The influence of interleukin gene polymorphism on expression of interleukin-1beta and tumor necrosis factor-alpha in periodontal tissue and gingival crevicular fluid.

BACKGROUND: A specific composite genotype of the polymorphic interleukin-1 (IL-1) gene cluster has recently been associated with severe periodontitis. One polymorphism of the composite periodontitis-associated genotype (PAG) has been functionally linked with expression of high levels of IL-1. The purpose of this study was to test whether gingival crevicular fluid (GCF) levels of IL-1beta and tumor necrosis factor-alpha (TNFalpha), and gingival tissue levels of IL-1alpha, IL-1beta, and TNFalpha correlate with PAG, and to examine the effect of conservative periodontal therapy on these levels. METHODS: Twenty-two adults with moderate to advanced periodontal disease were enrolled. Polymerase chain reaction amplification and restriction enzymes were used to identify specific polymorphisms from peripheral blood samples. GCF samples were collected at baseline and 3 weeks following conservative treatment and analyzed by ELISA for IL-1beta and TNFalpha. An interproximal gingival biopsy was collected at baseline and follow-up and analyzed for IL-1alpha, IL-1beta, and TNFalpha by ELISA. RESULTS: The genotyping identified 7 as PAG(+) and 15 as PAG(-). The 2 groups were comparable in terms of existing periodontitis and age. In shallow sites (<4 mm), total IL-1beta in GCF was 2.5 times higher for PAG(+) patients prior to treatment (P=0.03), and 2.2 times higher after treatment (P=0.04), while differences were less apparent in deeper sites. Following treatment, a reduction in IL-1beta concentration in GCF was seen for PAG(-) but not for PAG(+) patients. While not statistically significant, a trend was observed in mean tissue levels of IL-1beta which were 3.6 times higher in PAG(+) versus PAG(-) patients (P=0.09). CONCLUSIONS: These data suggest that PAG(+) patients may demonstrate phenotypic differences as indicated by elevated levels of IL-1beta in GCF.     

Soluble antagonists to interleukin-1 (IL-1) and tumor necrosis factor (TNF) inhibits loss of tissue attachment in experimental periodontitis

BACKGROUND, AIMS: Periodontal disease is a significant cause of tooth loss among adults and is characterized by the alteration and permanent destruction of the deeper periodontal tissues. Although the presence of pathologic microbes is required to trigger this process, the amplification and progression of the diseased state is believed to rely heavily on the production of host mediators in response to bacteria or their metabolic products. The inflammatory response is effective in preventing large-scale colonization of the gingival tissues by bacteria that lie in close proximity to the tooth surface or within the gingival sulcus. It has been postulated that the host-response in some individuals may lead to an over-reaction to invading oral pathogens resulting in the destruction of periodontal tissues. METHODS: Several host-derived mediators are believed to contribute to this response. Two agents considered to be essential in periodontal destruction are interleukin-1 (IL-1) and tumor necrosis factor (TNF). We investigated the role of IL-1 and TNF in the loss of connective tissue attachment in a Macaca fascicularis primate model of experimental periodontitis. Silk ligatures impregnated with the periodontal pathogen, Porphyromonas gingivalis were wrapped around the posterior teeth and the activity of IL-1 and TNF were inhibited by soluble receptors to these proinflammatory cytokines via local injection into interdental papillae. RESULTS: Histomorphometric analysis indicates that IL-1 and TNF antagonists significantly reduced the loss of connective tissue attachment by approximately 51% and the loss of alveolar bone height by almost 91%, both of which were statistically significant. CONCLUSION: This investigation demonstrates that the loss of connective tissue attachment and progression of periodontal disease can be retarded by antagonists to specific host mediators such as IL-1 and TNF and may provide a potential treatment modality to combat the disease process.     

Lipopolysaccharide from Actinobacillus actinomycetemcomitans stimulates macrophages to produce interleukin-1 and tumor necrosis factor mRNA and protein

