Gene profiling of periodontal pathogens in periodontal lesion

牙周病变中牙周病原体的基因谱分析

• 批准号: 15592187
• 负责人: ARAI Hideo
• 金额: $ 2.11万
• 依托单位: OKAYAMA UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2003
• 资助国家: 日本
• 起止时间: 2003 至 2004
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-15592187/
• 关键词: periodontal bacteria; periodontitis; A.actinomycetemcomitans; host-induced genes; virulence factors; LAMP; 付着; 定着; 細胞侵入性; サブトラクトハイブリダイゼーション; 歯周病

1.Host-induced genes of A.actinomycetemcomitansActinobacillus actinomycetemcomitans possess many virulence factors. However, the potential roles of the virulence factors are not well characterized. In addition, many unknown virulence factors are considered to be involved in the pathogenesis. A.actinomycetemcomitans grows as a rough colony on primary isolates (R-type). The colony converts to a smooth phenotype (S-type) through repeated subculture. Since the phenotypic alteration reflects different gene expressions in response to environmental changes from the in vivo to the in vitro, isolation of genes induced in the R-type strain turns out to be an isolation of host-induced genes, including the virulence factors. In the current study, we identified genes induced in the R-type of A.actinomycetemcomitans using the cDNA subtractive hybridization technique.Three genes, mip,prx and ompA were identified as R-type specific genes. Attention was focused on the mip, and a recombinant Mip-like protein and the deficient mutant were created. The recombinant protein reacted with the patients sera, suggesting the production of the Mip-like protein in periodontal lesions. The mutant was used for an invasion assay and demonstrated impaired ability for the invasion of the host cells. The expression of the mip, prx and ompA may be enhanced in the host, and the Mip may play an important role for invasion of A.actinomycetemcomitans.2.New system for microbiological examination.Microflora of subgingival plaque is a very complicated community. Therefore, microbiological examination is required for the analysis of periodontal pathogens and virulence factors in vivo. In the current study, we established a new system for the examination. Loop-mediated isothermal amplification (LAMP) method was employed for the system. Primers were designed from the nucleotide sequence of 16S ribosomal RNA gene, and the LAMP method detected periodontal pathogens with high sensitivity, specificity and rapidity.

1.伴放线放线杆菌的寄主诱导基因伴放线放线杆菌具有多种毒力因子。然而，毒力因子的潜在作用还没有得到很好的表征。此外，许多未知的毒力因子被认为参与了致病过程。伴放线菌在初级分离株（R型）上生长为粗糙菌落。通过反复传代培养，殖民地转化为光滑表型（S型）。由于表型改变反映了从体内到体外响应环境变化的不同基因表达，因此在R型菌株中诱导的基因的分离结果是宿主诱导的基因的分离，包括毒力因子。本研究利用cDNA消减杂交技术对伴放线放线菌R型诱导基因进行了鉴定，发现mip、prx和ompA三个基因为R型特异基因。人们的注意力集中在mip上，并产生了重组的mip样蛋白和缺陷突变体。重组蛋白与患者血清发生反应，提示牙周病变组织中存在Mip样蛋白。将突变体用于侵袭测定，并证明其侵袭宿主细胞的能力受损。mip、prx和ompA在宿主体内的表达可能增强，Mip可能在伴放线放线菌的入侵中起重要作用。2.微生物检测新体系龈下菌斑菌群是一个非常复杂的菌群。因此，需要进行微生物检查，以分析牙周致病菌和体内毒力因子。在本次研究中，我们建立了一个新的考试体系。采用环介导等温扩增（LAMP）方法。根据16S核糖体RNA基因核苷酸序列设计引物，建立了检测牙周致病菌的LAMP方法，具有较高的敏感性、特异性和快速性。

项目成果

Detection of periodontal pathogen Porphyromonas gingivalis by loop-mediated isothermal amplification method

• DOI: 10.1016/j.femsim.2004.08.005
• 发表时间: 2005-02-01
• 期刊: FEMS IMMUNOLOGY AND MEDICAL MICROBIOLOGY
• 作者: Maeda, H;Kokeguchi, S;Takashiba, S
• 通讯作者: Takashiba, S

Periodontal treatment in severe aplastic anemia - a case report -

严重再生障碍性贫血的牙周治疗-病例报告-

• 发表时间: 2005
• 期刊: Journal of Periodontology (In press)
• 作者: 田中丈也 他;Takada K (Uematsu S as a collaborator for translation);Oyaizu K.et al.
• 通讯作者: Oyaizu K.et al.

Porphylomonas gingivalis infection contributes to elevated LDL cholesterol in diabetic subject

牙龈卟啉单胞菌感染导致糖尿病患者低密度脂蛋白胆固醇升高

• 期刊: Journal of Dental Research Special Issue (in press)
• 作者: Fusanori Nishimura;et al.
• 通讯作者: et al.