Elevated humoral immune response to heat shock protein 60 (hsp60) family in periodontitis patients

The presence of antibodies to the 60-kD human and Porphyromonas gingivalis GroEL hsp60 in the sera and inflamed gingival tissues of periodontitis patients was examined. In order to obtain the antigens, recombinant plasmids carrying human hsp60 and P. gingivalis GroEL genes were constructed and expressed as histidine-tagged recombinant proteins. Immunoreactivities of these proteins were confirmed by MoAbs specific to mammalian hsp60 and cross-reactive with both mammalian and bacterial hsp60. Western blot analysis clearly demonstrated that the number of periodontitis patients showing a positive response to P. gingivalis GroEL was higher than the number of periodontally healthy subjects. Furthermore, anti-P. gingivalis GroEL antibody was detected in all samples of gingival tissue extracts. For human hsp60, a higher frequency of seropositivity was found in the periodontitis patients than in the healthy subjects. In addition, the periodontitis patients demonstrated stronger reactivity compared with the healthy subjects. Quantitative analysis of serum antibodies by ELISA also demonstrated that the levels of antibodies in the sera of patients were significantly higher than those of control subjects. In the gingival tissue extracts, seven out of 10 patients demonstrated a positive response to human hsp60 and tso of these demonstrated strong positivity. Affinity-purified serum antibodies to human hsp60 and P. gingivalis GroEL from selected patients reacted with P. gingivalis GroEL and human hsp60, respectively, suggesting cross-reactivity of antibodies. These results suggest that molecular mimicry between GroEL of the periodontopathic bacterium P. gingivalis and autologous human hsp60 may play some role in immune mechanisms in periodontitis.     

Periodontopathogen- and host-derived immune response in acute coronary syndrome

Owing to molecular mimicry, periodontal pathogen carriage may result in a systemic cross-reactive immune response with the host. The analyses were performed to investigate if serum antibody levels to human heat shock protein 60 (HSP60) are associated with the antibody levels and salivary carriage of two periodontal pathogens, Aggregatibacter actinomycetemcomitans and Porphyromonas gingivalis, as well as with the dental status in patients with acute coronary syndrome (ACS). ACS patients (n = 141) were monitored at baseline when entering to hospital, and after 1 week, 3 months and 1 year. Periodontal status was recorded by dental radiographs, and A. actinomycetemcomitans and P. gingivalis were detected by PCR from saliva at baseline. Serum IgG and IgA antibody levels were determined at all time points. All antibody levels remained quite stable during the follow-up. Serum IgG-class antibody levels to A. actinomycetemcomitans and HSP60 had a strong positive correlation with each other at all time points (r～0.4, P < 0.05). Mean serum IgG antibody levels to HSP60 were significantly higher in the A. actinomycetemcomitans IgG- and IgA-seropositive than in the seronegative patients, but did not differ between the pathogen carriers compared to the non-carriers. HSP60 antibody levels did not differ significantly between the edentulous, non-periodontitis and periodontitis patients. Despite the observed cross-reactivity in the systemic IgG-class antibody response to HSP60 and A. actinomycetemcomitans, the pathogen carriage in saliva or the periodontal status did not affect the HSP60 antibody levels in ACS patients.     

Self-heat shock protein 60 induces tumour necrosis factor-alpha in monocyte-derived macrophage: possible role in chronic inflammatory periodontal disease