Actinobacillus actinomycetemcomitans is associated with periodontal disease in children and adults. We report that low concentrations of lipopolysaccharide (LPS) from A. actinomycetemcomitans stimulated human macrophages to increase dramatically their accumulation of mRNA coding for interleukin-1 alpha (IL-1 alpha), IL-1 beta as well as tumor necrosis factor (TNF). Protein levels of IL-1 and TNF alpha also increased. Levels of these mRNAs increased by 4-5 fold as compared with unstimulated macrophages when these cells were cultured with as little as 2 ng/ml LPS from A. actinomycetemcomitans. Polymyxin binds and blocks the action of LPS; polymyxin inhibited the ability of LPS from A. actinomycetemcomitans to increase levels of IL-1 beta mRNA. The LPS of A. actinomycetemcomitans stimulated increased levels of IL-1 beta mRNA in the presence of cycloheximide, showing that stimulation by this LPS did not require new synthesis of protein. Furthermore, dexamethasone inhibited the ability of LPS from A. actinomycetemcomitans to stimulate the accumulation of mRNA coding for IL-1 beta. A. actinomycetemcomitans is an invasive microorganism of the gingiva; high intragingival numbers correlate with sites undergoing local destruction of the periodontium. IL-1 alpha, IL-1 beta, and TNF are potent monokines that mediate inflammation and resorption of bone. Out studies suggest that macrophages migrating to these gingival sites of A. actinomycetemcomitans infection will be stimulated by LPS of A. actinomycetemcomitans to produce IL-1 alpha, IL-1 beta and TNF. These cytokines will mediate gingival inflammation and stimulate resorption of alveolar bone.     

Actinobacillus actinomycetemcomitans and Bacteroides gingivalis activate human peripheral monocytes to produce interleukin-1 and tumor necrosis factor

The effects of gram-negative bacteria clearly associated with juvenile and adult periodontitis on monokine production were assessed using standard in vitro assay techniques. Actinobacillus actinomycetemcomitans and Bacteroides gingivalis were able to activate human peripheral blood monocytes to produce significant amounts of interleukin-1 (IL-1) and tumor necrosis factor (TNF). These monokines are known to induce osteoclastic bone resorption. An oral gram-positive organism, Staphylococcus epidermidis, was able to induce only modest amounts of IL-1 and TNF, slightly above unstimulated monocyte levels.     

Localization of interleukin-1 beta in human periodontal tissue

Interleukin-1 beta (IL-1 beta) is the predominant form of IL-1 produced by macrophages. IL-1 beta possesses numerous and diverse biological activities. Several of these activities, including fibroblast proliferation, potentiation of the immune response, and stimulation of bone resorption may be of relevance to the pathogenesis of periodontal disease. This study was designed to examine the presence of IL-1 beta in human periodontal tissue. An antiserum directed against the N-terminal segment (117-131) of human IL-1 beta was used to detect IL-1 beta using immunofluorescent staining techniques. IL-1 beta positive staining cells were observed in both normal and diseased tissue and were limited to the lamina propria. Brightly staining cells were increased by almost 3-fold in periodontally diseased tissue when compared to normal tissue. Low intensity staining cells were equally distributed in the normal and diseased specimens. We propose that IL-1 beta and IL-1 beta produced by cells in periodontal tissues may be related to the pathological processes associated with periodontal disease.     

盐酸小檗碱对大鼠牙周组织中白细胞介素-1β和肿瘤坏死因子-α表达的影响

目的探讨盐酸小檗碱对大鼠实验性牙周炎模型牙周组织中白细胞介素- 1β（IL- 1β）和肿瘤坏死因子- α（TNF- α）表达的影响。方法在成功建立大鼠实验性牙周炎模型的基础上，实验动物在分别灌服盐酸小檗碱后第1、2、3、4周处死，取牙周组织标本作切片，行常规苏木精- 伊红染色，观察牙周组织病理变化状况，并采用免疫组织化学SABC法检测IL- 1β、TNF- α水平变化。结果牙周炎组牙周组织中IL- 1β、TNF- α表达明显高于正常组（P<0.05）；各时间段牙周炎治疗组牙周组织中IL- 1β、TNF- α表达明显低于牙周炎组（P<0.05），但牙周炎治疗组各时间段之间IL- 1β、TNF- α表达无显著差异。结论大鼠实验性牙周炎模型牙周组织中TNF- α、IL- 1β表达水平较正常对照组明显升高，应用盐酸小檗碱能抑制实验性牙周炎模型牙周组织中TNF- α、IL- 1β的表达。     

Effect of soluble receptors to interleukin-1 and tumor necrosis factor alpha on experimentally induced root resorption in rats

OBJECTIVE: In this study, the role of the inflammatory cytokines interleukin-1 (IL-1) and tumor necrosis factor alpha (TNFalpha) in the course of mechanically induced root resorption was investigated. METHODS: Mechanical induction of root resorption was performed on the upper left first molars in 18 male Wistar rats according to the method of Nakane and Kameyama. Starting on day minus 1, six animals received daily intraperitoneal injections of 2 ml of 1 micro g/ml soluble receptors to IL-1 (sIL-1RII) and another six animals were administered the same dose of soluble receptors to TNFalpha (sTNFalpha-RI). Six animals served as a control. On d 7 the left maxillae were prepared for histological and morphometric analysis of the extent of the root resorption that had developed. RESULTS: The qualitative and quantitative results demonstrated that in both receptor groups the amount of root resorption was significantly reduced. Especially following systemic application of sTNFalpha-RI, root resorption was nearly completely prevented. CONCLUSIONS: Our results indicate that IL-1 and more particularly TNFalpha are important for the induction and the further process of mechanically induced root resorption in the rat.     

