Smoking-related cotinine levels and host responses in chronic periodontitis.

BACKGROUND AND OBJECTIVE: Smoking has been reported to increase the risk of periodontal disease by disrupting the balance of immune responses and tissue repair processes; however, this risk varies among smokers. Cotinine levels in saliva are routinely used to measure the level of smoking, and reflect the quantity of nicotine, and other smoking-related xenobiotics that challenge host systems. This study delineated characteristics of inflammatory mediators in saliva and serum antibody responses to both periodontal pathogens and commensal bacteria in smokers as they related to cotinine levels. MATERIALS AND METHODS: This case-control study (n = 279) examined salivary inflammatory mediator responses [interleukin (IL)-1ß, IL-10, prostaglandin E2, myeloperoxidase and plasminogen activator inhibitor-1], and serum IgG antibody responses to three periodontal pathogens (Aggregatibacter actinomyce-temcomitans, Porphyromonas gingivalis, Treponema denticola) and five commensal oral microorganisms (Veillonella parvula, Streptococcus sanguis, Prevotella loescheii, Actinomyces naeslundii, Capnocytophaga ochracea). RESULTS: The patients were stratified into health (n = 30), gingivitis (n = 55) and periodontitis (n = 184); cotinine levels correlated with reported smoking habits in health, less so with gingivitis, and were not correlated in periodontitis. Of the inflammatory mediators/acute phase proteins, only IL-1ß levels were positively associated (p < 0.001) with the pack years and cotinine levels. As might be predicted, patients with periodontitis smoked more (p < 0.001) and had higher levels of cotinine. IL-1ß and antibody to A. actinomycetemcomitans, P. gingivalis and T. denticola were significantly higher in the patients with periodontitis than either patients with gingivitis or who were healthy. CONCLUSIONS: Generally, antibody to the pathogens and commensals was lower with decreased cotinine levels. Smoking exacerbated differences in both inflammatory mediators and three antibody in periodontal disease compared to healthy subjects.

Smoking and periodontal disease: discrimination of antibody responses to pathogenic and commensal oral bacteria.

Smoking is an independent risk factor for the initiation, extent and severity of periodontal disease. This study examined the ability of the host immune system to discriminate commensal oral bacteria from pathogens at mucosal surfaces, i.e. oral cavity. Serum immunoglobulin (Ig)G antibody reactive with three pathogenic and five commensal oral bacteria in 301 current smokers (age range 21-66 years) were examined by enzyme-linked immunosorbent assay. Clinical features of periodontal health were used as measures of periodontitis. Antibody to the pathogens and salivary cotinine levels were related positively to disease severity; however, the antibody levels were best described by the clinical disease unrelated to the amount of smoking. The data showed a greater immune response to pathogens than commensals that was related specifically to disease extent, and most noted in black males. Significant correlations in individual patient responses to the pathogens and commensals were lost with an increasing extent of periodontitis and serum antibody to the pathogens. Antibody to Porphyromonas gingivalis was particularly distinct with respect to the discriminatory nature of the immune responses in recognizing the pathogens. Antibody responses to selected pathogenic and commensal oral microorganisms differed among racial groups and genders. The antibody response to the pathogens was related to disease severity. The level of antibody to the pathogens, and in particular P. gingivalis, was correlated with disease severity in black and male subsets of patients. The amount of smoking did not appear to impact directly serum antibody levels to these oral bacteria.

Systemic inflammatory responses in progressing periodontitis during pregnancy in a baboon model.

