IL-8 and TNF-alpha from peripheral neutrophils and acute-phase proteins in periodontitis

OBJECTIVE: In other studies, we have found deviant functions in peripheral neutrophils in periodontitis. The aim here was to study (1) the release of cytokines, IL-8 and TNFalpha, from neutrophils in 15 treated periodontitis patients and pair-matched controls as well as (2) the effects of cigarette smoking. MATERIAL AND METHODS: Cytokines released in the incubation medium from un-stimulated and Fcgamma-R-stimulated neutrophils and some acute-phase reactants were measured with ELISA. RESULTS: Non-smoking patients had trends for lower TNFalpha release compared to non-smoking controls, while corresponding trends were rather similar for Il-8. Smoking had a moderate but inconsistent effect on the release of both cytokines. However, in patients, the ratio between stimulated/un-stimulated release of Il-8 was significantly lowered by smoking (p<0.03). The parameters of inflammation in plasma differed only slightly between patients and controls, indicating that periodontal disease in a quiet phase has a negligible systemic effect with the possible exception for a higher IL-8 level. In contrast, smoking had significant systemic effect on the neutrophil count and IgG levels. CONCLUSIONS: Release of IL-8 and TNF-alpha from peripheral neutrophils and various parameters of inflammation in plasma seem to be affected more by cigarette smoking than periodontal disease.     

Juvenile and rapidly progressive periodontitis

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

快速进展性牙周炎患者外周血单核细胞趋化功能的研究

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

Immunodominant antigens of Porphyromonas gingivalis in patients with rapidly progressive periodontitis

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

Determination of serum antibody against Bacteroides gingivalis from rapidly progressive periodontitis and juvenile periodontitis patients

Bacteroides gingivitis (Bg) is one of the major pathogens associated with periodontitis. This is the first report in China on serum antibody level against Bg from patients with rapidly progressive periodontitis (RPP) and juvenile periodontitis (JP) using ELISA method. 21 RPP patients, 20 JP patients and 30 healthy subjects (H) were involved in this study. The results showed that the ratio of positive antibody response was 100% in RPP, 80% in JP and 30% in control group. The antibody response in both RPP and JP groups were significantly greater than that in healthy group (P less than 0.001). The difference between RPP and JP groups were also statistically significant (P less than 0.05).     

IL-1beta and TNF-alpha regulate IL-6-type cytokines in gingival fibroblasts

Interleukin-6 (IL-6)-type cytokines are pleiotropic molecules capable of stimulating bone resorption and expressed by numerous cell types. In the present study, we tested the hypothesis that gingival fibroblasts may exert local osteotropic effects through production of IL-6 and related cytokines. IL-6-type cytokine expression and regulation by IL-1beta and tumor necrosis factor-alpha (TNF-alpha) were studied in fibroblasts from the non-inflamed gingiva of healthy individuals. Constitutive mRNA expression of IL-6, IL-11, and leukemia inhibitory factor (LIF), but not of oncostatin M (OSM), was demonstrated, as was concentration-dependent stimulation of IL-6 and LIF mRNA and of protein by IL-1beta and TNF-alpha. IL-11 mRNA and protein were concentration-dependently stimulated by IL-1beta. The signaling pathway involved in IL-6 and LIF mRNA stimulation involved MAP kinases, but not NF-kappaB. The findings support the view that resident cells may influence the pathogenesis of periodontal disease through osteotropic IL-6-type cytokine production mediated by activation of MAP kinases. Abbreviations: IL-1alpha (interleukin-1alpha); IL-1beta (interleukin-1beta); IL-6 (interleukin-6); IL-11 (interleukin-11); LIF (leukemia inhibitory factor); OSM (oncostatin M); alpha(1)-coll. I (alpha(1)-collagen I); ALP (alkaline phosphatase); BMP-2 (bone morphogenetic protein-2); OC (osteocalcin); BSP (bone sialoprotein); TNFR I (tumor necrosis factor receptor I); TNFR II (tumor necrosis factor receptor II); IL-1R1 (interleukin-1 receptor 1); GAPDH (glyceraldehyde-3-phosphate dehydrogenase); RPL13A (ribosomal protein L13A); mRNA (messenger ribonucleic acid); cDNA (complementary deoxyribonucleic acid); PCR (polymerase chain-reaction); BCA (bicinchoninic acid); ELISA (enzyme-linked immunosorbent assay); alpha-MEM (alpha modification of Minimum Essential Medium); and FCS (fetal calf serum).     