The interleukin-1 genotype as a severity factor in adult periodontal disease

Abstract Although specific bacteria, dental plaque, and age are associated with periodontal disease, there are currently no reliable predictors of periodontitis severity. Studies in twins have suggested a genetic contribution to the pathogenesis of periodontitis, but previous attempts to identify genetic markers have been unsuccessful. The pro-inflammatory cytokines interleukin-1 (IL-1) and tumor necrosis factor alpha (TNFα) are key regulators of the host responses to microbial infection. IL-1 is also a major modulator of extracellular matrix catabolism and bone resorption. We report a specific genotype of the polymorphic IL-1 gene cluster that was associated with severity of periodontitis in non-smokers, and distinguished individuals with severe periodontitis from those with mild disease (odds ratio 18.9 for ages 40–60 years). Functionally, the specific periodontitis-associated IL-1 genotype comprises a variant in the IL-1B gene that is associated with high levels of IL-1 production. In smokers severe disease was not correlated with genotype. In this study, 86.0% of the severe periodontitis patients were accounted for by either smoking or the IL-1 genotype. This study demonstrates that specific genetic markers, that have been associated with increased IL-1 production, are a strong indicator of susceptibility to severe periodontitis in adults.     

可溶性白细胞介素-1和肿瘤坏死因子受体对大鼠正畸牙移动影响的研究

目的研究局部注射重组人类可溶性受体对大鼠正畸牙移动进程的影响。方法选取64只雄性SD大鼠,从加力第1天起在大鼠牙齿局部分别注射重组人类可溶性白细胞介素- 1受体Ⅱ（sIL- 1- RⅡ）、可溶性肿瘤坏死因子受体Ⅰ（sTNF- RⅠ）、两者混合物,测量磨牙移动距离,用苏木精- 伊红（HE）染色观察被移动磨牙牙周组织形态学变化,并应用抗酒石酸盐酸性磷酸酶（TRAP）组织化学染色对破骨细胞和破牙骨质细胞的数量及分布的时相性改变进行分析。结果各受体组大鼠磨牙牙周组织形态学时相性变化与对照组相似,但各时间点牙槽骨吸收程度均较轻微,多数大鼠牙根表面很少或几乎没有明显牙骨质吸收迹象。与对照组相比,在第14天所有受体注射组牙槽骨及牙根表面TRAP染色阳性细胞降低了约50%（P<0.05）,但受体组组间差异无统计学意义（P>0.05）。结论在大鼠正畸牙局部注射重组人类可溶性白细胞介素- 1和肿瘤坏死因子受体,可以减少牙齿移动距离并减少牙根吸收。     

辅助性T细胞亚群与牙周炎的免疫损伤机制

牙周炎症组织破坏与机体针对致病菌及其毒性产物的免疫反应方式有关,辅助性T细胞（Th）亚群通过分泌不同的细胞因子发挥免疫调节功能。较早发现的Th1和Th2分别介导细胞免疫和体液免疫,二者相互抑制保持平衡,牙周炎症的间断性和进展性与Th1/Th2主导的免疫反应的平衡破坏有关。在牙周炎早期和静止期,表现为Th1反应类型;在牙周炎晚期或活动期,则表现为Th2反应类型。Th17具有重要的促进牙周慢性炎症及免疫损伤形成的作用,而调节性T细胞（Treg）则具有炎症抑制作用,而且Th17/Treg在功能和分化上也具有与Th1/Th2类似的相互调节相互抑制的平衡关系。Th17/Treg和Th1/Th2在局部形成复杂的细胞因子网络,是目前进一步研究牙周炎的免疫病理学机制的热点。     

��ϸ������-1���������ܲ��Ĺ�ϵ

白细胞介素-1(interleukin-1,IL-1)是人体内非常活跃的促炎细胞因子,可刺激内皮细胞和白细胞释放一系列炎性介质,由此引发局部组织或全身的的炎症.牙周病是发生在牙龈、牙周膜和牙槽骨等牙齿支持组织的一种慢性破坏性疾病,其病理机制与牙周局部炎症有密切的联系.本文就IL-1的分化、功能及其与牙周病的关系做一综述.     