This study tested the hypothesis that pregnant female baboons exhibit increased levels of various inflammatory mediators in serum resulting from ligature-induced periodontitis, and that these profiles would relate to periodontal disease severity/extent in the animals. The animals were sampled at baseline (B), mid-pregnancy (MP; two quadrants ligated) and at delivery (D; four quadrants ligated). All baboons developed increased plaque, gingival inflammation and bleeding, pocket depths and attachment loss following placement of the ligatures. By MP, both prostaglandin E(2) (PGE(2)) and bactericidal permeability inducing factor (BPI) were greater than baseline, while increased levels of interleukin (IL)-6 occurred in the experimental animals by the time of delivery. IL-8, MCP-1 and LBP all decreased from baseline through the ligation phase of the study. Stratification of the animals by baseline clinical presentation demonstrated that PGE(2), LBP, IL-8 and MCP-1 levels were altered throughout the ligation interval, irrespective of baseline clinical values. IL-6, IL-8 and LBP were significantly lower in the subset of animals that demonstrated the least clinical response to ligation, indicative of progressing periodontal disease. PGE(2), macrophage chemotactic protein (MCP)-1, regulated upon activation, normal T cell expressed and secreted (RANTES) and LBP were decreased in the most diseased subset of animals at delivery. Systemic antibody responses to Fusobacterium nucleatum, Porphyromonas gingivalis, Actinobacillus actinomycetemcomitans and Campylobacter rectus were associated most frequently with variations in inflammatory mediator levels. These results provide a profile of systemic inflammatory mediators during ligature-induced periodontitis in pregnant baboons. The relationship of the oral clinical parameters to systemic inflammatory responses is consistent with a contribution to adverse pregnancy outcomes in a subset of the animals.

Systemic endotoxin levels in chronic indolent periodontal infections.

BACKGROUND AND OBJECTIVE: Periodontal disease has been linked with an increased risk of various systemic diseases. A plausible biologic explanation for this link includes the opportunity for oral pathogens to translocate to the circulation as a result of breakdown in integrity of the oral epithelium. This study refined a methodology used to detect endotoxin activity in the serum of subjects with indolent periodontal infections. MATERIAL AND METHODS: The QCL Kinetic Chromogenic Assay (Cambrex) is a kinetic measure of endotoxin activity. Sera from 211 pregnant women with periodontitis enrolled in the Obstetrics and Periodontal Therapy Trial were used to develop the assay further and to evaluate the detection of endotoxin activity that might accompany a low-level bacteremia in chronic periodontitis. RESULTS: We optimized the system to increase the sensitivity and reproducibility of the assay. The refined system was able to detect endotoxin activity in serum at > 0.0125 EU/mL. At baseline (13-16 wk of gestation), 35.5% of the women were positive for endotoxin activity (1.62 +/- 2.21; range: 0.38-15 EU/mL). CONCLUSION: This report describes a sensitive measure of endotoxin activity in serum. The procedure allowed us to document levels of this microbial virulence factor in serum of individuals with indolent infections such as periodontal disease.

Periodontitis in pregnancy: clinical and serum antibody observations from a baboon model of ligature-induced disease.

BACKGROUND: Chronic oral infections that elicit host responses leading to periodontal disease are linked with various sequelae of systemic diseases. This report provides seminal information on the clinical and adaptive immunologic characteristics of a baboon model of ligature-induced periodontitis during pregnancy. METHODS: Female Papio anubis were evaluated for periodontal health at baseline. Ligatures were tied around selected teeth to initiate oral inflammation and periodontitis. Then the animals were bred. At midpregnancy ( approximately 90 days), a clinical evaluation was performed, and additional ligatures were tied on teeth in the contralateral quadrants to maintain progressing periodontitis throughout pregnancy. A final clinical evaluation was done for all experimental teeth after delivery, and ligatures were removed. Serum was collected at all sampling intervals for the determination of antibody levels to a group of 20 oral bacteria. Unligated animals served as controls. RESULTS: At baseline, 16% of animals exhibited minimal plaque and gingival inflammation without periodontal disease. The remaining baboons demonstrated varying levels of inflammation/bleeding, and approximately 20% of the population had periodontal pocketing (>3 mm). Ligated animals expressed increased levels of inflammation and increased probing depths and clinical attachment loss (AL) and could be stratified into multiple subsets postligation based upon changes in clinical parameters at midpregnancy and at delivery. Baboons were categorized into disease susceptibility groups (periodontal disease susceptibility 1 through 4) that described the extent/severity of induced disease during pregnancy. Control animals showed minimal periodontal changes during gestation. Significant differences in serum antibody to multiple oral bacteria were found in animals presenting with periodontitis at baseline and during the 6 months of ligature-induced disease. A significant correlation to antibody to P. gingivalis, which was sustained throughout ligation and pregnancy, was observed with disease presentation. CONCLUSIONS: The clinical presentation at baseline, reflecting the natural history of oral disease in these animals, suggests individual variation that is reflected in the characteristics of the adaptive immune responses to oral bacteria. The variability in the response to ligation with resulting periodontal disease provides a model to document prospectively the relationship between oral and systemic health outcomes.

