody Specificity; D000942:Antigens, Bacterial / Q000276:immunology*; D001666:Binding Sites, Antibody; D000939:Epitopes / Q000032:analysis*; D005260:Female; D007126:Immunoglobulin Allotypes / Q000032:analysis; D008070:Lipopolysaccharides / Q000276:immunology* / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D019081:O Antigens; D012486:Salmonella typhimurium / Q000276:immunology* / Q000378:metabolism
D000911:Antibodies, Monoclonal; D000942:Antigens, Bacterial; D000939:Epitopes; D007126:Immunoglobulin Allotypes; D008070:Lipopolysaccharides; D019081:O Antigens