Antiadhesive properties of a quaternary structure-specific hybridoma antibody against type 1 fimbriae of Escherichia coli.
The relationship between the structure and biological function of type 1 fimbriae of Escherichia coli was investigated using a set of monoclonal antibodies directed against conformation-specific antigenic determinants. Of three monoclonal antibodies tested, only one (clone CD3) prevented adhesion of the vaccine strain to epithelial cells or guinea pig erythrocytes. The antibody produced by CD3, but not that produced by the other two hybridoma clones (AA8 and GG1), precipitated isolated fimbriae by double diffusion in agar gel and was shown to bind in a highly discrete, periodic manner along the length of each of the fimbriae by immunoelectron microscopy. Immunoelectroblots of type 1 fimbrial subunits and polymers electrophoresed in SDS-gels indicated that the antibodies in AA8 and GG1 reacted only with fimbrial monomers (mol wt 17,000), whereas the antibody in CD3 reacted only with polymers of mol wt 102,000 (hexamers) or higher. ELISA inhibition assays demonstrated that dissociated fimbrial subunits lost their reactivity with antibody CD3 but gained reactivity with antibodies AA8 and GG1. Conversely, when allowed to reassemble in vitro in the presence of 5 mM MgCl2, the reassembled fimbriae lost their reactivity with antibodies AA8 and GG1 but regained reactivity with antibody CD3. These results demonstrated that certain antigenic epitopes are dependent on quaternary structural determinants, whereas others are independent of quaternary fimbrial structure and also are inaccessible for antibody binding in fimbriae once they have been assembled. These monoclonal antibodies should prove useful in studies of the structural determinants of the biological function of type 1 fimbriae as well as in studies of fimbrial synthesis, transport, and assembly.
D000268:Adhesiveness; D000818:Animals; D000907:Antibodies, Bacterial / Q000032:analysis*; D000911:Antibodies, Monoclonal / Q000032:analysis / Q000502:physiology; D000937:Antigen-Antibody Reactions; D000942:Antigens, Bacterial / Q000008:administration & dosage / Q000276:immunology; D001667:Binding, Competitive; D000939:Epitopes; D004926:Escherichia coli / Q000276:immunology* / Q000648:ultrastructure; D005260:Female; D010861:Fimbriae, Bacterial / Q000145:classification / Q000276:immunology* / Q000502:physiology; D006386:Hemagglutination Tests; D006825:Hybridomas / Q000276:immunology*; D007074:Immunoglobulin G / Q000032:analysis / Q000145:classification; D051379:Mice; D008807:Mice, Inbred BALB C; D011487:Protein Conformation
D000907:Antibodies, Bacterial; D000911:Antibodies, Monoclonal; D000942:Antigens, Bacterial; D000939:Epitopes; D007074:Immunoglobulin G