Heat shock protein 60 (hsp60) has been increasingly recognized as an important molecule in infectious and autoimmune diseases. We have demonstrated previously that serum antibodies to both human hsp60 and Porphyromonas gingivalis GroEL were elevated in periodontitis patients compared with healthy subjects. In order to clarify the relative importance of hsp60 in the inflammatory response in periodontal disease, the stimulatory effect of human and bacterial hsp60 on the production of tumour necrosis factor-alpha (TNF-alpha) was examined in phorbol myristate acetate (PMA)-stimulated THP-1 cells. As bacterial hsp60s, recombinant P. gingivalis and Actinobacillus actinomycetemcomitans GroEL was used. Human hsp60 but not P. gingivalis or A. actinomycetemcomitans GroEL demonstrated stimulatory activity similar to lipopolysaccharide (LPS) derived from the bacteria. The activity of hsp60 was inhibited by anti-CD14 and anti-Toll-like receptor 4 (TLR4) antibodies, suggesting that both CD14 and TLR4 mediate hsp60 signalling. Immunohistochemical analysis demonstrated that hsp60 is abundantly expressed in periodontitis lesions. Therefore, it is postulated that periodontopathic bacteria stimulate the cells in the periodontium to up-regulate the expression of hsp60, which in turn may stimulate macrophage and possibly other cells to produce proinflammatory cytokines. These mechanisms may be involved in the chronicity and tissue destruction of periodontal disease.     

Accumulation of human heat shock protein 60-reactive T cells in the gingival tissues of periodontitis patients

Heat shock protein 60s (hsp60) are remarkably immunogenic, and both T-cell and antibody responses to hsp60 have been reported in various inflammatory conditions. To clarify the role of hsp60 in T-cell responses in periodontitis, we examined the proliferative response of peripheral blood mononuclear cells (PBMC), as well as the cytokine profile and T-cell clonality, for periodontitis patients and controls following stimulation with recombinant human hsp60 and Porphyromonas gingivalis GroEL. To confirm the infiltration of hsp60-reactive T-cell clones into periodontitis lesions, nucleotide sequences within complementarity-determining region 3 of the T-cell receptor (TCR) beta-chain were compared between hsp60-reactive peripheral blood T cells and periodontitis lesion-infiltrating T cells. Periodontitis patients demonstrated significantly higher proliferative responses of PBMC to human hsp60, but not to P. gingivalis GroEL, than control subjects. The response was inhibited by anti-major histocompatibility complex class II antibodies. Analysis of the nucleotide sequences of the TCR demonstrated that human hsp60-reactive T-cell clones and periodontitis lesion-infiltrating T cells have the same receptors, suggesting that hsp60-reactive T cells accumulate in periodontitis lesions. Analysis of the cytokine profile demonstrated that hsp60-reactive PBMC produced significant levels of gamma interferon (IFN-gamma) in periodontitis patients, whereas P. gingivalis GroEL did not induce any skewing toward a type1 or type2 cytokine profile. In control subjects no significant expression of IFN-gamma or interleukin 4 was induced. These results suggest that periodontitis patients have human hsp60-reactive T cells with a type 1 cytokine profile in their peripheral blood T-cell pools.     

Humoral immunity of older adults with periodontal disease to Porphyromonas gingivalis.

The effect of age on the humoral response to Porphyromonas gingivalis was assessed in groups of adults (25 to 54 years and 55 to 74 years) with periodontal disease and compared with that in age-matched healthy controls. To determine whether there was an antibody response against P. gingivalis, we measured serum antibodies against whole cells of P. gingivalis 381, A7A1-28, and W50. In addition, antibody levels against purified P. gingivalis outer membrane proteins (i.e., the 43-kDa fimbrial protein and a 75-kDa protein) were also evaluated. Elderly subjects showed the same response to P. gingivalis as younger subjects. Immunoglobulin G (IgG) antibodies to both purified proteins were also elevated in both diseased groups as compared with the normal groups. Total serum IgG, IgA, and IgM levels were also determined by an enzyme-linked immunosorbent assay for all four groups. Total serum IgG levels were elevated in older adults with periodontitis and total IgA levels were elevated in both groups of older adults compared with the younger groups of similar disease status. Total serum IgM levels were comparable for the four groups. Antinuclear antibody titers were assessed in the two groups of older adults and were also found to be higher for the group with periodontitis. These studies show that older adults as well as younger adults have markedly elevated specific antibodies of the IgG and IgA classes to antigens of P. gingivalis, a putative pathogen in both groups. Furthermore, older adults with periodontitis have significantly elevated levels of total serum IgG which may possibly be related to higher levels of autoantibodies.     

