Relationship between serum antibody to Bacteroides gingivalis and clinical parameters in periodontitis patients

Prevalence and proportion of black-pigmented Bacteroides species (BPB) in supragingival and subgingival plaque were determined in ten adult periodontitis patients. Serum antibody against Bacteroides gingivalis (Bg) of these patients were tested using ELISA. Clinical parameters (PI, GI, PD, AL) were collected prior to blood withdrawn. Results showed that BPB were detected in all patients. Mean serum anti-Bg IgG level was significantly greater in the patient group than that in healthy control group. Although the sample size was too small to show statistical difference, there was a trend showing the sera anti-Bg IgG level tended to be greater in accordance with the increase of disease severity and BPB%.     

Local and systemic immunoglobulins reactive to Bacteroides gingivalis in rapidly progressive and adult periodontitis

The purpose of this study was to compare the reactivity of the localized immunoglobulins present in the supernatant fluids from in vitro gingival organ cultures of rapidly progressive (RP) and adult periodontitis (AP) patients with those immunoglobulins occurring systemically in the sera of these patients. Tissue excised from periodontitis patients was diagnosed and classified as being either RP or AP and placed in the wells of a multiwell microtiter plate to which RPMI 1640 tissue culture medium was added. Supernatant fluids were harvested and fresh culture medium added to the wells at 24 h intervals. Supernatant fluids and autologous sera from RP and AP patients were tested for reactivity to Bacteroides gingivalis , an organism often implicated in periodontitis, as well as B. asaccharolyticus , a non-oral, non-crossreacting member of the genus Bacteroides , utilizing an ELISA technique. When immunoglobulins present in gingival tissue supernatant fluids were examined, it was found that mean levels reactive to B. gingivalis were significantly greater (p < 0.05) than those to B. asaccharolyticus regardless of patient group. Similarly, the AP group always exhibited significantly (p < 0.05) increased antibody reactivity to B. gingivalis when compared to the RP periodontitis patients. However, when autologous sera from these patients were assayed, no significant differences were observed for immunoglobulin levels reactive to B. gingivalis or B. asaccharolyticus when comparing the AP and RP patient groups, or within the AP group alone. The results indicate that differences exist in the localized immunological responses of RP and AP patients to the periodontopathogen B. gingivalis that are not necessarily reflected in the systemic serum antibody responses of these patients.     

Determination of pseudocholinesterase activity in the gingival crevicular fluid, saliva, and serum from patients with juvenile periodontitis and rapidly progressive periodontitis

By use of a spectrophotometric method, pseudocholinesterase (PCE) activities were determined in gingival crevicular fluid (GCF), saliva, and serum from patients with juvenile periodontitis (JP) and rapidly progressive periodontitis (RPP) and from controls. The PCE activity in the GCF samples was 181 +/- 48 U/L in the JP group, 588 +/- 135 U/L in the RPP group, and 88.5 +/- 29.1 U/L in the control group. Saliva PCE activity levels were 9.1 +/- 1.7 U/L in the JP group, 21.8 +/- 4.5 U/L in the RPP group, and 12.7 +/- 0.8 U/L in the control group. GCF contained a higher PCE activity than saliva but a lower one than that of serum. The RPP group had a significantly higher PCE activity in both the GCF and saliva samples. No significant differences could be found regarding serum enzyme levels. Also, no significant correlations were present between biochemical values and the severity of periodontal disease. GCF may be an important source for the PCE content of saliva. It is suggested that the increased PCE activity seen in RPP patients might be caused by either the direct production of esterases by bacteria or the induction of esterases during periodontal destruction.     

A comparison of antibody levels to Bacteroides gingivalis in serum and crevicular fluid from patients with untreated periodontitis

The levels of IgG antibody to Bacteroides gingivalis were measured in serum and sequential samples of crevicular fluid from healthy and diseased sites in patients with untreated periodontitis using ELISA. All subjects had detectable serum titres but there was a wide variation in titre between subjects. Moderate to strong correlations were found between serum and crevicular fluid levels of IgG. A statistically significant difference was observed between sequential samples of crevicular fluid. There was no difference in the level of specific IgG to B. gingivalis in crevicular fluid between healthy and diseased sites within the same individual.     

