title: Recognition of antigenic epitopes in lipopolysaccharide and protein from Actinobacillus actinomycetemcomitans by serum antibodies in untreated rapidly progressive periodontitis patients.
abstract: Actinobacillus actinomycetemcomitans has been associated with early-onset periodontitis, including the localized juvenile and rapidly progressive forms. The immunodominant antigens of A. actinomycetemcomitans recognized by rapidly progressive periodontitis patients remain unidentified. Sera from 22 patients with rapidly progressive periodontitis and 20 periodontally normal subjects were tested by enzyme-linked immunosorbent assay (ELISA) for immunoglobulin G antibodies to whole-cell sonicate, protein, purified lipopolysaccharide and lipopolysaccharide fractions of A. actinomycetemcomitans. The median titers of rapidly progressive periodontitis patients and control subjects to whole-cell sonicate were 25.0 and 14.5 ELISA units, respectively (not significantly different). Binding of antibody from patient sera occurred to both the lipopolysaccharide and the protein fractions, with greater binding to lipopolysaccharide than to protein. We show for the first time that patient sera contain antibodies that bind specifically to antigenic epitopes in lipid A and in the core carbohydrate of lipopolysaccharide that were previously considered to be inaccessible and unavailable, as well as to epitopes in the O side chains. Sera manifesting antibody titers 2-fold or greater than the median titer for control sera were judged to be seropositive. More patients were seropositive for lipid A than for any of the other antigen preparations studied, and the median titer for patient sera to lipid A but to none of the other purified lipopolysaccharide fractions was significantly elevated relative to control values. Of 22 patients, 10 were seropositive to whole-cell sonicate, 7 to protein, 8 to lipopolysaccharide, 7 to the high-molecular-weight lipopolysaccharide-polysaccharide fraction rich in O side chains, and 16 to lipid A. The core carbohydrate did not adhere to the test plate surface, and this precluded ELISA measurements. However, when the core carbohydrate was used in the ELISA inhibition assay, it reduced antibody binding to lipopolysaccharide-coated plates by up to 45%, thereby demonstrating antibody binding to core carbohydrate. The core carbohydrate fraction from the Re mutant of Salmonella minnesota known to contain no O-side chains also inhibited binding of specific antibody to plates coated with A actinomycetemcomitans lipopolysaccharide. Overall, there was extreme variation in responses among patients to the various antigen preparations, with no single pattern dominating. Lipopolysaccharide and its components appear to be the immunodominant epitopes, since most rapidly progressive periodontitis patients are seropositive for lipopolysaccharide and/or its components and they have titers relative to those for proteins.
authors.affiliation: Department of Periodontics, School of Dentistry, University of Washington, Seattle, USA.
authors.forename: J G
authors.initials: JG
authors.lastname: Ou
authors.forename: B
authors.initials: B
authors.lastname: Bainbridge
authors.forename: K
authors.initials: K
authors.lastname: Gu
authors.forename: T J
authors.initials: TJ
authors.lastname: Sims
authors.forename: C W
authors.initials: CW
authors.lastname: Whitney
authors.forename: R P
authors.initials: RP
authors.lastname: Darveau
authors.forename: H A
authors.initials: HA
authors.lastname: Chen
authors.forename: L S
authors.initials: LS
authors.lastname: Houston
authors.forename: R C
authors.initials: RC
authors.lastname: Page
mesh_terms: D000328:Adult; D016976:Aggregatibacter actinomycetemcomitans / Q000737:chemistry* / Q000276:immunology; D000907:Antibodies, Bacterial / Q000097:blood / Q000276:immunology*; D000918:Antibody Specificity; D000942:Antigens, Bacterial / Q000276:immunology*; D001426:Bacterial Proteins / Q000276:immunology*; D001667:Binding, Competitive; D016022:Case-Control Studies; D003429:Cross Reactions; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D006801:Humans; D016056:Immunodominant Epitopes; D007074:Immunoglobulin G / Q000097:blood / Q000276:immunology; D008050:Lipid A / Q000276:immunology; D008070:Lipopolysaccharides / Q000737:chemistry / Q000276:immunology*; D008297:Male; D008970:Molecular Weight; D010518:Periodontitis / Q000276:immunology* / Q000382:microbiology; D018709:Statistics, Nonparametric
chemical_list: D000907:Antibodies, Bacterial; D000942:Antigens, Bacterial; D001426:Bacterial Proteins; D016056:Immunodominant Epitopes; D007074:Immunoglobulin G; D008050:Lipid A; D008070:Lipopolysaccharides