Relationship of periodontal infection to serum antibody levels to periodontopathic bacteria and inflammatory markers in periodontitis patients with coronary heart disease

Several reports have demonstrated a possible association of periodontal infections with coronary heart disease (CHD) by elevated antibody titre to periodontopathic bacteria in CHD patients compared with non-diseased controls. Although each periodontopathic bacterium may vary in virulence for periodontitis and atherosclerosis, antibody response to multiple bacteria in CHD patients has not been understood fully. Therefore, serum levels of antibody to 12 periodontopathic bacteria together with other atherosclerotic risk markers were compared among 51 patients with CHD, 55 patients with moderate to severe chronic periodontitis and 37 healthy individuals. The antibody response was the most prevalent for Porphyromonas gingivalis, a major causative organism, in CHD as well as periodontitis patients. However, antibody positivity was different between CHD and periodontitis if the response was analysed for two different strains of P. gingivalis, namely FDC381 and Su63. While periodontitis patients were positive for both P. gingivalis FDC381 and Su63, a high frequency of antibody positivity for P. gingivalis Su63 but not for FDC381 was observed in CHD patients. The results indicate that the presence of particular periodontopathic bacteria with high virulence may affect atherogenesis. Identifying the virulence factors of P. gingivalis Su63 may gain insight into the new therapeutic modality for infection-induced deterioration of atherosclerosis.     

Relationship between gingival crevicular fluid and serum antibody titers in young adults with generalized and localized periodontitis.

The objective of the present study was to determine the relationship between concentrations of antibodies in serum and those in gingival crevicular fluid (GCF) of patients with juvenile periodontitis and severe periodontitis. Most antigens used to quantitate antibodies were obtained from a panel of bacteria associated with juvenile periodontitis or severe periodontitis. We further investigated variation in antibody titer among different periodontal sites and the extent to which antibody in GCF is locally derived. Titers of antibody, total immunoglobulin G (IgG), and human serum albumin were determined with sensitive radioimmunoassays. The relationship between serum and GCF antibody was complex. Both person-to-person variability and marked variability within the same subject were found among different sites of similar clinical status. The site-to-site variability was found not only for antibody reactive with periodontal organisms, but also for antitetanus toxoid, total IgG, and even human serum albumin. Generally the variability was in the degree of depression of the level in GCF relative to that in serum. However, anti-Bacteroides gingivalis and anti-Actinobacillus actinomycetemcomitans in GCF often exceeded the level in serum. When antibody titers in serum and GCF were calculated per milligram of human serum albumin, most of the apparent depressions of antibody in GCF disappeared. The ratio of antibody in serum to that in GCF approached unity for all organisms except B. gingivalis and A. actinomycetemcomitans Y4, which were markedly elevated. Furthermore, the level of IgG per milligram of human serum albumin in GCF was about twice the level in serum. We believe that human serum albumin reflects serum contribution to the GCF, and we therefore attribute the increased level of IgG per milligram of albumin in GCF to local synthesis. It appears that anti-B. gingivalis and anti-A. actinomycetemcomitans represent an important portion of this local antibody synthesis, since most seropositive patients with severe or juvenile periodontitis had at least one site elevated, and the magnitudes of the elevations were large in many sites. Those sites yielding elevated antibody exhibited no obvious differences in clinical parameters of probeable depth or attachment level as compared with sites in which antibody levels in GCF were similar to serum levels. Elevated antibody in GCF may relate to changes in disease activity that are not detectable by usual clinical measures.     

Human immune responses to oral microorganisms: patterns of systemic antibody levels to Bacteroides species.