Immunodominant antigens of Porphyromonas gingivalis in patients with rapidly progressive periodontitis

We studied 4 isolates of Porphorymonas gingivalis , ATCC 33277, 381, A7A1-28, and W50, to identify major cell surface antigens and select the best strain from which to obtain antigen for a test vaccine. Immunoglobulin G (IgG) titers measured by enzyme-linked immunosorbent assay using whole-cell sonicates as antigen were significantly elevated for the sera of 64 rapidly progressive periodontitis patients relative to sera of 30 normal control subjects for each of the 4 strains studied. Western blots were prepared for all 4 strains and developed using sera from 22 patients and 20 control subjects to identify and determine the frequency of antibody-binding components. The intensity of binding by patient sera was greatest for the 75-kDa and 55-kDa components. The 43-kDa component was also widely recognized. Strains ATCC 33277 and 381 appeared to be antigenically similar. Because of the higher serum antibody titers, the larger proportion of seropositive patients and higher frequency of binding to specific protein components in Western blots, our efforts were focused on strain ATCC 33277. Whole-cell sonicates, proteinase K-digested sonicate, lipopolysaccharide, capsular polysaccharide, and whole-cell protein fractions were prepared and evaluated for anti-genie activity. By dot immunoblot, most of the antibody binding activity was found in the whole-cell protein fraction, with much lesser amounts in lipopolysaccharide and none in capsular polysaccharide. The antibody-binding activity was accessible on the cell surface, since 98.9% of P. gingivalis -specific antibody, including antibody binding to the 43-kDa, 55-kDa and 75-kDa components on Western blot, was removed by whole-cell adsorption. Furthermore, the 43-kDa and 55-kDa but not the 75-kDa component on intact cells were accessible for labeling with 125_I, confirming their cell surface location and accessibility.     

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.     

Prevalence of Bacteroides forsythus, Porphyromonas gingivalis and Actinobacillus actinomycetemcomitans in subgingival microflora of Japanese patients with adult and rapidly progressive periodontitis

BACKGROUND, AIMS: This study investigated the prevalence of Bacteroides forsythus, Porphyromonas gingivalis and Actinobacillus actinomycetemcomitans among various periodontitis patients and healthy individuals in Japan, and correlated it with clinical parameters. METHOD: Subgingival plaque samples were collected from 21 patients with adult periodontitis (AP), 8 with rapidly progressive periodontitis (RPP) and 15 healthy individuals. RESULTS: The frequency detected in culture was as follows: B. forsythus was found in 47.6% of AP sites and in 37.5% of RPP sites. P. gingivalis was identified in 64.3% of AP and 59.4% of RPP sites. A. actinomycetemcomitans was detected in 4.8% of AP and 3.1% of RPP sites. The 3 species were detected in only 2 of the healthy individuals. The proportion of B. forsythus in the total microflora in culture was 0.07% in the healthy group, 4.1% in AP and 2.4% in RPP. The proportions of P. gingivalis were 0% in the healthy group, 18.8% in AP and 16.2% in RPP. The proportion of A. actinomycetemcomitans was very low in all 3 groups. A DNA probe detected B. forsythus in 78.6% of AP and 65.6% of RPP sites, as well as P.gingivalis in 58.3% of AP and 59.4% of RPP sites. A. actinomycetemcomitans was detected in only 1.2% of AP sites. The 3 species were undetectable in the healthy group. CONCLUSIONS: The prevalence and the proportion of B. forsythus and P. gingivalis were significantly correlated with clinical parameters, suggesting that B. forsythus and P. gingivalis are closely related to AP and RPP in the Japanese population.     

Humoral immune responses to Porphymmonas gingivalis and Actinobacillus actinomycetemcomitans in adult periodontitis and rapidly progressive periodontitis

The relationships between various forms of periodontal disease and the avidities of serum antibodies of all 3 immunoglobulin (Ig) classes (IgG, IgM and IgA) to Porphyromonas gingivalis and Actinobacillus actinomycetemcomitans were investigated. Twenty-four patients with untreated adult periodontitis and twelve untreated patients diagnosed as suffering from the early-onset form of periodontitis, rapidly progressive periodontitis, were studied. The latter group were matched for age and sex to healthy controls. Antibody litres were measured and avidity (expressed as molarity) was further assayed using the thiocyanate elution method. Avidity has previously been shown to relate to the biological function of antibody. IgM avidities to P. gingivalis were lower in the rapidly progressive periodontitis group than in the adult periodontitis group (0.54 M vs 0.74 M). IgG avidities tended to be lower in the former than in the latter group (0.58 M vs 0.92 M). In accordance with other workers, seropositivily was defined as an immunoglobulin titre more than twice the median level of control sera. Only 2 of the rapidly progressive periodontitis group were seropositive. Interestingly, the seronegative rapidly progressive periodontitis patients were significantly different (0.53 M vs 0.92 M). The data that patients with various forms of periodontal disease appear to produce antibodies of differing avidity to P. gingivalis suggest that the quality of the humoral immune response to suspected periodontopathogens may have a bearing on the aetiology of periodontal disease.     

