Effect of periodontal therapy on specific antibody responses to suspected periodontopathogens.

The effects of clinically successful periodontal therapy were studied in juvenile periodontitis (JP) and rapidly progressive periodontitis (RP) patients and compared with periodontally healthy subjects (HS). Serum samples were obtained in 35 HS prior to the study and in 12 of these subjects 3-4 years later. Serum samples were obtained from 50 JP patients initially, 9 subjects immediately following surgical therapy and 29 of these subjects 3-4 years later. RP patients provided 46 initial serum samples, 9 following therapy and 27 samples 3-4 years later. Antibody levels were determined utilizing a standardized enzyme-linked immunosorbent assay with Bacteroides gingivalis, B. ochracea, Fusobacterium nucleatum and Actinobacillus actinomycetemcomitans serving as antigens. The JP patients showed an initial rise in antibody levels immediately following therapy followed by a significant decrease in antibody levels 3 to 4 years later. The RP patients did not show an early change in antibody levels but by 3 to 4 years post-therapy, antibody levels had significantly decreased. However, during this study, the antibody levels of JP and RP patients remained significantly higher when compared with HS patients.

Specificity of monoclonal antibodies reactive with Fusobacterium nucleatum: effect of formalin fixation.

Monoclonal antibody specific for Fusobacterium nucleatum was reacted with untreated and formalin fixed F. nucleatum cells by an enzyme-linked immunosorbent assay (ELISA) and by indirect immunofluorescence. Treatment of bacterial cells with formalin destroyed the antigenic determinant responsible for reactivity with this monoclonal antibody in both assays. Formalin fixation had no effect on hemagglutination activity (HA) of F. nucleatum cells or reactivity with polyvalent rabbit antiserum in double diffusion in agar. Scanning electron microscopy demonstrated that formalin fixation did not affect binding of F. nucleatum cells to microtiter plates. When developing monoclonal antibodies to be used as diagnostic reagents, the antigenic form utilized for immunization should be identical to the antigenic form which will eventually be used in the diagnostic assay.

Reaction of human sera from juvenile periodontitis, rapidly progressive periodontitis, and adult periodontitis patients with selected periodontopathogens.

The levels of serum antibody reactive to selected periodontopathogens were determined in 182 clinically characterized patients: 35 healthy control, 50 juvenile periodontitis, 42 adult periodontitis and 55 rapidly progressive periodontitis. Reactive antibody levels were determined using an enzyme-linked immunosorbent assay with whole cell preparations of Bacteroides gingivalis, Capnocytophaga (Bacteroides) ochraceus, Fusobacterium nucleatum and Actinobacillus actinomycetemcomitans (Y-4) serving as antigens. Increased reactivity to B. gingivalis and F. nucleatum was observed in all three disease groups studied while antibody reactive to A. actinomycetemcomitans was increased in juvenile and rapidly progressive periodontitis. Antibody levels reactive to C. ochraceus in healthy subjects did not differ from those observed in any disease patient groups. Possible implications in the etiology and progression of the diseases coupled with environmental changes which occur in the econiche of the periodontal pocket are described.

Biologic activity of type I and type II Fusobacterium nucleatum isolates from clinically characterized sites.

Fusobacterium nucleatum is a microorganism commonly cultured from periodontal disease sites. F. nucleatum isolates (120) were obtained from subgingival plaque samples taken from 27 clinically characterized sites using a selective culture medium. All isolates were verified by morphology, Gram-stain reactions, oxygen tolerance and biochemical reactions. A total of eight clinical isolates and two typed strains were used for further evaluation. In this study, there was no relationship found between GI and probing depth or between probing depth and frequency of isolation of Type I or Type II F. nucleatum colonies. There was a significant increase in isolation of Type II colonies with a GI of 2 (P less than 0.0001). All isolates tested shared lines of identity by double diffusion in agar and displayed similar ability to hemagglutinate sheep erythrocytes and a reduction in this hemagglutination activity by previous exposure to 50 mM D-galactose. All isolates tested showed similar protein patterns as determined by polyacrylamide gel electrophoresis. By the methods used, no differences were detected between Type I and Type II F. nucleatum; however, there was a statistically significant increase in Type II isolates with increasing levels of gingivitis.