Human systemic antibody levels to oral members of the Bacteroides genus were assessed with an enzyme-linked immunosorbent assay. Antibody levels to B. gingivalis, two homology groups of B. intermedius, B. melaninogenicus, B. denticola, B. loescheii, B. corporis, B. oralis, B. buccae, and B. gracilis were determined in subjects with localized juvenile periodontitis, advanced destructive periodontitis, or adult periodontitis and in normal persons. Significantly elevated serum immunoglobulin G (IgG) antibody levels to B. gingivalis were seen in adult and advanced destructive periodontitis patients. Serum IgM and IgA antibodies were increased in diseased versus normal subjects, whereas negligible levels of serum IgE antibody were detected to this microorganism. Serum IgG antibody levels to B. intermedius were increased in advanced destructive periodontitis patients; however, the frequency of elevated responses were similar among the groups. Extreme antibody levels to the other Bacteroides spp. were occasionally observed in this population. Additionally, all of the elevated levels were found in diseased patients. Distribution analyses of the antibody levels indicated that most patients exhibited a pattern of elevated antibodies to a limited number of the oral Bacteroides spp. The results suggested that elevated systemic antibody levels to oral Bacteroides spp. are more frequently found in periodontal disease patients. These antibody responses presumably reflect a colonization of the patients. The distribution of the responses may indicate the potential pathogenicity of the microorganisms and is consistent with distinctive host-parasite interactions in this disease.     

四环素对侵袭性牙周炎血清抗牙龈卟啉菌IgG抗体亲和力影响的研究

目的：评价结合四环素的牙周非手术治疗对侵袭性牙周炎（AgP）患者牙周附着水平（AL）和血清抗牙龈卟啉菌（Pg）抗体亲和力水平的影响。 方法:研究对象由25名侵袭性牙周炎患者、20名牙周健康者（HS）组成。AgP患者在接受机械性牙周治疗后给服四环素(0.5 g/d)，3个月疗程结束后对病人进行评估。牙周治疗前及治疗3个月、6个月和12个月后常规临床检查、记录牙周附着水平，通过硫氰酸铵分离试验测定血清抗牙龈卟啉菌脂多糖（LPS）的抗体亲和力。 结果:牙周治疗后，牙周附着水平有显著改善(P<0.01);AgP患者血清抗Pg抗体亲和力明显下降(P<0.01)。 结论:结合四环素的机械性牙周治疗能显著降低AgP患者血清抗Pg抗体的亲和力对侵袭性牙周炎可获得满意的疗效。     

Characterization of heat shock protein-specific T cells in atherosclerosis

A role for infection and inflammation in atherogenesis is widely accepted. Arterial endothelium has been shown to express heat shock protein 60 (HSP60) and, since human (hHSP60) and bacterial (GroEL) HSP60s are highly conserved, the immune response to bacteria may result in cross-reactivity, leading to endothelial damage and thus contribute to the pathogenesis of atherosclerosis. In this study, GroEL-specific T-cell lines from peripheral blood and GroEL-, hHSP60-, and Porphyromonas gingivalis-specific T-cell lines from atherosclerotic plaques were established and characterized in terms of their cross-reactive proliferative responses, cytokine and chemokine profiles, and T-cell receptor (TCR) Vbeta expression by flow cytometry. The cross-reactivity of several lines was demonstrated. The cytokine profiles of the artery T-cell lines specific for GroEL, hHSP60, and P. gingivalis demonstrated Th2 phenotype predominance in the CD4 subset and Tc0 phenotype predominance in the CD8 subset. A higher proportion of CD4 cells were positive for interferon-inducible protein 10 and RANTES, with low percentages of cells positive for monocyte chemoattractant protein 1 and macrophage inflammatory protein 1alpha, whereas a high percentage of CD8 cells expressed all four chemokines. Finally, there was overexpression of the TCR Vbeta5.2 family in all lines. These cytokine, chemokine, and Vbeta profiles are similar to those demonstrated previously for P. gingivalis-specific lines established from periodontal disease patients. These results support the hypothesis that in some patients cross-reactivity of the immune response to bacterial HSPs, including those of periodontal pathogens, with arterial endothelial cells expressing hHSP60 may explain the apparent association between atherosclerosis and periodontal infection.     