Humoral immune responses to Porphyromonas gingivalis before and following therapy in rapidly progressive periodontitis patients.

We have performed studies aimed at elucidating the nature of the humoral immune response in rapidly progressive periodontitis (RPP). We analyzed the sera of 36 periodontally normal subjects and 36 RPP patients for titers and avidities of IgG antibodies reactive with the antigens of Porphyromonas gingivalis using ELISA, prior to and following treatment. We used whole-cell sonicate, purified lipopolysaccharide (LPS), and total extractable protein as plate antigens. Twelve of the patients had antibody titers at least 2-fold greater than the median of the controls and were designated as seropositive. The remaining 24 patients had titers that did not exceed twice the median titer of the controls and were designated as seronegative. For both patient groups, antibody titers were highest when whole-cell antigen was used, intermediate for LPS, and lowest for the protein fraction. Following treatment, median titer for seropositive patients decreased from pretreatment values of 241.7 to 76.5, while median titer for seronegative patients increased from 39.5 to 80.1. Avidities of pretreatment sera from both patient groups for all 3 antigen preparations were lower than the median avidities of the control sera. Avidity significantly increased following treatment to levels greater than those for control sera in both patient groups. Thus, some young adults with severe periodontitis mount a humoral immune response and produce high levels of serum IgG antibodies reactive with antigens of P. gingivalis, while others do not. The antibodies produced are of relatively low avidity, and may therefore be relatively ineffective biologically. Therapy, which greatly reduces antigen load, appears to stimulate production of higher avidity IgG antibodies in both patient groups; in the seropositive group, low avidity antibodies appear to be replaced by antibodies of higher avidity. Both the purified LPS and protein fractions contain reactive antigen(s), although LPS binds more antibody. Our data are consistent with the idea that many RPP patients do not produce protective levels of biologically functional antibody during the course of their natural infection, but they may be stimulated to do so by treatment.     

Serum antibodies against the trypsin-like protease of Bacteroides gingivalis in periodontitis

Antibody levels of IgG, IgA and IgM reactive to the trypsin-like enzyme of B. gingivalis were measured by enzyme-linked immunosorbent assay in serum samples from control subjects with clinically healthy gingiva, patients with adult periodontitis (AP) and patients with rapidly progressive periodontitis (RPP). Both AP and RPP patients had significantly higher levels of specific IgG and IgA antibodies than control subjects with healthy gingiva (p<0.001 in both cases). No significant difference was observed between the specific IgM antibody activity in the AP group compared to the control group. However, a significant difference was seen (p<0.01) for IgM activity in the RPP group compared to controls. The results indicate that the trypsin-like enzyme of B. gingivalis is produced in vivo in sufficient amounts to be immunogenic to the host and particularly in patients with AP or RPP.     

Effects of egg yolk antibody against Porphyromonas gingivalis gingipains in periodontitis patients

Porphyromonas gingivalis gingipains is suspected to be one of the most important causative agents of periodontitis. We postulated that the inhibition of gingipains may reduce the pathogenic nature of P. gingivalis. Anti-P. gingivalis egg yolk antibody (IgY-GP) was isolated from the yolks of hens immunized with purified gingipains. We applied IgY-GP gel subgingivally in periodontitis patients who harbored P. gingivalis in their subgingival flora. Five pairs of contralateral anterior single-rooted teeth were selected. One tooth in each contralateral pair was randomly treated with IgY-GP and subgingival scaling and root planing, whereas the other tooth was treated with SRP alone. The number of P. gingivalis bacteria was assessed by real-time PCR. Bacterial levels were expressed as the percentage of total bacteria. The IgY-GP group had a significant reduction in probing depth. BOP significantly decreased in the IgY-GP group compared to the control group at week 4. The levels of P. gingivalis significantly increased in the control group at week 4, whereas the reduction in the levels of P. gingivalis was sustained in the IgY-GP group. Within the limitations of the present study, IgY-GP was shown to be an effective immunotherapeutic agent in the treatment of periodontitis.     

