Objective: To characterize the salivary microbiota of patients with aggressive periodontitis, patients with chronica periodontitis and orally healthy individuals. Methods: A total of 81 unstimulated saliva samples from aggressive periodontitis patients (n = 31), chronic periodontitis patients (n = 25), and orally healthy controls (n = 25) were examined. The V1-V3 region of the 16S rDNA gene was sequenced with Illumina® MiSeq TM , and sequences were annotated to the expanded Human Oral Microbiome Database (eHOMD). Results: A mean percentage of 97.6 (range: 89.8-99.7) of sequences could be identified at species level. Seven bacterial species, including Porphyromonas gingivalis, were identified with significantly higher relative abundance in saliva from aggressive periodontitis patients than in saliva from orally healthy controls. Salivary abundance of P. gingivalis could discriminate aggressive (AUC: 0.80, p = 0.0001) and chronic periodontitis (AUC: 0.72, p = 0.006) from healthy controls. Likewise, salivary presence of P. gingivalis was significantly associated with aggressive (p < 0.0001, RR: 8.1 (95% CI 2.1-31.2)) and chronic periodontitis (p = 0.002, RR: 6.5 (95% CI: 1.6-25.9)). Conclusion: Salivary presence and relative abundance of P. gingivalis associate with aggressive and chronic periodontitis, but do not discriminate between aggressive and chronic periodontitis.
Background:The facultative bacterium Aggregatibacter actinomycetemcomitans (Aa) is strongly associated with periodontitis and is occasionally found in periodontally healthy subjects. We aimed to determine the prevalence of salivary Aa among patients with either periodontitis Grade B (periodontitis-B) or Grade C (periodontitis-C), periodontally healthy controls (HCs), and to determine if systemic antibodies against Aa or its virulence factor leukotoxin A (LtxA) may serve as biomarkers that reveal the oral presence of the bacterium and discriminate subjects with periodontitis-C, periodontitis-B, or no periodontitis from each other.Methods: Serum and unstimulated saliva samples were collected from patients with periodontitis-C (n = 27), patients with periodontitis-B (n = 34), and HCs (n = 28). Serum level of immunoglobulin G antibodies to fragmented whole Aa and to LtxA were quantified using a bead-based assay. Aa was identified in saliva using quantitative polymerase chain reaction (qPCR). All analyses were adjusted for age, sex, and current smoking status.Results: Aa was present in saliva from 11% of HCs, in 32% of patients with periodontitis-B (P = 0.04 versus HCs), and in 37% of patients with periodontitis-C (P = 0.02 versus HCs). Serum antibodies to fragments of Aa associated significantly with periodontitis-C (P = 0.03), while serum anti-LtxA antibodies associated with both periodontitis-B and periodontitis-C (P = 0.002 and P = 9×10 −4 , respectively). Moreover, a significant association between serum anti-LtxA antibodies and Aa count in saliva was observed (P = 0.001). On the basis of serum anti-LtxA antibody levels, patients with periodontitis could be discriminated from HCs (AUC = 0.74 in ROC curve-analysis, P = 0.0003), and carriers of Aa could be discriminated from non-carriers (AUC = 0.78, P <0.0001). Conclusions: Aa is highly prevalent in saliva of patients with periodontitis-B or periodontitis-C. Systemic immunoglobulin G antibodies against LtxA distinguish patients with periodontitis, regardless of grade, from HCs, while their quantity reflects the concurrent bacterial burden in the oral cavity.
Background Periodontitis (PD) is classified by Grades A through C according to the risk of further progression, PD Grade C (PD‐C) being the most severe progressing form. It is a matter of controversy, whether the disease activity observed in PD‐C is due to impaired immune reactivity toward bacteria embedded in biofilms or a hyper‐reactive immune response causing tissue damage as a bystander phenomenon. Little is known about the role of complement in this respect. Methods Plasma and unstimulated saliva samples were collected from patients with PD‐B (n = 34) or ‐C (n = 27) and healthy controls (HCs) (n = 28). Salivary and plasma levels of total C3, C3c, and C3dg were quantified using sandwich enzyme‐linked immunosorbent assay (ELISA). Results Salivary levels of total C3 and C3dg were elevated in PD‐B and PD‐C patients compared to HCs (both P < 0.05), while the levels of C3c were elevated in PD‐C compared to HCs. Plasma levels of C3c were higher in PD‐B patients than in HCs (P < 0.05). Conclusion PD‐B and PD‐C patients show increased complement activation compared to HCs, but no difference was found between the two disease grades. PD‐B, but not PD‐C, is associated with increased systemic complement activation as assessed by C3c in plasma.
BackgroundCytokine‐producing B cells play a well‐established role in modifying immune responses in chronic inflammatory diseases. We characterized B‐cell cytokine responses against periodontitis‐associated bacteria in patients with periodontitis.MethodsBlood and saliva samples were collected from patients with periodontitis grade B (N = 31) or grade C (N = 25), and 25 healthy controls (HCs). Mononuclear cells were stimulated with Porphyromonas gingivalis, Fusobacterium nucleatum, Staphylococcus epidermidis, or Cutibacterium acnes, and B‐cell production of tumor necrosis factor (TNF)‐α, interleukin (IL)‐6, interferon (IFN)‐γ, IL‐10 and transforming growth factor (TGF)‐β by B cells was assessed by flow cytometry.ResultsHCs had higher baseline frequencies of B cells producing IFN‐γ or TNF‐α than grade B patients, but only B cells from grade B patients showed significant differentiation into IFN‐γ‐, TNF‐α‐, TGF‐β‐, or IL‐10‐producing cells after challenge with P. gingivalis and into IFN‐γ‐, TGF‐β‐, or IL‐10‐producing cells after challenge F. nucleatum. Notably, the baseline frequency of IL‐10‐producing B cells from grade C patients correlated inversely with clinical attachment loss (AL). The major proportion of the IFN‐γ‐ and TGF‐β‐producing B cells were CD27+ memory cells, while the IL‐10‒producing B cells were mainly CD27−CD5−.ConclusionsB cells from grade B patients, particularly those harboring P. gingivalis, showed proinflammatory B‐cell responses to P. gingivalis. Moreover, the baseline frequency of IL‐10‐producing B cells in the grade C group correlated inversely with AL, suggesting a diminished immunoregulatory capacity of IL‐10‒producing B cells in these patients.
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