Differential gender effects of a reduced-calorie diet on systemic inflammatory and immune parameters in nonhuman primates.

BACKGROUND AND OBJECTIVE: Dietary manipulation, including caloric restriction, has been shown to impact host response capabilities significantly, particularly in association with aging. This investigation compared systemic inflammatory and immune-response molecules in rhesus monkeys (Macaca mulatta). MATERIAL AND METHODS: Monkeys on continuous long-term calorie-restricted diets and a matched group of animals on a control ad libitum diet, were examined for systemic response profiles including the effects of both gender and aging. RESULTS: The results demonstrated that haptoglobin and alpha1-antiglycoprotein levels were elevated in the serum of male monkeys. Serum IgG responses to Campylobacter rectus, Actinobacillus actinomycetemcomitans and Porphyromonas gingivalis were significantly elevated in female monkeys. While only the antibody to Fusobacterium nucleatum was significantly affected by the calorie-restricted diet in female monkeys, antibody levels to Prevotella intermedia, C. rectus and Treponema denticola demonstrated a similar trend. CONCLUSION: In this investigation, only certain serum antibody levels were influenced by the age of male animals, which was seemingly related to increasing clinical disease in this gender. More generally, analytes were modulated by gender and/or diet in this oral model system of mucosal microbial challenge.

Effects of age and oral disease on systemic inflammatory and immune parameters in nonhuman primates.

This report evaluated systemic inflammatory and immune biomarkers in a cohort of Macaca mulatta (rhesus monkeys) maintained as a large family social unit, including an age range from <1 year to >24 years. We hypothesized that the systemic host responses would be affected by the age, gender, and clinical oral presentation of the population, each contributing to inflammatory and immune responses that would reflect chronic oral infections. The results demonstrated that the prevalence and severity of periodontitis, including missing teeth, increased significantly with age. Generally, minimal differences in clinical parameters were noted between the genders. Systemic inflammatory mediators, including acute-phase reactants, prostaglandin E(2) (PGE(2)), cytokines/chemokines, and selected matrix metalloproteinases (MMP), demonstrated significant differences among the various age groups of animals. Levels of many of these were increased with age, although PGE(2), RANTES, bactericidal permeability-inducing factor (BPI), MMP-1, and MMP-9 levels were significantly increased in the young group ( approximately 1 to 3 years old) relative to those for the older animals. We observed that in the adult and aged animals, levels of the systemic inflammatory mediators related to gingival inflammation and periodontal tissue destruction were significantly elevated. Serum antibody levels in response to a battery of periodontal pathogens were generally lower in the young animals, <50% of those in the adults, and were significantly related to aging in the cohort. The levels of antibodies, particularly those to Porphorymonas gingivalis, Fusobacterium nucleatum, and Tannerella forsythia, were most significantly elevated in animals with periodontal disease, irrespective of the age of the animal. These results provide a broad description of oral health and host responses in a large cohort of nonhuman primates from very young animals to the aged of this species. The findings afford a base of data with which to examine the ontogeny of host responses at mucosal sites, such as the gingival tissues.

Omega-3 fatty acid effect on alveolar bone loss in rats.

Gingival inflammation and alveolar bone resorption are hallmarks of adult periodontitis, elicited in response to oral micro-organisms such as Porphyromonas gingivalis. We hypothesized that omega (omega)-3 fatty acids (FA) dietary supplementation would modulate inflammatory reactions leading to periodontal disease in infected rats. Rats were fed fish oil (omega-3 FA) or corn oil (n-6 FA) diets for 22 weeks and were infected with P. gingivalis. Rats on the omega-3 FA diet exhibited elevated serum levels of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), documenting diet-induced changes. PCR analyses demonstrated that rats were orally colonized by P. gingivalis; increased IgG antibody levels substantiated this infection. P. gingivalis-infected rats treated with omega-3 FA had significantly less alveolar bone resorption. These results demonstrated the effectiveness of an omega-3 FA-supplemented diet in modulating alveolar bone resorption following P. gingivalis infection, and supported that omega-3 FA may be a useful adjunct in the treatment of periodontal disease.