Juvenile and rapidly progressive periodontitis

Juvenile and rapidly progressive periodontitis are grouped under the heading of early-onset periodontitis. In recent years, much attention has been devoted to studying immunologic factors in early-onset periodontitis. This study was designed to investigate peripheral blood lymphocyte subpopulations, natural killer cells and interleukin-2 receptor positive (IL-2R +) cells in patients with juvenile and rapidly progressive periodontitis. 38 patients with juvenile and 30 patients with rapidly progressive periodontitis, plus 30 normal healthy control subjects were included in the study. Peripheral blood T-lymphocytes, helper T-cells, suppressor T-cells, HLA-DR+ cells, and IL-2R + cells were determined using appropriate monoclonal antibodies and the indirect immunofluorescence method. B-lymphocytes were identified using the direct immunofluorescence technique. Both groups of patients had normal number of total CD3+ T-cells, CD4+ helper T-cells, CD8+ suppressor T-cells, HLA-DR+ cells and IL-2R+ cells. Natural killer cells were found to be significantly elevated in both groups. These findings could contribute to the immunopathogenesis of early-onset periodontitis.     

Proportions and identity of IgA1-degrading bacteria in periodontal pockets from patients with juvenile and rapidly progressive periodontitis

Degradation of immunoglobulin A1 (IgAl) has previously been demonstrated in suspected periodontal pathogens. The present study revealed that a considerable proportion of the microbial flora in periodontal pockets of patients with juvenile periodontitis (median 29%) and rapidly progressive periodontitis (median 27%) was capable of degrading IgAl. Four different types of degradation occurred: Complete degradation of IgAl; extensive degradation leaving the Fc part of the molecule intact; traditional IgAl protease activity yielding intact Fc and Fab fragments, and removal of carbohydrate side chains on the IgAl molecule. Apart from species already known to degrade IgAl. IgAl protease was, for the first time, demonstrated in strains of Veillonella spp. The ability to cleave off carbohydrate side chains was a feature of a wide variety of bacteria belonging to the species Streptococcus sanguis, S. mitior, S. milleri, Veillonella spp., Actinomyces naeslundii. A. viscosus. Arachnia propionica , and Bacterionema matruchotii . The ability of subgingivally colonizing bacteria to degrade IgA may be a factor contributing to aggravation and perpetuation of the inflammatory reaction in the periodontal tissues.     

The capability of Actinobacillus actinomycetemcomitans, Bacteroides gingivalis and Bacteroides intermedius to indicate progressive periodontitis; a retrospective study

This study evaluated the statistical association of Actinobacillus actinomycetemcomitans, Bacteroides gingivalis and Bacteroides intermedius with progressive periodontitis. 146 adults with a history of advanced periodontitis contributed 105 "nonprogressing" and 130 "progressing" periodontal sites. Periodontal disease activity was assessed by radiographic changes in crestal alveolar bone level. The subgingival proportion of the 3 test bacteria was determined by selective and nonselective culturing. The relationship between bacterial proportions and disease progression was evaluated using subgrouping and multiple-regression analyses. All 3 test bacteria had to be considered in order to distinguish nonprogressing and progressing periodontitis with a reasonably high sensitivity. A recovery rate below 0.01% for A. actinomycetemcomitans, 0.1% for B. gingivalis and 2.5% for B. intermedius defined a site with nonprogressing disease with 87% sensitivity and 84% specificity. By utilizing transformed values of the bacterial recovery rates and optimal test criteria determined by multiple regression analysis, it was possible to obtain sensitivities between 83% and 95% and specificities between 86% and 69%. These 3 bacterial species might serve as valuable components of a periodontitis activity test based on microbiological variables.     

Clinical studies of one family manifesting rapidly progressive, juvenile and prepubertal periodontitis