Multiserotype enzyme-linked immunosorbent assay as a diagnostic aid for periodontitis in large-scale studies

Periodontitis is a common chronic oral infection caused by gram-negative bacteria, including Actinobacillus actinomycetemcomitans and Porphyromonas gingivalis. Periodontitis evokes inflammatory host response locally in the periodontium but also systemically. The systemic humoral antibody response against oral pathogens can conveniently be measured by an immunoassay. The aim of the study was to measure serum immunoglobulin G class antibodies against A. actinomycetemcomitans and P. gingivalis by an enzyme-linked immunosorbent assay (ELISA) in which mixtures of several serotypes of the pathogens were used as antigens to avoid biasing of the results in favor of a particular strain. For A. actinomycetemcomitans the antigen consisted of six strains representing serotypes a, b, c, d, and e and one nonserotypeable strain. In the P. gingivalis ELISA, antigens representing serotypes a, b, and c were used. Serum samples from 90 subjects, including 35 samples from patients with diagnosed periodontitis, 10 samples from periodontally healthy controls, and 45 samples from randomly selected apparently healthy volunteers (referred to as "healthy subjects"), were tested. For both pathogens the antibody levels (means +/- standard deviations) of the patients--xpressed as area under the dilution curve--were significantly higher than those for healthy controls or healthy subjects, with values for A. actinomycetemcomitans and P. gingivalis, respectively, as follows: patients, 22.60 +/- 9.94 mm(2) and 26.72 +/- 11.13 mm(2); healthy controls, 9.99 +/- 3.92 mm(2) and 6.90 +/- 3.38 mm(2); and healthy subjects, 16.85 +/- 6.67 mm(2) and 8.51 +/- 4.23 mm(2). The serotype mixture ELISA is suitable for measuring antibodies against periodontal pathogens in large epidemiological studies in order to evaluate the role of periodontitis as a risk factor for other diseases.     

Serum antibodies to oral Bacteroides asaccharolyticus (Bacteroides gingivalis): relationship to age and periondontal disease.

An enzyme-linked immunosorbent assay microplate method was used for measuring levels of antibody specific for the oral serotype of Bacteroides asaccharolyticus (Bacteroides gingivalis) in serum samples obtained from umbilical cords, infants, children, periodontally normal adults, and edentulous adults. Serum from patients with various periodontal diseases, including adult periodontitis, localized juvenile periodontitis, generalized juvenile periodontitis, post-localized juvenile periodontitis, and acute necrotizing ulcerative gingivitis, were also studied. A positive correlation between increase in age and increase in both prevalence and level of specific antibody in the G, A, and M classes of immunoglobulins was observed. This indicates that antibodies reactive with oral B. asaccharolyticus found in up to 84% of normal adults are natural antibodies, presumably with a protective role. Among the patient groups, those with adult periodontitis were found to have levels of immunoglobulin G antibodies to oral B. asaccharolyticus that were five times higher than the antibody levels found in control subjects. The levels of IgG antibodies to this organism in the other patient groups were comparable to the levels found in the control group. However, 50% of the individuals in the generalized juvenile periodontitis group had high levels of immunoglobulin G antibodies to B. asaccharolyticus, suggesting heterogeneity with respect to immune response in these patients. These results indicate that antibodies to oral B. asaccharolyticus (B. gingivalis) occur at low levels in most normal children and adults and that the rise in titer of the specific antibodies of each major class of immunoglobulins parallels the ontogenic change in serum levels of that isotype. In contrast, there is a marked increase in titer of immunoglobulin G antibodies to oral B. asaccharolyticus in the group of patients with adult periodontitis and in patients with the generalized form of juvenile periodontitis.     