Cytokine responses to treponema pectinovorum and treponema denticola in human gingival fibroblasts.

Human gingival fibroblasts were challenged with Treponema pectinovorum and Treponema denticola to test three specific hypotheses: (i) these treponemes induce different cytokine profiles from the fibroblasts, (ii) differences in cytokine profiles are observed after challenge with live versus killed treponemes, and (iii) differences in cytokine profiles are noted from different gingival fibroblast cell lines when challenged with these treponemes. Three normal gingival fibroblast cell cultures were challenged with T. pectinovorum and T. denticola strains, and the supernatants were analyzed for cytokine production (i.e., interleukin-1alpha [IL-1alpha], IL-1beta, IL-6, IL-8, IL-10, gamma interferon, macrophage chemotactic protein 1 [MCP-1], platelet-derived growth factor, tumor necrosis factor alpha, and granulocyte-macrophage colony-stimulating factor). Unstimulated fibroblast cell lines produced IL-6, IL-8, and MCP-1. T. pectinovorum routinely elicited the greatest production of these cytokines from the fibroblast cell lines, increasing 10- to 50-fold over basal production. While T. denticola also induced IL-6 and IL-8 production, these levels were generally lower than those elicited by challenge with T. pectinovorum. MCP-1 levels were significantly lower after T. denticola challenge, and the kinetics suggested that this microorganism actually inhibited basal production by the fibroblasts. No basal or stimulated production of the other cytokines was observed. Significant differences were noted in the responsiveness of the various cell lines with respect to the two species of treponemes and the individual cytokines produced. Finally, dead T. pectinovorum generally induced a twofold-greater level of IL-6 and IL-8 than the live bacteria. These results supported the idea that different species of oral treponemes can elicit proinflammatory cytokine production by gingival cells and that this stimulation did not require live microorganisms. Importantly, a unique difference was noted in the ability of T. pectinovorum to induce a robust MCP-1 production, while T. denticola appeared to inhibit this activity of the fibroblasts. While the general cytokine profiles of the fibroblast cell cultures were similar, significant differences were noted in the quantity of individual cytokines produced, which could relate to individual patient variation in local inflammatory responses in the periodontium.

Antigenic specificity of gingival crevicular fluid antibody to Actinobacillus actinomycetemcomitans.

Elevated antibody levels to periodontopathogens in GCF have been identified and used as support for local antibody synthesis in periodontitis. This study examined both cross-sectional and longitudinal GCF samples for the antigenic specificity of antibody in the fluid. GCF samples were collected from each tooth of 27 periodontitis patients infected with A. actinomycetemcomitans. Levels of IgG antibody in the GCF were assessed by means of an ELISA and compared with serum for determination of local elevations. A proportion of those GCF samples that exhibited significantly elevated antibody were examined by Western immunoblotting to outer membrane antigens from A. actinomycetemcomitans. Homologous sera were also examined for comparison of antibody specificities. Of the sites with elevated IgG antibody, 87% were colonized by A. actinomycetemcomitans; however, 46% of sites with A. actinomycetemcomitans infection did not have elevated antibody. Cross-sectional studies identified a 78 to 100% agreement between the antibody specificities in GCF and those in serum. Additionally, patterns of antibody reactivity in both GCF and serum in the subjects were often very distinctive. Longitudinal alterations in GCF antibody were examined in 15 patients through a monitoring interval of up to 2 years and showed a general conservation of specificities. However, 7/15 patients exhibited a definite acquisition of different antibody specificities during the monitoring. These results describe a relationship between elevated local antibody and A. actinomycetemcomitans infection. Furthermore, the antibody specificities in serum appear to reflect generally the local response to this pathogen.

Porphyromonas gingivalis induction of mediator and cytokine secretion by human gingival fibroblasts.