We report clinical, radiographic and historical data on a large family with an unusually high prevalence of periodontitis. The proband, a 20-year-old black male, had the classic features of juvenile periodontitis (JP). His father was periodontally normal, while his mother had lost all her teeth at age 27 because of rapidly progressive periodontitis (RP). In addition to the 13 living children the couple had had 2 miscarriages. Of the children, one had RP, five had JP and two had prepubertal periodontitis (PP). Both maternal grandparents of the proband had become edentulous at an early age, presumably because of early-onset periodontitis. Four of 10 siblings of the proband's mother had early-onset periodontitis. In contrast, the paternal grandparents did not have early-onset periodontitis nor was periodontitis unusually prevalent in the siblings of the proband's father. The pedigree for this family is consistent with, but does not prove, an X-linked dominant pattern of genetic transmission. The natural history of early-onset periodontitis and the relationship among PP, JP and RP are not understood. The fact that the mother of the proband had RP and she had offspring with RP, JP and PP indicates a close relationship among these diseases and argues in favor of a common underlying mechanism. JP was not preceded by PP in the proband nor his affected 21-year-old brother, but one sister had PP, and at age 15 manifested JP. In her case, the alveolar bone around the deciduous molars had been destroyed, but it regenerated as the permanent premolars erupted.(ABSTRACT TRUNCATED AT 250 WORDS)     

A comparison of the reactivity of Eubacterium species with localized and serum immunoglobulins from rapidly progressive and adult periodontitis patients

Immunoglobulins in sera and in supernatant fluids of explant cultures of diseased gingival tissues from 20 rapidly progressive and 20 adult periodontitis sites were tested by an ELISA assay for reactivity with typed strains of Eubacterium alactolyticum, E. brachy, E. limosum and E. nodatum. Immunoglobulins present in tissue culture fluids from both rapidly progressive and adult periodontitis samples reactive with E. brachy and E. nodatum were significantly greater (P less than 0.05) than those reactive with E. alactolyticum or E. limosum. The titers to E. brachy in tissue culture fluids from adult periodontitis were significantly greater (P less than 0.05) than those from rapidly progressive periodontitis; there was no difference in titers to the other three species. The only significant difference in serum titers was that sera from patients with rapidly progressive periodontitis had significantly greater reactivity to E. alactolyticum than did sera from adult periodontitis patients. These data indicate that immunoglobulins in the sera of rapidly progressive and adult periodontitis patients do not necessarily reflect the reactivity of localized immunoglobulins present in the diseased gingival tissue explant culture fluids from these patients.     

快速进展性牙周炎患者外周血单核细胞趋化功能的研究

目的 研究快速进展性牙周炎患者的外周血单核细胞趋化功能是否异常。方法 采用微孔波膜法对10例快速进展性牙周炎患者的外周血单核细胞趋化功能进行测定。结果 发现快速进展性牙周炎患者的外周血单核细胞趋化功能与正常对照者无差异。结论 部分快速进展性牙周炎可能与单核细胞趋化功能缺陷无关。     

Attachment loss and serum antibody levels against autologous and reference strains of Actinobacillus actinomycetemcomitans in untreated localized juvenile periodontitis patients

Abstract Immunological data have been suggested to be a potential tool in the diagnosis, classification and monitoring of periodontal diseases. However, the role of circulating antibodies in periodontal patients is poorly understood. Patients suffering from localized juvenile periodontitis (LJP) are often reported to show high titers of serum IgG antibodies against Aetinobaeillus actinomycetemcomitans (A. actinomycetemcotnitans), but several affected patients do not. Most studies use well-known reference strains of the bacterium for testing against the patients' sera. The aim of the present investigation was to study the relationship between serum IgG antibody levels to autologous A. actinomycetemcomitans strains and clinical attachment loss (CAL). In addition, we wanted to assess the patients’serum titers against 4 well-known reference strains of the bacterium as well as their general potential immunoglobulin response. Intravenous blood samples were taken from 23 LJP patients and 10 healthy individuals, and autologous A. actinomycetemcomitans strains were cultured from 18 of the L.JP patients. CAL was measured at 4 different sites around ail present teeth and assessed as a % of teeth with at least 1 site moderately ≥2<5 mm) or severely (≥5 mm) involved. An enzyme-linked immunosorbent assay (ELISA) was performed to evaluate the serum titers of IgG antibodies to A. actinomycetemcomitans antigens. No significant correlation was found between serum IgG antibody titers to autologous strains and CAL. However, there was a trend that low responders had more moderately affected teeth than had high responders and patients with undetectable A. actinomycetemcomitans levels, which is in agreement with a hypothetically protective role of the antibodies. The total counts of immunoglobulin assessed in all participants showed that the predominant class was IgG and the reference group displayed significantly less (p<0.05) IgG and IgG1 counts than the LJP patients. Both the reaction pattern against reference and autologous strains varied widely. We conclude that the specific antibody response against A. actinomycetemcomitans shows a weak correlation to clinical attachment levels in LJP patients.