肿瘤坏死因子对培养人牙周膜成纤维细胞c-fos表达的诱导作用

目的：深入探索肿瘤坏死因子对人牙周膜成纤维细胞的生物学功能影响的机制。方法：将５０００Ｕ／ｍｌ肿瘤坏死因子作用于培养至第三代的人牙周膜细胞３ｈ后，进行ＦＯＳ免疫组化反应。结果：几乎所有的ＰＤＬＦｓ核均染为棕黄色，为ＦＯＳ免疫组化反应阳性，对照组细胞核未见着色。结论：由于ｃ－ｆｏｓ的表达可以提示核内基因的进一步改变，因而从更深的层次上说明肿瘤坏死因子对牙周膜成纤维细胞的生物学功能的影响     

Salivary interleukin-6 and tumor necrosis factor alpha in patients with drug-induced xerostomia

It is well known that cytokines are involved in the homeostasis of oral cavity and that altered levels of either serum and/or salivary cytokines have been found in certain oral/systemic diseases. So far, cytokines in connection with xerostomia have been investigated in patients with Sjögren's syndrome. We wanted to find out whether drugs themselves influence salivary glands, which would result in altered cytokine level or whether xerostomia itself of different causes leads to the changes in salivary cytokine levels. Therefore, the aim of this study was to evaluate levels of salivary interleukin‐6 (IL‐6) and tumor necrosis factor (TNF)‐α in 30 patients with drug‐induced xerostomia, age range 29–84 and mean 63.9 years. Control group consisted of 30 healthy participants, age range 30–82 years and mean age 65.2 years. Enzyme‐linked immunoassay was performed on commercially available kits. Statistical analysis was performed by use of Student's test. No significant differences in salivary IL‐6 and TNF‐α between patients with drug‐induced xerostomia when compared with the healthy controls were found (P < 0.05). We might conclude that drugs do not induce damage to the salivary glands which could be seen in altered salivary IL‐6 and TNF‐α levels and that xerostomia itself, induced by drugs does not alter levels of the investigated salivary cytokines.     

Analysis of tumor necrosis factor-alpha, interleukin-6, interleukin-1beta, soluble tumor necrosis factor receptors I and II, interleukin-6 soluble receptor, interleukin-1 soluble receptor type II, interleukin-1 receptor antagonist, and protein in the synovial fluid of patients with temporomandibular joint disorders

OBJECTIVE: To measure the levels of various cytokines, cytokine receptors, and cytokine antagonists in the synovial fluid of patients with temporomandibular disorders (TMD) and to determine the correlations among these expression levels. STUDY DESIGN: Synovial fluid was obtained from 55 patients with TMD and from 5 asymptomatic healthy volunteers as controls. The concentrations of tumor necrosis factor (TNF)-alpha, interleukin (IL)-6, IL-1beta, soluble tumor necrosis factor receptors I and II (sTNFR-I and sTNFR-II), IL-6 soluble receptor (IL-6sR), IL-1 soluble receptor type II, and IL-1 receptor antagonist were determined by enzyme-linked immunosorbent assay. RESULTS: The concentrations of TNF-alpha, IL-6, IL-1beta, sTNFR-I, and sTNFR-II were significantly higher in the synovial fluid of patients than in controls (P < .05). TNF-alpha level was positively correlated with those of IL-6, sTNFR-I, and sTNFR-II. In particular, there was a highly significant positive correlation between sTNFR-I and sTNFR-II. CONCLUSION: TNF and sTNFRs in the synovial fluid of patients with TMD may be important in the pathogenesis of these disorders.     

Investigation of functional gene polymorphisms interleukin-1β, interleukin-6, interleukin-10 and tumor necrosis factor in individuals with oral lichen planus

Oral lichen planus (OLP) is a chronic inflammatory oral mucosal disease. There are some studies in the literature demonstrating association between cytokines genes polymorphisms and susceptibility to develop some immune-mediate conditions. The purpose of this study was to investigate cytokine gene polymorphisms in a sample of Brazilian patients with OLP. Fifty-three patients with OLP (mean age = 43.1 years; range 20-68 years) and 53 healthy volunteers (mean age = 42.9 years; range 21-67) were genotyped for IL-1beta +3954 (C/T), IL-6-174 (G/C), IL-10-1082 (G/A) and TNFA-308 (G/A) gene polymorphisms. Statistical analysis was based on the use of logistic regression (P-values below 0.05 were considered as significant). IL-6 and TNFA homozygous genotypes were significantly more often detected in OLP patients. These genotypes were associated with an increased risk of OLP development (OR 6.89 and 13.04, respectively). IL-1beta and IL-10 gene polymorphisms were not related to OLP development. Our findings clearly demonstrate an association between inheritance of IL-6 and TNFA gene polymorphisms and OLP occurrence, thus giving additional support for genetic basis of this disease.