We hypothesized that bacterial viability and strain characteristics of Porphyromonas gingivalis could affect the induction of pro-inflammatory mediator secretion by human gingival fibroblast cultures. Both killed and viable P. gingivalis elicited production of prostaglandin E2, interleukin-1 beta (IL-1 beta), IL-6 and IL-8, although killed P. gingivalis induced generally higher levels, particularly IL-6 and IL-8, compared with the viable bacteria. P. gingivalis strains, which exhibited wild-type levels of trypsin-like protease activity, stimulated human gingival fibroblasts to secrete increased levels of prostaglandin E2 and IL-1 beta, although minimal levels of IL-6 and IL-8 were noted in supernatants from the gingival fibroblast cells. P. gingivalis strains BEI and NG4B19, which have either decreased or undetectable levels of trypsin-like protease, respectively, induced significantly greater IL-6 and IL-8 levels in gingival fibroblast cultures compared with the other strains. The ability of antibody to P. gingivalis to alter human gingival fibroblast production of pro-inflammatory mediators was tested using nonhuman primate antisera. Both immune and nonimmune sera altered the P. gingivalis-generated pattern of mediators from the gingival fibroblasts. We conclude that: (i) viable and killed P. gingivalis were capable of inducing various pro-inflammatory cytokines from human gingival fibroblasts; (ii) strain differences in cytokine induction were noted, and the expression of a trypsin-like protease activity was related to decreased extracellular levels of IL-6 and IL-8; and (iii) the presence of serum, particularly with specific antibody to P. gingivalis, significantly altered human gingival fibroblast cytokine production compared with P. gingivalis alone.

Longitudinal human serum antibody responses to outer membrane antigens of Actinobacillus actinomycetemcomitans.

We hypothesize that serum antibody responses to antigens of a periodontopathogen would vary temporally and that the specificity of these host antibodies would relate to infection and disease activity. To test the hypothesis, we obtained between 6 and 13 serum samples from Actinobacillus actinomycetemcomitans (Aa)-positive (serological and microbiological) periodontitis patients at time points between 18 and 42 months into the study, and evaluated specific antibody responses to outer membrane antigens (OMA) of Aa strain Y4. Sera from these patients detected 22 different OMA. Early-onset (EOP; n=7) and adult (AP; n=11) periodontitis patients responded to 35% and 41% of the OMA, respectively. 2 of 9 sera from healthy subjects detected no antigens and 7/9 sera detected 7% of the OMA (p<0.0001 versus EOP and AP). The frequency of antibody responses to the 17 kDa antigen were similar between diseased, infected patients and the uninfected, normal subjects, suggesting that it may be stimulated as a cross-reactive antigen. Antibody to the 28, 38, and 90 kDa antigens were significantly more common in diseased patients (>90%) versus normal subjects (p<0.01, p<0.002, and p<0.002, respectively) and were unique among diseased, infected patients, which may be indicative of infections with Aa. A 65 kDa antigen showed an increased frequency of reaction in the AP versus the EOP (p=0.01) patients, which exemplified a potential distinction in response to Aa between adult and early-onset disease classifications. Seventeen of 22 OMA could be detected in every sample from at least one patient. Longitudinal samples from seropositive EOP and AP reacted 80-100% of the time with the 17, 28, and 100 kDa antigens. Finally, five antigens of 15, 38, 58, 65, and 79 kDa were detected in 33-83% of the seropositive patients; however, antibody to these OMA reacted variably at different sampling points, suggesting some antigenic response diversity over time.

Systemic acute-phase reactants, C-reactive protein and haptoglobin, in adult periodontitis.

Capture ELISAs with biotinylated monospecific antibodies were developed to detect both C-reactive protein (CRP) and haptoglobin (Hp) in serum of adult periodontitis (AP) patients and normal subjects. Each acute-phase reactant was significantly increased in serum from AP patients with CRP at 9.12 +/- 1.61 mg/l versus 2.17 +/- 0.41 mg/l (P < 0.001) and Hp at 3.68 +/- 0.37 g/l versus 1.12 +/- 0.78 g/l (P < 0.001). Assessment of clinical characteristics of the patients' periodontal disease indicated that CRP and Hp levels were significantly increased in patients with the most frequent disease active episodes (P < 0.02 and P < 0.001, respectively). Longitudinal examination of the Hp levels showed a significant decrease following scaling and root planing (3.68 versus 2.38 g/l; P < 0.01). After a 2-year administration of 50 mg/b.i.d. Flurbiprofen (a non-steroidal anti-inflammatory drug), significantly decreased Hp levels were noted (P < 0.005). CRP levels declined by 35-40% after 1-2 years of treatment with the drug (P < 0.05). The findings indicated that localized infections resulting in increased inflammation and tissue loss in the periodontium elicit systemic host changes manifest by increases in two acute-phase reactants. The conclusions are that either these molecules are formed locally and distributed to the serum, or these presumably localized infections impact upon the systemic components of the host protective responses.

