Enhanced interleukin-8 production in THP-1 human monocytic cells by lipopolysaccharide from oral microorganisms and granulocyte-macrophage colony-stimulating factor

Granulocyte-macrophage colony-stimulating factor (GM-CSF) has been used to assist in bone marrow recovery during cancer chemotherapy. Interleukin-8 (IL-8) plays an important role in macrophage mediated inflammatory processes including exacerbation of periodontal diseases, one of the most common complications in GM-CSF receiving cancer patients. The effect of GM-CSF supplementation on IL-8 production was investigated in a human monocyte cell line THP-1, stimulated with lipopolysaccharide extracted from two oral microorganisms, Porphyromonas gingivalis and Fusobacterium nucleatum. Resting THP-1 cells were treated with lipopolysaccharide (1 microgram/ml) of P. gingivalis or F. nucleatum and/or GM-CSF (50 IU/ml) for varying time periods. The production of IL-8 in THP-1 cells was measured by a solid-phase enzyme-linked immunosorbent assay (ELISA). A very low level of the cytokine IL-8 was produced constitutive in THP-1 cells. Starting from 8 h of treatment and afterwards GM-CSF alone significantly increased IL-8 production in THP-1 cells. Lipopolysaccharide (1 microgram/ml) extracts from either F. nucleatum or P. gingivalis amplified IL-8 production 500-800 times in comparison to resting THP-1 cells. When lipopolysaccharide of F. nucleatum or P. gingivalis was supplemented with 50 IU/ml of GM-CSF, there was a statistically significant enhanced production of IL-8 by THP-1 cells after 1 day to 7 days of treatment as compared with lipopolysaccharide treatment alone. GM-CSF (50 IU/ml) also significantly increased IL-8 production from 2-7 days of treatment of THP-1 cells when supplemented with a positive control, phorbol-12-myristate-13 acetate (PMA), as compared to PMA treatment alone. These investigations using the in vitro THP-1 human monocyte cell model indicate that there may be an increase in the response on a cellular level to oral endotoxin following GM-CSF therapy as evidenced by enhanced production of the tissue-reactive inflammatory cytokine, IL-8.     

Effects of the oral spirochete Treponema denticola on interleukin‐8 expression from epithelial cells

This communication demonstrates that the interaction of the oral spirochete Treponema denticola 35405 with KB epithelial cells does not lead to the induction of interleukin‐8 production as occurs with a variety of other bacteria. Utilizing the dentilisin protease mutant K1 of T. denticola, this property was demonstrated to be primarily a function of the expression of the protease by strain 35405.     

定向敲除牙龈卟啉单胞菌菌毛蛋白FimA的重组质粒构建

目的 构建定向敲除牙龈卟啉单胞菌(Porphyromonas gingivalis,Pg)菌毛蛋白FimA基因的质粒pPHU281_A_Spec_B,用于后续构建菌毛蛋白FimA缺陷型Pg,为研究FimA的功能奠定分子基础.方法 厌氧培养PgATCC33277,提取基因组DNA,聚合酶链反应扩增PgFimA基因一定长度的上游A片段及下游B片段,并在扩增片段两端添加合适的酶切位点;利用定向克隆技术将A和B 基因插入到载体自杀质粒pPHU281中,并添加大观霉素抗性基因片段,重组的pPHU281 _A_Spec_B 在大肠杆菌DH-5α内扩增后酶切电泳鉴定并进行DNA测序.结果 通过对重组质粒pPHU281_A_Spec_B进行酶切鉴定以及DNA序列测定分析,证明重组质粒pPHU281_A_Spec_B构建成功.结论 成功构建了质粒pPHU281_A_Spec_B,有利于菌毛蛋白FimA缺陷型Pg的构建,为深入研究FimA的作用机制奠定了基础.     

Association of Periodontal Diseases and Liver Fibrosis in Patients With HCV and/or HBV infection

Background:

Periodontal disease and systemic health are closely associated. However, there is no data supporting the association between periodontal disease and patients with liver diseases associated with hepatitis C virus (HCV) and/or hepatitis B virus (HBV) infection.

Objectives:

The aim of this study was to evaluate the association between periodontitis and progression of liver diseases in patients with HCV and/or HBV infection.

Patients and Methods:

In this retrospective study, 351 patients with HCV- and/or HBV-related liver diseases underwent screening for periodontal disease using the Salivaster® salivary occult blood test from February 2010 to June 2014. Furthermore, we examined the prevalence of fimbrillin (fimA) genotype of Porphyromonas gingivalis (P. gingivalis) in 28 HCV-infected patients visited at our hospital between January 2013 and June 2014. P. gingivalis with fimA genotype with types I to V was further detected using a PCR method.

Results:

Of 351 patients, 76 patients (group 1) had a strong positive result for salivary occult blood test and 275 patients (group 2) had weak positive or negative test results. Significant factors between the groups were obesity, level of AST, ALT, LDH, ALP, Alb, D.Bil, T.cho, AFP, platelets (Plt), IRI, HOMA-IR, current interferon (IFN) treatment and the daily frequency of tooth brushing. Between-groups analysis indicated that total protein (T.pro) level and liver fibrosis were significant factors. According to multivariate analysis, five factors were associated with periodontal disease as Plt count below 80000, brushing teeth only once a day, current IFN treatment, aged 65 years or older and obesity. The adjusted odds ratios for these five factors were 5.80, 3.46, 2.87, 2.50 and 2.33, respectively, and each was statistically significant. Twenty-eight saliva specimens had positive results for P. gingivalis with fimA genotype types I to V. The prevalence of fimA genotype II was higher in 14 patients with liver cirrhosis or a history of hepatocellular carcinoma treatment (group B, 50.00%) than 14 patients with only hepatitis C (group A, 21.43%).

