Different responses in B cells induced by Porphyromonas gingivalis and Fusobacterium nucleatum.

A phenotypic study had shown that gingival B cells respond differently to two periodontopathic bacteria, Porphyromonas gingivalis and Fusobacterium nucleatum. Further investigation now shows a reduction in the percentage of Ki-67 + T cells in cultures of gingival and peripheral blood mononuclear cells stimulated with P. gingivalis for 3 and 6 days, respectively, but no suppression of Ki-67 expression in B cells in response to either P. gingivalis or F. nucleatum. Depletion studies of cultures of peripheral blood mononuclear cells showed that in the absence of CD4 cells, the percentage of CD19+ and CD20+ B cells stimulated with P. gingivalis increased after 6 days whereas depletion of CD8 cells resulted in a rise in the percentage of F. nucleatum- and P. gingivalis-stimulated B cells, although this was not significant in the case of P. gingivalis. Specific antibody to P. gingivalis and F. nucleatum was found in culture supernatants of gingival but not of peripheral blood mononuclear cells, indicating a possible higher frequency of antigen-specific B cells in periodontal lesions. IgG was the predominant isotype in both gingival and control peripheral blood cultures, followed closely by IgA in gingival cultures. F. nucleatum stimulated higher levels of Ig in cultures of peripheral blood mononuclear cells than P. gingivalis or cells cultured in medium only, whereas in gingival cell cultures, stimulation by P. gingivalis appeared to result in higher levels of IgG. Also Ig was present at day 3 in gingival cultures, whereas in the blood cell cultures, Ig was only detected at day 6, further suggesting a degree of activation of of gingival B cells.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effect of initial periodontal therapy on the frequency of detecting Bacteroides forsythus, Porphyromonas gingivalis, and Actinobacillus actinomycetemcomitans.

BACKGROUND: Porphyromonas gingivalis, Bacteroides forsythus, and Actinobacillus actinomycetemcomitans have been described as periodontopathic bacteria, and their presence in subgingival pockets can lead to development of periodontal disease. Until now, clinical parameters have been used to evaluate the effect of conventional periodontal treatment without microbiological parameters. The present study examined the microbiological effects of initial periodontal therapy using DNA probes and the polymerase chain reaction (PCR). METHODS: Twenty-six patients with periodontitis, 10 males and 16 females, were given instructions regarding oral hygiene, then thoroughly treated by conventional scaling and root planing. Bacterial samples were collected on paper points from 4 sites per patient at baseline and after initial therapy (total: 104 sites). Clinical parameters including probing depth, attachment level, and bleeding on probing were also recorded for each site at baseline and after therapy. A DNA probe kit was used to monitor the frequency of B. forsythus, P. gingivalis, and A. actinomycetemcomitans, the last of which was identified by PCR. RESULTS: At baseline, B. forsythus was the bacterium most frequently detected. DNA probe analysis also showed that more than half of the sites were colonized by both B. forsythus and P. gingivalis. Initial therapy resulted in significant clinical improvement such as significant reduction in the frequency of B. forsythus and P. gingivalis detected using the DNA probe. A. actinomycetemcomitans was difficult to detect using the DNA probe, but PCR indicated that levels of A. actinomycetemcomitans did not significantly decrease. CONCLUSIONS: These results indicate that initial conventional therapy can eliminate B. forsythus and P. gingivalis, but not A. actinomycetemcomitans. When levels of these bacteria decreased to below-detectable levels, clinical improvement was significant. These results indicate that monitoring levels of these three periodontopathic bacteria may render periodontal therapy more effective and accurate.     

Inhibitory effects of macrocarpals on the biological activity of Porphyromonas gingivalis and other periodontopathic bacteria