Sequential ELISA for cytokine levels in limited volumes of biological fluids.

We have developed an enzyme-linked immunosorbent assay (ELISA) for the sequential analysis of multiple cytokines in limited volumes of biological fluids, including gingival crevicular fluid (GCF) and fibroblast culture supernatants (CS). GCF and CS samples were assayed for multiple cytokines, including IL-1 beta, IL-6, IL-8, GM-CSF and IFN gamma. Immulon 3 microplates were coated with a monoclonal antibody, and a rabbit polyclonal antibody was used to detect the cytokine of interest. Biological samples (200 microL) were added to an anti-IL-1 beta-coated plate and incubated, and 175 microL of each sample were replicate transferred to an anti-IFN gamma-coated plate containing 25 microL/well of diluent. This was repeated in an identical fashion with sequential replicate transfers to an anti-IL-8-coated and finally an anti-IL-6-coated plate. The cytokine standard was a pooled combination of the recombinant human cytokines that were included in the sequence. The plates were developed using an alkaline phosphatase-conjugated goat anti-rabbit IgG and NPP as the substrate. Individual ELISAs ranged in sensitivity from 30 to 2 pg/0.2 mL, with cross-reactivity between these cytokines of < 1%. Additionally, when the same samples were tested in the sequence ELISA vs. the individual ELISA, there was > 85% correlation between the two assays.

Antigenic diversity in the periodontopathogen, Actinobacillus actinomycetemcomitans.

We have identified a significant level of variation in antibody responses to Actinobacillus actinomycetemcomitans outer membrane antigens (OMA). This study was designed to verify A. actinomycetemcomitans antigenic diversity that could contribute to maintaining this chronic infection despite the host immune response. A. actinomycetemcomitans strains (5 from different patients and 3 the same patient) were cultured and OMA prepared for Western immunoblotting studies. Antigen bands in the OMA were identified using 7 sera obtained from 3 adult periodontitis (AP) and 4 localized juvenile periodontitis (LJP) patients that were documented with elevated A. actinomycetemcomitans antibody and infection. Differences/similarities in the antigen patterns among the A. actinomycetemcomitans strains were assessed using a kappa similarity coefficient. Antigen bands in the A. actinomycetemcomitans strains ranged from 11-150 kDa; however, variation in antigen patterns were noted among the strains. Utilizing the human sera as probes for antigenic diversity, the 5 heterologous strains showed average K=0.23 (p < 0.05), while homologous A. actinomycetemcomitans strains had a K=0.48 (p < 0.02). The A. actinomycetemcomitans OMA were used as probes to describe variability in host antibody and as such presumptive evidence of antigenic diversity in A. actinomycetemcomitans that is colonizing each of the patients. The results showed average K=0.26 (p < 0.05) for the patients when tested against each of the heterologous strains, and K=0.14 when tested against the homologous strains (p > 0.1). Finally, antigen bands of M r 80, 65, 58, 31 and 20 kDa were demonstrated as antigens contributing to the antigenic diversity in A. actinomycetemcomitans.

Host response assessment in recurring periodontitis.