Conclusions:

Periodontitis might be associated with progression of viral liver disease; hence, controlling oral disease is essential for the prevention and management of liver fibrosis.     

齿龈内阿米巴的超微结构与溶酶体酶细胞化学研究

目的　研究齿龈内阿米巴的细胞器与功能 ,探讨虫体与宿主细胞的关系。方法　用透射电镜结合电镜酶细胞化学技术观察虫体细胞超微结构 ;将虫体与口腔上皮细胞孵育 ,观察虫体对上皮细胞的粘附。结果　虫体及其伪足呈多形态 ;胞质内细胞器主要为泡状和管状的溶酶体、一些滑面内质网 ,以及丰富的糖原颗粒 ;溶酶体标志酶CMPase显示溶酶体酶丰富 ;可见虫体粘附于上皮细胞表面和小伪足伸入上皮细胞微绒毛间。结论　虫体的溶酶体水解酶消化作用及虫体伪足的机械作用对宿主牙周上皮组织的损伤 ,是虫体的致病机制之一。     

Infection with a periodontal pathogen increases mononuclear cell adhesion to human aortic endothelial cells

BACKGROUND: As a link between periodontal infections and an increased risk for vascular disease has been demonstrated, we assessed the ability of the Gram-negative periodontal pathogen Porphyromonas gingivalis to modulate properties of endothelial cells linked to inflammation and proatherogenic pathways. METHODS AND RESULTS: Primary human aortic endothelial cells (HAEC) were infected with either P. gingivalis strain 381 or its non-invasive fimbriae-deficient mutant, DPG3, and incubated with U-937 monocytes, or Jurkat T cells. P. gingivalis-infected HAEC demonstrated significantly increased adhesion of immune cells compared to non-infected cells or those infected with DPG3. Heat-killed bacteria had no effect on mononuclear cell adhesion and P. gingivalis LPS had only a minimal effect. P. gingivalis infection significantly increased HAEC expression of VCAM-1, ICAM-1 and E-selectin, and enhanced production of IL-6, IL-8 and MCP-1. CONCLUSION: These data demonstrate that live invasive P. gingivalis 381 elicits a pro-atherogenic response in HAEC.     

Does a biological link exist between periodontitis and rheumatoid arthritis?

Periodontitis or Periodontal disease(PD) and Rheumatoid arthritis(RA) are two the most common chronic inflammatory diseases. Periodontitis is a biofilm associated destructive inflammatory disease of the periodontium caused by specific microorganisms. Rheumatoid arthritis is an autoimmune condition and is identified by elevated serum autoantibody titre directed against citrullinated peptides or rheumatoid factor. Periodontitis may involve some elements of autoimmunity. Recent studies have established that PD and RA show a common pathway and could be closely associated through a common dysregulation and dysfunction in inflammatory mechanism. The enzyme peptidyl arginine deiminase(PAD), expressed by Porphyromonas gingivalis(P. gingivalis) is responsible for the enzymatic deimination of arginine residuals to citrulline resulting in protein citrullination and its increased accumulation in RA.Citrullination by PAD may act as a putative biologic link between PD and RA. Association of Human leukocytic antigen-DR4 antigen has been established both with RA and PD. Several interleukins and inflammatory mediators(ILs) and Nuclear factor kappa beta ligand are linked to these common chronic inflammatory diseases. Antibodies directed against heat shock protein(hsp 70 ab) of P. gingivalis, P. melanogenicus and P. intermedia are raised in PD as well as RA. Both the conditions share many pathological and immunological similarities. Bacterial infection, genetic susceptibility, altered immune reaction and inflammatory mediators considered responsible for RA are also associated with PD. So it is plausible that a biological link may exist between PD and RA. Therapies aimed at modifying the expression and effect of inflammatory mediators and effector molecules such as matrix metalloproteinases, proinflammatory cytokines and autoantibodies of structural proteins may probably reduce the severity of both RA and PD.     

Toll样受体4介导牙龈卟啉单胞菌感染血管内皮细胞过程中的炎性作用

目的探讨Toll样受体4(Toll-like receptor 4,TLR4)在牙龈卟啉单胞菌感染血管内皮细胞过程中所发挥的作用。方法用牙龈卟啉单胞菌分别感染正常EAhy926细胞和TLR4基因敲减的EAhy926细胞后,用RT-PCR和ELISA方法观察2种细胞产生白细胞介素6(IL-6)和白细胞介素8(IL-8)的能力,用单核细胞黏附实验检测内皮细胞黏附单核细胞的作用。结果牙龈卟啉单胞菌能诱导2种细胞过度表达IL-6和IL-8,促进U937细胞的黏附(P<0.05);敲减了TLR4基因的EAhy926细胞受到牙龈卟啉单胞菌感染后,IL-6和IL8的产生以及单核细胞黏附作用明显低于正常的EAhy926细胞(P<0.05)。结论 TLR4在牙龈卟啉单胞菌引起EAhy926细胞炎性反应过程中具有重要作用。