BACKGROUND/AIMS: Macrocarpals, which are phloroglucinol derivatives contained in eucalyptus leaves, exhibit antimicrobial activity against a variety of bacteria including oral bacteria. This study examined effects of macrocarpals A, B, and C on periodontopathic bacteria, especially Porphyromonas gingivalis. METHODS: Macrocarpals A, B, and C were purified from a 60% ethanol-extract of Eucalyptus globules leaves. To investigate antibacterial activity, representative periodontopathic bacteria were cultured in media with or without various amounts of macrocarpals; subsequently, the optical density at 660 nm was measured. Macrocarpal inhibition of P. gingivalis Arg- and Lys-specific proteinases was assessed by spectrofluorophotometric assay and sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis. The effect of macrocarpals on P. gingivalis binding to saliva-coated hydroxyapatite beads was examined with (3)H-labeled P. gingivalis. RESULTS: Growth of P. gingivalis was inhibited more strongly than growth of Prevotella intermedia or Prevotella nigrescens and Treponema denticola by macrocarpals, however, Actinobacillus actinomycetemcomitans and Fusobacterium nucleatum were much more resistant. Macrocarpals inhibited P. gingivalis Arg- and Lys-specific proteinases in a dose-dependent manner. The enzyme-inhibitory effect of macrocarpals was confirmed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis in which hemoglobin degradation by P. gingivalis proteinase was inhibited by macrocarpals. P. gingivalis binding to saliva-coated hydroxyapatite beads was also strongly attenuated by macrocarpals. CONCLUSIONS: Macrocarpals A, B and C demonstrated antibacterial activity against periodontopathic bacteria. Among tested bacteria, P. gingivalis displayed the greatest sensitivity to macrocarpals; additionally, its trypsin-like proteinase activity and binding to saliva-coated hydroxyapatite beads were inhibited by macrocarpals. These results indicate that eucalyptus leaf extracts may be useful as a potent preventative of periodontal disease.     

Phenotypic analysis of B-cells extracted from human periodontal disease tissue

B-cells extracted from periodontal disease tissue were analyzed for the presence of activation markers using a range of monoclonal antibodies. In adult periodontitis (AP), 6% of B-cells expressed the IL-2 receptor (CD25) compared with 1-2% in peripheral blood and healthy or marginal gingivitis (H/MG) gingival B-cells. There was also an increase in the mean percentage of IgD-positive B-cells and a decrease in CD21 and CD22 expression. In both AP and H/MG lesions, 20-22% of the B-cells expressed CD23 compared with less than 5% in peripheral blood. As B-cells are activated by day 3 in culture and start differentiating into immunoglobulin-secreting cells by day 6, B-cell phenotypes were assayed at these times in this study. Following stimulation with the periodontopathic bacterium Porphyromonas gingivalis, the expression of CD23, CD21 and CD22 on B-cells extracted from AP lesions remained relatively constant over the 6-d culture period. However, with Fusobacterium nucleatum stimulation, there was a significant decrease in CD23, CD21 and CD22 expression after 3 d in culture, which corresponds to the activation time for B-cells. These results show that B-cells extracted from periodontal disease tissue display a range of activation markers and on stimulation, demonstrate differing responses to individual periodontopathic bacteria.     

The role of cytokines in a Porphyromonas gingivalis-induced murine abscess model

INTRODUCTION: Porphyromonas gingivalis is an important periodontopathic bacterium that is strongly associated with periodontal disease and is part of human dental plaque. Periodontal disease results from the interaction of the host with bacterial products, and T-cell-derived cytokines remain critical in the immunoregulation of periodontal disease. METHODS: The aim of this study was to examine the role of T helper type 1 [interleukin-12p40 (IL-12p40), interferon-gamma, tumour necrosis factor (TNF)] and type 2 (IL-4, IL-10) cytokines in the immune response to a subcutaneous challenge with P. gingivalis using a well-established murine abscess model, in genetically modified cytokine-specific knockout mice. RESULTS: IL-12p40(-/-) mice exhibited more advanced tissue destruction and a reduced inflammatory cell infiltrate after subcutaneous P. gingivalis challenge. Deficiency of IL-4 or IL-10 did not result in increased susceptibility to P. gingivalis-mediated tissue destruction. Furthermore, TNF deficiency appeared to reduce local tissue destruction. Interestingly, serum-specific antibodies suggested a strong T helper type 2 response. CONCLUSION: The results of our study indicate an important role for IL-12 in a primary P. gingivalis subcutaneous challenge.     