Data derived from periodontitis patients have provided support for a relationship between the distribution of selected members of the periodontopathic microbiota and antibody levels to the intact bacteria in both serum and GCF. These data are consistent with the systemic antibody as a reflection of the host response to an infectious process associated with an episode of disease activity. The purpose of this report is to address the concept that the host antibody responses may help to elucidate the specific etiologic agents and be used to model the risk for future periodontal disease progression in recurring periodontitis. These findings from one study in adult periodontitis patients indicated that elevations in certain antibody specificities are most closely associated with patients exhibiting a risk of disease recurrence. Furthermore, analysis of the frequency of antibody elevations suggested that patients capable of maintaining elevated antibody to these pathogens post-treatment, may be indicative of an individual at less risk. A 2nd investigation was implemented to address questions concerning host-parasite interactions in A. actinomycetemcomitans-associated recurring periodontitis. The results showed distinctive characteristics of local and systemic antibody responses and A. actinomycetemcomitans infection in patients with varying extents of recurrent disease. These longitudinal studies developed evidence for the potential of local and/or systemic antibody responses as indicators of periodontal disease recurrence.

Longitudinal dynamics of infection and serum antibody in A. actinomycetemcomitans periodontitis.

OBJECTS: This report describes one of the first prospective studies delineating the relationship between infection, host antibody responses and disease exacerbations and remissions in a distinct subset of periodontitis patients infected with A. actinomycetemcomitans. DESIGN: The design of this longitudinal study included visits for each patient approximately every 2 months for up to 3 years. SUBJECTS AND METHODS: Subjects (n=51) included 16 adult periodontitis (AP) and 11 early-onset periodontitis (EOP) patients with elevated serum IgG antibody to A. actinomycetemcomitans and infection with this microorganism, 12 AP patients with normal levels of anti-Aa antibody, and 12 normal subjects. MEASUREMENT OUTCOMES: Clinical parameters included a gingival index, plaque index, bleeding on probing, pocket depth, and attachment level. Disease activity was defined as loss of attachment during the monitoring intervals. Serum IgG, IgM and IgA antibody to A. actinomycetemcomitans Y4 (serotype b) was quantitated using an ELISA. Subgingival plaque samples were examined for A. actinomycetemcomitans using colony immunoblotting. Human serum IgG antibody specificities to outer membrane antigens (OMA) of A. actinomycetemcomitans Y4 were determined using Western immunoblotting. RESULTS: A. actinomycetemcomitans-infected AP patients had a higher frequency of teeth infected when compared to the EOP patients. However, the EOP patients exhibited a trend for higher levels of A. actinomycetemcomitans in those teeth that were infected. Active disease patients demonstrated a significantly greater frequency of infected sites, as well as significant elevations in the proportions of A. actinomycetemcomitans. Both EOP and AP groups showed significantly elevated IgG, IgM and IgA antibody to A. actinomycetemcomitans when compared to a periodontally normal group. The level of IgG antibody was significantly elevated in A. actinomycetemcomitans-positive patients with active disease, while IgA antibody was decreased in a number of the active group patients. Plaque samples derived from active sites showed a clear and significant increase in A. actinomycetemcomitans that occurred from 2-6 months prior to the identification of disease activity. Approximately 70% of the active disease patients showed an increase in IgG antibody level by 2-4 months prior to disease activity. Studies of the antigen reactivity patterns of serum IgG indicated that antibody to antigens of 65, 58, 48, 29 and 24 kDa were more frequent in patients who showed active disease, while those patients with the greatest frequency of active disease appeared to show a general decrease in the recognition of the A. actinomycetemcomitans OMA. CONCLUSIONS: It appears that A. actinomycetemcomitans infection relates to a particular type of disease with accompanying antibody responses that reflect periods of active disease. The dynamics of A. actinomycetemcomitans infection and the level and specificity of systemic antibody responses to this pathogen support an important contribution of the immune response to managing this infection.

Effects of aging on antibody avidity to Mycoplasma pulmonis.