Superoxide formation and chemiluminescence of peripheral polymorphonuclear leukocytes in rapidly progressive periodontitis patients

Previous studies have indicated that in certain types of chronic inflammatory periodontal diseases, polymorphonuclear leucocytes (PMN) functions are impaired. In view of the damage oxygen free radicals may cause to the periodontal tissues, the present study focussed on superoxide (SO) formation and luminol-dependent chemiluminescence (LDCL) by peripheral PMN cells in rapidly progressive periodontitis patients (RPP). PMN cell preparations were stimulated by either opsonized bacteria or phorbol myristate acetate (PMA). The results indicate that PMN cells from RPP patients, stimulated by opsonized bacteria, have significantly enhanced SO formation and LDCL response as compared to healthy subjects. The hyperactivity was cell-associated. In the presence of PMA, no significant differences were detected between the groups. The results suggest that PMN cells from RPP patients are functionally activated, and produce elevated levels of oxygen radicals. These oxygen radicals may play a role in the pathogenesis of RPP.     

Peripheral T lymphocyte subsets in rapidly progressive periodontitis

The peripheral blood lymphocyte subsets of 10 patients with rapidly progressive periodontitis were investigated with monoclonal antibody. In 4 patients, the T helper/T suppressor ratio was increased. 5 others had a slightly reduced T helper/T suppressor ratio as compared to control group. These findings may indicate a possible cellular immune response in the pathogenesis of rapidly progressive periodontitis.     

Psychosocial factors and adult onset rapidly progressive periodontitis

Abstract On the basis of clinical observations, some periodontologists have suggested an association between psychosocial factors such as depression, stress and anxiety, and adult onset rapidly progressive periodontitis (RPP). This study investigated more formally possible associations between a number of relevant psychosocial factors and RPP The significance of the psychosocial variables was assessed by comparing 3 groups: 50 patients with RPP, 50 patients with routine chronic adult periodontitis (RCAP). and 50 patients without significant periodontal destruction (controls). It was anticipated that the RPP group would show higher levels of psychosocial maladjustment than the RCAP and control groups. A between-subjects multivariate analysis of covariance indicated that the combined psychosocial variables were significantly related to the periodontal diagnosis. 2 psychosocial factors, depression and loneliness, were significant in distinguishing between groups. The RPP group presented significantly increased depression and loneliness compared to the RCAP and control groups. Future research is indicated to further clarify the significance of these psychosocial differences in relation to the onset and progression of RPP.     

Reaction of human sera from juvenile periodontitis, rapidly progressive periodontitis, and adult periodontitis patients with selected periodontopathogens

The levels of serum antibody reactive to selected periodontopathogens were determined in 182 clinically characterized patients: 35 healthy control, 50 juvenile periodontitis, 42 adult periodontitis and 55 rapidly progressive periodontitis. Reactive antibody levels were determined using an enzyme-linked immunosorbent assay with whole cell preparations of Bacteroides gingivalis, Capnocytophaga (Bacteroides) ochraceus, Fusobacterium nucleatum and Actinobacillus actinomycetemcomitans (Y-4) serving as antigens. Increased reactivity to B. gingivalis and F. nucleatum was observed in all three disease groups studied while antibody reactive to A. actinomycetemcomitans was increased in juvenile and rapidly progressive periodontitis. Antibody levels reactive to C. ochraceus in healthy subjects did not differ from those observed in any disease patient groups. Possible implications in the etiology and progression of the diseases coupled with environmental changes which occur in the econiche of the periodontal pocket are described.     

快速进展性牙周炎患者龈沟液中的弹性蛋白酶活性

目的探讨中性多形核白细胞与快速进展性牙周炎的关系。方法检测２例快速进展性牙周炎患者共１０２个位点（其中２８个位点进行治疗前后的对比）的龈沟液弹性蛋白酶活性,将人多形核白细胞弹性蛋白酶特异底物———Ｓ２４８４与龈沟液反应,测吸光度值,以反映龈沟液弹性蛋白酶活性。结果快速进展性牙周炎龈沟液中的弹性蛋白酶活性［（０．６３±０．３８）Ａｂｓ／位点］明显高于健康对照组［（０．０７±０．０５）Ａｂｓ／位点］,差异有显著性;弹性蛋白酶活性的高低与龈沟液体积、探诊深度、附着丧失和出血指数呈正相关关系;治疗后龈沟液体积和各临床指数显著降低,弹性蛋白酶活性也从治疗前的（０．７３±０．３６）Ａｂｓ／位点下降为（０．１±０．１７）Ａｂｓ／位点,差异有显著性。结论快速进展性牙周炎患者的中性多形核白细胞并不是趋化反应不足,而是过度浸润与释放溶酶体酶,起协同致炎作用。     

Immunohistological and morphometric analysis of inflammatory cells in rapidly progressive periodontitis and adult periodontitis