Effects of periodontopathic bacteria on IL-1 and IL-1 inhibitor production by human polymorphonuclear neutrophils

The effect of heat-killed periodontopathic bacteria on the production of interleukin-1 (IL-1) and an IL-1 inhibitor by human polymorphonuclear neutrophils (PMN's) was examined. Peripheral blood was obtained from 18 healthy volunteers and the PMN's were separated using dextran sedimentation and Ficoll-Paque density gradient centrifugation. The PMN's (5 x 10(5) cells/well) were cultured in serum-free media with or without heat-killed periodontopathic bacteria. Four gram-negative periodontopathic bacteria were used; Bacteroides gingivalis FDC 381, Bacteroides forsythus FDC 338, Actinobacillus actinomycetemcomitans Y4 and Fusobacterium nucleatum FDC 263. The non-oral Fusobacterium mortiferum ATCC 25557 was used as a control organism. IL-1 activity was assayed using thymocyte proliferation. The non-oral organism F. mortiferum stimulated IL-1 production by PMN's, in contrast none of the periodontopathic bacteria stimulated IL-1 release although the bacteria themselves had an IL-1 enhancing effect. Following fractionation of the periodontopathic bacteria stimulated PMN supernatants, an IL-1 inhibitory fraction was identified. These results may illustrate a further mechanism by which periodontopathic bacteria may evade the protective effect of PMN's and may also suggest a regulatory role for PMN's in chronic inflammatory periodontal disease.     

Induction of apoptosis in human gingival fibroblasts by a Porphyromonas gingivalis protease preparation.

Proteases produced by Porphyromonas gingivalis are believed to contribute to the pathogenesis of periodontal diseases. Here the cytotoxic effects of a purified preparation of a P. gingivalis protease with trypsin-like specificity were tested on human gingival fibroblasts in vitro. The active protease induced apoptotic cell death in the fibroblasts, as indicated by DNA fragmentation and the expression of 7A6 antigen. Thus, the production of proteases by periodontopathic bacteria could be an important factor in the induction of apoptosis of host cells in the aetiology of periodontal diseases.     

Transmission of periodontal disease-associated bacteria from teeth to osseointegrated implant regions

PURPOSE: The presence of periodontopathic bacteria is a risk factor for peri-implantitis. The present study examined colonization by periodontopathic bacteria and their transmission from periodontal pockets to osseointegrated implant sulcus. MATERIALS AND METHODS: Plaque samples were collected from 105 sites in the 15 patients who participated in the study. Colonization by these bacteria was examined by polymerase chain reaction (PCR) and culture. The transmission of periodontopathic bacteria from periodontal sites of natural teeth to the implant sulcus was analyzed by pulsed field gel electrophoresis (PFGE). RESULTS: The PCR detection rates of Porphyromonas gingivalis, Prevotella intermedia, Actinobacillus actinomycetemcomitans, Bacteroides forsythus, and Treponema denticola were 80.0%, 53.3%, 46.7%, 60.0% and 40.0%, respectively. Colonizations by P gingivalis and A actinomycetemcomitans were statistically correlated with periodontal pockets and implant sulcus regions (P < .01). The PFGE patterns of the P gingivalis strains isolated from each patient were identical, but differed from those from other patients. The PFGE patterns of P intermedia strains were identical in 2 out of 3 patients. DISCUSSION: These analyses indicated that there appeared to be transmission of P gingivalis and P intermedia from the periodontal pocket to the peri-implant region. CONCLUSION: Elimination of these periodontal pathogens from the patient's oral cavity before administering dental implant treatment may inhibit colonization by these pathogens and reduce the risk of peri-implantitis.     

Effects of Porphyromonas gingivalis antigens and proinflammatory cytokines on human coronary artery endothelial cells

OBJECTIVE: Individuals with periodontitis have been cited as having a significantly increased risk of developing coronary heart disease. Although accumulating evidence suggests that periodontal infection is involved in the development and progression of atherosclerosis, the underlying mechanisms remain to be elucidated. In the present study, we examined how periodontal infection could contribute to endothelial dysfunction. METHODS: Human coronary arterial endothelial cells were stimulated with tumor necrosis factor (TNF)-alpha and interleukin (IL)-1beta, both of which are reported to be elevated in the serum of periodontitis patients. Cells were also stimulated with lipopolysaccharide, outer membrane protein and heat shock protein 60 derived from Porphyromonas gingivalis, a representative periodontopathic bacterium which is known to stimulate myeloid cells. RESULTS: Although TNF-alpha and IL-1beta, at concentrations a little higher than those in sera of periodontitis patients, up-regulated the expression of intercellular adhesion molecule-1 and vascular cell adhesion molecule-1, P. gingivalis antigens had only a slight stimulatory effect. CONCLUSION: Experiments in which the total pathogen burden is considered, rather than a single species of bacteria, would increase our understanding of the contribution of which periodontal infection to atherogenesis.     