Immunoglobulin G response to subgingival gram-negative bacteria in human subjects

Serum and gingival crevicular fluid from normal healthy adults and patients with periodontitis were screened for immunoglobulin G antibodies to antigens from Bacteroides gingivalis 381, Bacteroides intermedius 24, Bacteroides loescheii ATCC 15930, Fusobacterium nucleatum ATCC 25586, Eikenella corrodens 1073, Actinobacillus actinomycetemcomitans ATCC 29522, and Capnocytophaga sp. strain M-12. Immunoglobulin G antibody titers to the antigens were measured by an enzyme-linked immunosorbent assay. The antibody levels to B. gingivalis in serum and gingival crevicular fluid were significantly higher in the samples from patients with periodontitis than in samples from healthy individuals. Although there were individual differences within patient groups, a positive correlation (P less than 0.01) was found between the serum immunoglobulin G levels to B. gingivalis and the development of periodontitis. The antibodies to F. nucleatum (P less than 0.05), E. corrodens (P less than 0.05), and A. actinomycetemcomitans were slightly higher in patients with periodontitis than in normal subjects. There were no remarkable differences between the two groups in titers to B. intermedius, B. loescheii, and Capnocytophaga sp.     

Lymphotoxin alpha LTA+496C allele is a risk factor for periodontitis in patients with coronary artery disease

Periodontitis and coronary artery disease (CAD) are inflammatory diseases and associated with each other. The major histocompatibility complex (MHC) region carries genes involved in immune response and inflammation. We investigated whether the MHC genes correlate with the presence of periodontitis or with the occurrence of periodontal pathogens in patients with CAD. Blood and saliva samples from CAD patients (n = 106) were collected at the time of hospitalization. Nine MHC genetic markers [human leukocyte antigen (HLA)-A, HLA-B, HLA-DRB1, lymphotoxin alpha (LTA) +253(a/g), +496(C/T), +633(c/g), +724(C/A), C4A and C4B)] were typed. Based on panoramic tomography, patients were categorized into nonperiodontitis and periodontitis groups. Two major periodontal pathogens, Aggregatibacter (Actinobacillus) actinomycetemcomitans and Porphyromonas gingivalis, were cultivated and polymerase chain reaction-amplified from salivary samples. Serum immunoglobulin (Ig)A and IgG antibody levels to these pathogens were measured. In the univariate analysis, LTA+496C allele (OR = 5.29; 95% CI = 2.07-13.51, P = 0.00027), and the occurrence of P. gingivalis in saliva (OR = 4.74; 95% CI = 1.64-13.70; P = 0.002) were more frequent in periodontitis when compared with nonperiodontitis. Similarly, serum IgA antibody level against the pathogen was increased in periodontitis (P = 0.048). In the multiple logistic regression analysis, when a wide range of covariates was included, the LTA+496C allele (OR = 10.87; 95% CI = 3.23-36.60; P = 0.00012) and the elevated serum IgA antibody level against P. gingivalis (OR = 1.56; 95% CI = 1.05-2.30; P = 0.026) remained as significant risk factors for periodontitis. In conclusion, the major finding of this study is that the LTA+496C allele is associated with periodontitis in patients with CAD.     

早发性牙周炎患者血清抗牙龈卟啉菌表面抗原抗体滴度和亲和力的研究(英文)

目的：检测早发性牙周炎患者血清抗牙龈卟啉菌脂多糖IgG抗体的滴度和亲和力，并探讨抗体滴度水平和亲和力之间的相关性。 方法： 受试者为15名早发性牙周炎患者、16名成人牙周炎患者和14名牙周健康者。检测了血清中抗牙龈卟啉菌脂多糖IgG抗体滴度和亲和力。血清IgG抗体滴度采用酶联免疫吸附试验（ELISA）测定；其亲和力通过二乙醇胺分离，ELISA测定。 结果： 早发性牙周炎患者和成人牙周炎患者的血清中抗牙龈卟啉菌脂多糖IgG抗体水平显著高于牙周健康者（P<0.01），而在早发性牙周炎患者和成人牙周炎患者之间无显著差异（P>0.05）。早发性牙周炎患者抗体亲和力显著高于成人牙周炎患者和牙周健康组（P<0.01），而成人牙周炎患者和牙周健康组之间无显著差异（P>0.05）。 结论： 早发性牙周炎患者和成人牙周炎患者的抗体滴度水平和亲和力之间不存在相关性。IgG抗体亲和力可以作为诊断早发性牙周炎的有效参数。     

Functional properties of nonhuman primate antibody to Porphyromonas gingivalis.