The purpose of this study was to localize, characterize, and quantify in situ the inflammatory cells in the gingival connective tissue prior and subsequent to the initial therapy of ten patients with rapidly progressive periodontitis (RPP) and five patients with adult periodontitis (AP). Using immunohistological techniques, the amount of T lymphocytes, alphabeta-T lymphocytes, gammadelta-T lymphocytes, B lymphocytes, and plasma cells was determined at the beginning of the periodontal therapy (baseline) and at the time of periodontal surgery. Furthermore, the distribution of collagen types I, III, V, and VI was investigated using transmission electron microscopy. At baseline, patients with RPP revealed much higher numbers of inflammatory cells than patients with AP. During initial therapy of patients with RPP, the amount of T cells, alphabeta-T cells, and gammadelta-T cells was reduced significantly (P<0.05). Biopsies of patients with AP revealed a statistically significant reduction of all cell types, except alphabeta-T cells and gammadelta-T cells in the deep connective tissue. The transmission electron microscopy of biopsies from patients with RPP and AP with severe inflammation taken at baseline revealed that collagen types I and III were destroyed nearly completely in areas with leukocyte infiltration, whereas collagen types V and VI revealed a more pronounced labeling reaction. The results revealed that, during initial therapy, the amount of inflammatory cells was reduced significantly more in biopsies of patients with AP than in patients with RPP. At baseline, the inflamed gingival tissue consists mainly of collagen types V and VI in areas with infiltrates of inflammatory cells.     

Phagocytosis and adhesiveness of peripheral blood polymorphonuclear leukocytes in patients with rapidly progressive periodontitis

Phagocytosis and adhesiveness of peripheral blood polymorphonuclear leukocytes were studied in patients diagnosed with generalized rapidly progressive (form A) periodontitis. All patients belonged to a single family and were first degree relatives. In the patients' leukocytes there was a statistically significant reduction in phagocytic index and adhesiveness, in comparison with leukocytes from controls. However, adhesiveness approached control values in the presence of autologous serum, thus implying that serologic factors may enhance this function in patients with this type of periodontitis.     

TNF-alpha and IL-4 levels in generalized aggressive periodontitis subjects

Objectives:  The aim of this study was to evaluate tumor necrosis factor (TNF)-α and interleukin (IL)-4 levels in healthy sites and sites exhibiting signs of moderate and advanced generalized aggressive periodontitis (GAgP) in the same subject.
Methods:  The following sites were selected for crevicular fluid sampling in the same AgP subject ( n  = 14): Healthy sites (HS): no marginal bleeding or bleeding on probing (BOP) and probing depth (PD) ≤ 3 mm; Moderate sites (MS): BOP and PD between 4 and 6 mm; Advanced sites (AS): BOP and PD ≥ 7 mm. One site from periodontally healthy subjects ( n  = 13) was sampled for use as a control. TNF-α and IL-4 levels were measured using ELISA.
Results:  The total amount of TNF-α was lower for control sites, while there were no differences among healthy and diseased sites from GAgP subjects ( P  < 0.05). The concentration of TNF-α was higher in HS, in relation to the other sites ( P  < 0.05). There were no significant differences among the groups regarding total amounts of IL-4 ( P  > 0.05), while IL-4 concentration was significantly higher in control sites, when compared with sites from GAgP subjects ( P  < 0.05).
Conclusion:  In conclusion, high levels of TNF-α and low levels of IL-4 were observed in both healthy and diseased sites within the same generalized AgP individuals.     

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

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

快速进展性牙周炎患者的白细胞趋化功能

目的了解我国快速进展性牙周炎患者的外周血中性多形核白细胞是否存在趋化功能异常。方法选择细菌源性的趋化肽和宿主源性的趋化因子（白细胞介素８）作为趋化剂，采用微孔滤膜法（改良的ＢｏｙｄｅｎＣｈａｍｂｅｒ法）检测１６例快速进展性牙周炎患者的外周血中性多形核白细胞的趋化功能，并与１４名年龄相当的健康个体进行对照。结果快速进展性牙周炎患者的外周血中性多形核白细胞对细菌源性的趋化肽和白细胞介素８的趋化反应与健康组差异无显著性。结论中性多形核白细胞趋化功能的异常可能并不是快速进展性牙周炎的主要易感因素。     

Treatment considerations in rapidly progressive periodontitis: a case report

This article presents a concept for the control of periodontal pathogens in early-onset periodontitis and demonstrates the reparative potential of periodontal tissue when the infection is under control. The patient discussed here was diagnosed with rapidly progressive periodontitis. We were able to reduce the bacterial mass with scaling and root planing and, in conjunction with systemic antibiotics, return the microbial profile to the normal range. Microbiological testing was used to monitor the microbiota and to adjust antimicrobial treatment. Improvements in probing depths and attachment levels were monitored for more than 1 year. Tissue response to this treatment made surgical intervention unnecessary. Although not essential, orthodontic treatment enhanced cleansability and improved esthetics.     

The ATPase content of the blood neutrophils and lymphocytes in patients with chronic periodontitis]

Chronic periodontitis is associated with reduced immunologic reactivity of the body, that may present as disturbed status of the lymphocytes and neutrophils due to reduced activity of adenosine triphosphatases. Drug therapy of periodontitis, physiotherapy, oral hygiene, sodium chloride reducing diets are conductive to improvement of the clinical status of the periodontium and to recovery of ATPase activities.