The herpesvirus-Porphyromonas gingivalis-periodontitis axis

OBJECTIVES AND BACKGROUND: Members of the herpesvirus family have accumulated considerable support for a role in severe types of periodontitis. This study aimed to examine whether human cytomegalovirus (HCMV), Epstein-Barr virus type 1 (EBV-1) or herpes simplex virus (HSV) together with the major periodontopathic bacterium Porphyromonas gingivalis might interact in the pathogenesis of periodontal breakdown. METHODS: Sixteen subjects each contributed paper point samples from two progressing and two stable periodontitis lesions, as determined by ongoing loss of probing attachment. Polymerase chain reaction methodology was used to identify subgingival herpesviruses, P. gingivalis and other bacterial pathogens. Chi-squared tests and multivariate logistic regression were employed to identify statistical associations between herpesviruses, periodontopathic bacteria and clinical variables. RESULTS: HCMV and HSV were both significant predictors of the presence of subgingival P. gingivalis. In turn, P. gingivalis was positively associated with periodontitis active disease, probing attachment level, probing pocket depth, gingival bleeding upon probing and patient age. EBV-1 was not linked to P. gingivalis, although the virus was predictive of periodontitis active disease. The periodontitis disease risk associated with herpesvirus-P. gingivalis combinations depended on both site-specific and subject-specific factors. CONCLUSION: The present data of aggressive periodontitis implicate HCMV, HSV and P. gingivalis as either cofactors in its etiology or triggers of relapses. Further studies are needed to determine the spectrum of periodontopathogenicity of herpesviruses and effective management of these viruses in periodontal sites.     

Selective inhibition of Porphyromonas gingivalis growth by a factor Xa inhibitor, DX-9065a

BACKGROUND: Porphyromonas gingivalis is a causative bacterium of adult periodontitis. However, there is no drug specific for P. gingivalis and for its virulence factor. OBJECTIVES: The objective of this study was to examine the effects of a new selective inhibitor of activated factor X, DX-9065a, on growth of Porphyromonas gingivalis and other periodontopathic bacteria. METHODS: We incubated P. gingivalis and other periodontopathic bacteria in the presence or absence of DX-9065a and examined the effect of DX-9065a on bacterial growth and trypsin-like activity in its cultures. We also examined the effects of DX9065a on amidolytic activity of purified trypsin-like proteinases (gingipains RgpA and RgpB), from P. gingivalis and on trypsin-like activity in gingival crevicular fluids from patients with adult periodontitis. RESULTS: DX-9065a selectively inhibited the growth of P. gingivalis and Prevotella intermedia, and its effect on P. gingivalis was bactericidal. Trypsin-like proteinase activity was detected in P. gingivalis, and the activity was strongly inhibited by DX-9065a. DX-9065a even inhibited amidolytic activity of RgpA and RgpB from P. gingivalis. Furthermore, trypsin-like proteinase activity in gingival crevicular fluids was strongly inhibited by DX-9065a. CONCLUSIONS: DX-9065a inhibits P. gingivalis growth in part through to its ability to inhibit the trypsin-like proteinase activity in P. gingivalis and may be useful for a new drug for treatment of adult periodontitis.     

The effects of extracts from periodontopathic bacteria on human periodontal fibroblasts stimulated with mineralization supplements