The nonhuman primate (NHP) serves as a useful model for examining the host-parasite interactions in Porphyromonas gingivalis-associated periodontal disease. This study determined the influence of NHP sera on (i) the direct killing of P. gingivalis, (ii) P. gingivalis-induced superoxide anion (O2-) release from human polymorphonuclear leukocytes (PMNs), and (iii) the ability of PMNs to bind and phagocytize P. gingivalis. Three types of NHP sera were utilized: (i) normal or baseline sera; (ii) sera obtained after ligature-induced periodontitis; and (iii) sera obtained following active immunization with formalinized P. gingivalis. All assays were performed with or without the addition of human complement. Significantly more (P < 0.01) direct killing of P. gingivalis occurred with immunized sera and complement than with any of the other treatments. The sera from ligature-induced periodontitis NHPs had significantly less (P < 0.03) killing capacity than the baseline sera, which contained natural antibody produced to P. gingivalis colonization. Sera from immunized NHPs were used to opsonize P. gingivalis and caused significantly greater (P < 0.01) levels of O2- release from PMNs. Finally, the sera from immunized NHPs significantly enhanced (P < 0.009) the uptake of P. gingivalis by PMNs, although binding of the bacteria to PMNs was similar among all three serum types. Active immunization of NHPs with P. gingivalis elicited a functional antibody that enhanced direct killing, positively influenced the activation of PMNs, and enhanced the ability of PMNs to phagocytize P. gingivalis. Moreover, antibody produced as a sequela of progressing periodontitis appeared to lack these functions. A wide variability in functional capacity of the sera from individual NHPs, which may contribute to an individual's susceptibility to P. gingivalis-induced disease, was noted. This variability suggested that results from functional tests of serum antibody may aid in predicting host susceptibility to disease and response to therapy.     

Systemic antibody responses to oral microorganisms in the cynomolgus monkey: development of methodology and longitudinal responses during ligature-induced disease.

Systemic antibody responses to oral microorganisms were studied during ligature-induced periodontal disease in a non-human primate (Nhp) model. Methodology was developed using ELISA techniques to assess total IgG and IgM levels in the serum from the Nhp. In addition, an ELISA was developed utilizing affinity-purified anti-human isotype reagents to detect Nhp serum antibody responses to Porphyromonas gingivalis, Actinobacillus actinomycetemcomitans, Prevotella intermedia and Fusobacterium nucleatum. Results showed that the anti-human reagents detected IgG and IgM from Macaca fascicularis with an efficiency of 25-35% and 50-60%, respectively. Following ligation, groups of Nhp were treated with an immunomodulator ("Thymopentin", TP5) or placebo to examine the effect of the T-cell stimulating agent on periodontitis and host responses. No differences were noted in total serum IgG and IgM levels for individual Nhp or between groups when baseline, ligation and treatment intervals were compared. However, following ligation, 8/12 Nhp exhibited significant increases in IgG and/or IgM antibody to P. gingivalis that were coincident with increases in the percentage of this microorganism in the subgingival plaque from the ligated sites. During the treatment phase, the antibody levels in the placebo group continued to increase, while the levels in the TP5-treated group stabilized. The findings in this study indicate that the emergence of a microorganism in the subgingival plaque (P. gingivalis) during the conversion from gingivitis to progressing periodontitis in the Nhp, elicits a systemic antibody response that is specific for the microorganism.     