Antibody avidity of serum and lung lavage responses was examined in rats to determine aging effects on functional differences in antibody to Mycoplasma pulmonis. Three age groups of animals (weanling, adult and senescent) were immunized with either of two doses of formalinized M. pulmonis as the antigen, or a placebo control. Total immunoglobulin levels and specific antibody responses were examined in serum and lung lavage fluids and subsequently avidity measurements of the same samples were made for the specific antibody to M. pulmonis. The concentration of NH4SCN that dissociated 50% of the antibody was used to determine the avidity index of the serum and lung lavage samples. Total serum IgG and IgA were decreased in the weanling animals when compared to the other two age groups of animals. In contrast, serum IgA levels were substantially increased in senescent animals. Significant increases in serum IgA levels were noted following immunization that was not observed for IgG levels. Substantial increases in both serum antibody and lung lavage antibody were observed in response to immunization with either dose of antigen, but only the lung lavage samples showed both IgG and IgA isotypes differences that were attributable to age. Serum IgG avidity indices gradually increased over time following immunization with higher indices being observed in the weanling animals immunized with the higher M. pulmonis dose. Serum IgA avidity indices also increased over time with no significant differences noted among the age groups. Lung lavage IgG avidity demonstrated slightly higher indices in the weanling animals, while lung lavage IgA avidity showed higher avidity indices in the senescent animals at the higher antigen dose. These data suggest that senescent animals are capable of producing an apparently functional antibody response and that differences noted in increased disease susceptibility in older animals may be attributed to mechanisms other than a dysfunctional humoral immune response at mucosal surfaces.

Antigen specificity of serum antibody in A. actinomycetemcomitans-infected periodontitis patients.

We hypothesized that serum antibody with selected antigen specificities would relate to infection and disease in the patients and, thus, describe the characteristics of potential protective antibody. This study used serum samples from 24 periodontitis patients with subgingival infection and elevated serum IgG antibody to A. actinomycetemcomitans to define the antigenic specificities of IgG, IgM, IgA, and IgG1-4 antibody to A. actinomycetemcomitans strain Y4 outer membrane antigens (OMA). Uniform IgG antibody (> 70% of the patients) was noted to antigens with M(r) of 65, 38, 29, and 17 kDa. Both IgA and IgM specificities reflected those shown for IgG in each patient. IgG1 and IgG2 antibody reacted with several OMA bands in each patient, while IgG3 antibodies were directed to numerous OMA bands in many patients and represented the most broad-based response. The IgG4 response patterns were limited to a few OMA bands. We noted a prominent occurrence of IgG reactions with OMA bands that were characteristic for individual patients. The frequency of responses to OMA of higher M(r) (i.e., > 80 kDa) and to the 34-, 31-, and 24-kDa antigens was positively related to the total IgG antibody levels. Antibody reactive with OMA bands at 65-, 38-, 29-, 17-, 15-, and 11-kDa antigens was detected in patients with few to many teeth infected with A. actinomycetemcomitans. Furthermore, patients with a high percentage of teeth with > or = 6 mm pockets had a decreased frequency of responses to the high-M(r) antigens (i.e. > 90 kDa) as well as to the 58-kDa antigen.(ABSTRACT TRUNCATED AT 250 WORDS)

Human antibody responses to outer envelope antigens of Porphyromonas gingivalis serotypes.

Immunological studies examining the homogeneity of the major antigenic components of P. gingivalis have suggested 3 serotypes and have indicated a limited distribution of the serotypes in an individual patient. These studies prompted us to define the immunodominant antigens and distribution of immune responses to P. gingivalis serotypes. Serum IgG antibody levels in periodontitis patients in the present study were most frequently elevated above the normal subjects when tested against P. gingivalis serotype A (i.e., 33277). Nearly 1/3 of the patients showed significantly elevated antibody to multiple serotypes of the P. gingivalis apparently resulting from cross-reacting antigens. We determined distinctive differences among outer envelope protein and antigen patterns obtained from the three serotypes. Moreover, the results identified considerable similarities in the qualitative and quantitative antigen response patterns among patients to a particular serotype. There was a strong positive correlation between IgG antibody levels (ELISA) and the total level of reactivity determined in the immunoblots, as well as a positive correlation to the proportion of antibody to particular antigens. These findings suggest that responses to these antigens comprised a major portion of the response to the intact microorganism. Additionally, the detection of antibody to particular antigen bands was indicative of early responses to each of the P. gingivalis serotypes. The results of our study indicate that a subpopulation of periodontitis patients develop an extensive serum antibody response often to multiple serotypes of P. gingivalis and may define a patient population with a P. gingivalis disease. Finally, our results indicate a more consistent antigenic composition for P. gingivalis which may enhance the potential for strategies to immunologically interfere with disease caused by this microorganism.