Bacterial effects on in vitro mineralization of human periodontal fibroblasts (HPF) have not yet been examined in great detail. In our study, we investigated the effects of soluble extracts of the periodontopathic bacteria Porphyromonas gingivalis, Bacteroides forsythus and, Treponema denticola on cell proliferation, mineralization, as well as on osteoblastic markers present in HPF cultured in vitro, such as alkaline phosphatase (ALP) activity and collagen content. Periodontal fibroblasts stimulated by B-glycerophosphate, ascorbic acid and dexamethasone (BAD) or by dexamethasone and ascorbic acid (DA) were compared to unstimulated cells. During the cultivation period, the stimulation of HPF by combined dexamethasone and ascorbic acid (DA) had a strong inductive effect on proliferation, ALP activity and collagen formation. The extracts obtained from the periodontal pathogens had a suppressing effect on the proliferation rate of HPF. The extracts from P. gingivalis, B. forsythus and T. denticola caused a decrease in ALP activity within 24 h of application. While extracts obtained from P. gingivalis and B. forsythus induced a reduction in collagen content in BAD- and DA-stimulated HPF cells, T. denticola extracts led to an increase in collagen. Our data suggest that specific periodontopathic bacteria may suppress tissue regeneration in vivo not only by activating host defense mechanisms but also directly via a suppression of growth and differentiation of HPF and a reduction in the extracellular collagen matrix. For the process of pocket formation, not even the direct influence of viable bacteria seems to be necessary. Additionally, long-distance effects of bacteria harboured in periodontal pockets or in root canals may be of importance.     

Serum rheumatoid factor induced by intraperitoneal administration of periodontopathic bacterial lipopolysaccharide in mice

Serum rheumatoid factor (RF) level and peritoneal and splenic CD5⁺B (B-1) cells in mice were examined after intraperitoneal administration of purified lipopolysaccharides (LPS) from oral periodontopathic bacteria; Porphyromonas gingivalis, Actinobacillus actinomycetemcomitans, Fusobacterium nucleatum and Capnocytophaga ochracea. F. nucleatum and C. ochracea LPS induced higher levels of serum IgM- and IgG-RF, while P. gingivalis LPS showed the least induction. In addition, wet weights of spleen and serum IgM and IgG concentration were markedly increased in F. nucleatum LPS injected group. On the other hand, the proportion of CD5⁺ B cells to lymphocytes in the peritoneal cavity and spleen did not increase. The reason for this was not clear but conventional B cells (CD5⁺ B cells) might increase more rapidly with splenic enlargement than CD5⁺ B cells. These results suggested that RF induced by bacterial LPS may modulate immune responses against bacteria and plays an important role for defence and destruction of periodontal tissue.     

Induction of anti-thymocyte/T lymphocyte antibodies in mice injected with lipopolysaccharides from periodontopathic bacteria

We examined the levels of anti-thymocyte/T lymphocyte autoantibody (ATA) in the serum of mice injected intraperitoneally with lipopolysaccharides (LPS) from periodontopathic bacteria; Porphyromonas gingivalis, Actinobacillus actinomycetemcomitans, Fusobacterium nucleatum, Capnocytophaga ochracea, and non-oral Escherichia coli. All of the LPS induced IgM-ATA. Among these, LPS from C. ochracea induced the highest level of IgM-ATA, whereas that of P. gingivalis induced the lowest. The peritoneal T lymphocytes of mice injected with LPS were bound by IgM-ATA. Peritoneal B-1 (CD5+B) cells stimulated by each LPS produced much more IgM-ATA than splenic B-2 (CD5-B) cells, suggesting that B-1 cells might be responsible for the production of these antibodies. Serum of mice injected with C. ochracea and F. nucleatum LPS showed cytotoxicity against thymocytes in the presence of rabbit complements. Binding and cytotoxicity were confirmed by IgM purified from serum of the mice injected with C. ochracea LPS. Furthermore, serum of mice treated with C. ochracea, F. nucleatum or A. actinomycetemcomitans LPS inhibited the proliferation of thymocytes. However, purified IgM from the serum of mice treated with C. ochracea LPS failed to produce the same inhibition. Our results suggest that LPS from certain species of periodontopathic bacteria can induce IgM-ATA in the serum and these antibodies may modulate the local immune network in periodontal tissues.     

Hen egg-white lysozyme inhibits biological activities of lipopolysaccharides from periodontopathic bacteria

Lysozyme has a bactericidal activity for some strains of Gram-positive bacteria, by enzymatic cleavage of peptidoglycans that constitute the cell wall. Hen egg-white lysozyme (HEL) was tested in vitro for effects on biological activities of lipopolysaccharides from periodontopathic Gram-negative bacteria, Actinobacillus actinomycetemcomitans, Prevotella intermedia and Porphyromonas gingivalis . HEL inhibited a wide range of activities of these lipopolysaccharides, including activation of Limulus amoebocyte lysate, stimulation of human leukocytes to secrete tumour necrosis factor-alpha, polyclonal activation of mouse B cells, and promotion of osteoclastic differentiation in mouse bone marrow cultures. The anti-endotoxic activity of HEL may be worthy of being intended for periodontal therapy.     