结合四环素和抗炎剂的牙周治疗对早发性牙周炎抗牙龈卟啉菌抗体水平的影响

目的：评价结合四环素和抗炎剂的牙周治疗对早发性牙周炎抗牙龈卟啉菌抗体IgG水平的影响。 方法： 研究对象由12名早发性牙周炎患者、16名成年牙周炎患者和12名牙周健康者组成。牙周炎患者接受常规牙周治疗，并口服四环素和消炎痛，牙周治疗前及治疗3个月后常规临床检查、记录牙周探诊深度和牙周附着水平及牙周探诊出血指数，并测定治疗前后患者血清抗牙龈卟啉菌抗体IgG滴度的变化。 结果： 牙周治疗后，牙周探诊深度、附着水平和牙周出血指数均有显著改善，血清抗牙龈卟啉菌抗体滴度明显下降。 结论： 结合四环素和消炎痛的机械性牙周治疗对早发性牙周炎可获得满意的疗效。血清抗牙龈卟啉菌抗体水平对评估早发性牙周炎的预后具有重要意义。     

Association of three bacterial species and periodontal status in Chinese adults: an epidemiological approach

Aggregatibacter actinomycetemcomitans, Porphyromonas gingivalis, and Tannerella forsythia are oral pathogens associated with periodontitis. The association between these three bacteria and periodontal disease has been reported in populations of many countries. However, corresponding data in Chinese populations are still lacking. The aim of this study was to detect these pathogens in subgingival plaque collected from 468 subjects with chronic periodontitis in a group of Chinese adults by using a PCR method and to determine the degree of association between the target bacteria and periodontal status based on logistic regression analysis. A. actinomycetemcomitans, P. gingivalis, and T. forsythia were found in 20.5%, 70.7%, and 77.1% of the subjects, respectively. About one-third (36.1%) of subjects had chronic periodontitis. Upon univariate analysis, age, male gender, current smoking status, diabetes, and the presence of A. actinomycetemcomitans or P. gingivalis were positively associated with chronic periodontitis, whereas education and income exhibited inverse associations with chronic periodontitis. Upon multivariate analysis, education, current smoking status, diabetes, and the presence of A. actinomycetemcomitans and P. gingivalis remained significant. The adjusted odds ratios for having chronic periodontitis were 2.5 and 3.4 in subjects positive for A. actinomycetemcomitans and P. gingivalis, respectively. However, no significant association was observed between the presence of T. forsythia and periodontal status. This study assesses the prevalence of periodontal pathogens and chronic periodontitis and the associations with sociodemographic characteristics among this group of Chinese adults. These findings also suggest that PCR should be considered for field oral epidemiologic studies and may be necessary in investigations presenting major logistic challenges.     

The immune responses to human and microbial heat shock proteins in periodontal disease with and without coronary heart disease

The human 60 kDa and microbial 65 kDa heat shock proteins (HSP) have been implicated in the pathogenesis of chronic periodontitis (P) and coronary heart disease (CHD). We have studied four male non-smoking cohorts of 81 subjects, matched for age. Group (a) consisted of a healthy group with minimal gingivitis (n = 18), group (b) were patients with P (n = 23), group (c) patients with CHD and minimal gingivitis (n = 20) and group (d) patients with CHD and P (n = 20). T cells separated from peripheral blood were found to be primed to both microbial HSP65 and human HSP60 but significant CD4, human leucocyte antigen (HLA) class II-restricted proliferative responses were found only with the human HSP60 in patients with P (P < 0.001) and CHD without (P < 0.001) or with (P < 0.00001) periodontitis. Dose-dependent inhibition of T cell proliferative responses was carried out to determine the receptors involved in recognition of HSP60 and HSP65. Monoclonal antibodies to CD14 showed inhibition of T cell proliferation stimulated by both HSP60 and HSP65, consistent with the role of CD14 as a receptor for these HSPs in P and CHD. The toll-like receptor 2 (TLR-) and TLR-4 were then studied and these showed that TLR-4 was recognized by microbial HSP65, whereas TLR-2 was recognised by human HSP60 in both P and CHD. However, a dissociation was found in the HSP60 and TLR4 interaction, as TLR4 appeared to have been recognized by HSP60 in P but not in CHD. The results suggest an autoimmune or cross-reactive CD4(+) class II-restricted T cell response to the human HSP60 in P and CHD. Further studies are required to determine if there is a common epitope within HSP60 that stimulates T cell proliferation in P and CHD.