An in vitro model to study bacterial invasion of periodontal tissues

In periodontal disease, the abilities of bacteria to adhere to and degrade in vivo basement membranes should be considered as two of the rate-limiting steps for the potential active or passive invasion of gingival connective tissues. To study these mechanisms in greater detail, we used the PF HR-9 basement-membrane-like matrix to establish an in vitro model of bacterial invasion and degradation. Three gram-negative anaerobic periodontopathic organisms, Bacteroides gingivalis, Fusobacterium nucleatum, and Actinobacillus actinomycetemcomitans, bound in considerably higher numbers to the HR-9 matrix than did 6 strains of gram-positive facultative organisms typically associated with periodontal health. In a further experiment with B. gingivalis, the organism rapidly degraded Type IV collagen, the major macromolecular component constituting the HR-9 matrix. Streptococcus mitis, the nonperiodontopathic bacterium tested, did not degrade this model matrix. This study provides evidence that B. gingivalis, a periodontopathic bacterium, is able to adhere to and degrade basement membranes, whereas nonperiodontopathic organisms appear not to share in these abilities.     

Identification of periodontopathic bacteria DNA from periodontitis-involved gingival tissue using a PCR method

The purpose of this study was to determine the presence of periodontopathic bacteria DNA in the granulation tissue of periodontally involved gingival tissue. Forty periodontitis patients were examined. Subgingival plaque and saliva samples were collected from the patients before surgery. Then, granulation tissue was also collected during surgery. Another 20 patients, who had more than 4 mm in probing pocket depth, were also examined as a control. They received only initial treatment without periodontal surgery. Plaque and saliva samples were also collected from them. Seven periodontopathic bacteria, Porphyromonas gingivalis, Bacteroides forsythus, Toreponema denticola, Campylobacter rectus, Actinobacillus actinomycetemcomitans, Prevotella intermedia, and Prevotella nigrescens, were detected by a PCR method from these samples of all patients. As clinical parameters, probing pocket depth, clinical attachment level, and bleeding on probing were recorded. One year after surgery, sampling of subgingival plaque and saliva, and measurement of clinical parameters were performed on the patients. The DNA of periodontopathic bacteria was detected in the granulation tissue of 29 out of 40 patients. P. gingivalis, B. forsythus, T. denticola, C. rectus, A. actinomycetemcomitans, P. intermedia and P. nigrescens were detected in 23, 11, 12, 9, 3, 2 and 1 subject(s) respectively. The sites with undetectable level of any periodontal pathogens at one year after surgery showed a clinical attachment gain of 2.44 mm, while those that showed detectable pathogen levels had only a 0.77 mm gain. Our results indicated that the removal of granulation tissue contributed to eliminating periodontal pathogens and to avoiding re-infection by those bacteria. Complete eradication of periodontopathic bacteria would be necessary for successful periodontal treatment.     

Cross-reactivity of GroEL antibodies with human heat shock protein 60 and quantification of pathogens in atherosclerosis

BACKGROUND/AIMS: Chronic infections such as those caused by Chlamydia pneumoniae and periodontopathic bacteria such as Porphyromonas gingivalis have been associated with atherosclerosis, possibly due to cross-reactivity of the immune response to bacterial GroEL with human heat shock protein (hHSP) 60. METHODS: We examined the cross-reactivity of anti-GroEL and anti-P. gingivalis antibodies with hHSP60 in atherosclerosis patients and quantified a panel of six pathogens in atheromas. RESULTS: After absorption of plasma samples with hHSP60, there were variable reductions in the levels of anti-GroEL and anti-P. gingivalis antibodies, suggesting that these antibodies cross-reacted with hHSP60. All of the artery specimens were positive for P. gingivalis. Fusobacterium nucleatum, Tannerella forsythia, C. pneumoniae, Helicobacter pylori, and Haemophilus influenzae were found in 84%, 48%, 28%, 4%, and 4% of arteries, respectively. The prevalence of the three periodontopathic microorganisms, P. gingivalis, F. nucleatum and T. forsythia, was significantly higher than that of the remaining three microorganisms. CONCLUSIONS: 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 be a possible mechanism for the association between atherosclerosis and periodontal infection.