Integrated Research on pathogenic factors of periodontal pathogens

牙周病原菌致病因素的综合研究

• 批准号: 17591912
• 负责人: SUGAI Motoyuki
• 金额: $ 2.24万
• 依托单位: HIROSHIMA UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2005
• 资助国家: 日本
• 起止时间: 2005 至 2006
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-17591912/
• 关键词: Periodontopathogenic; Cdt; Cytolethal Distending Toxin; antimicrobial peptide; defensin; A.actinomycetemcomitans; CDT; Omp; 歯周病細菌; 細胞膨化壊死毒素

Study on A. actinomycetemcomitans Cytolethal Distending Toxin (CDT) : CDT is a protein toxin composed of three subunit, CdtA, CdtB and CdtC. A. actinommycetemcomitans CdtA posseses lipobox, consensus domain for lipid modification in its C-terminal. We demonstrated that 16^<th> cys in A. actinommycetemcomitans CdtA expressed in E. coli was modified with tritiated palmitate or glycerol. Posttranscriptional prosesing of the CdtA is inhibited by globomycin, a specific inhibitor for signal peptidase II, signal peptidase for lipoprotein. We also demonstrated that the CdtA expressed in E. coli is further processed at its N-terminal in periplasmic space and is converted to CdtA'. Furthermore, we showed that lipid-modification of CdtA is important for the secretion of Cdt complex (holotoxin) into culture media. We also demonstrated that there is a SNP in cdtB coding sequences. Hence, 281th amino acid of CdtB derived from clinically isolated A. actinomycetemcomitans is either H or R. Bioassay wi … More th mutated CdtB by site-directed mutagenesis indicated that positively charged amino acid at 281 is critical for Cdt activity.Study on A. actinomycetemcomitans outer membrane protein: We have previously demonstrated that A. actinomycetemcomitans posseses several outer membrane proteins (Omp) and Omp100 has pleiotropic biological activities. We identified Omp100 as a major bacterial component inducing production of antimicrobial peptides from human gingival epidermal cells. Inhibitor assay strongy suggested that signaling pathway for hBD2 production is not by NF-kB pathway but by MAP-kinase pathway. Omp100 induces hBD2 production through associating with fibronectin and by way of integrin alpha5betal pathway. Cytokine induced by attachment of bacteria to epidermal cells also induces hBD2 expression.Study on inducibility of antimicrobial peptides by periodontopathogenic bacteria. By using synthtic peptides, we evaluated susceptibility of periodontopathogenic bacteria to antimicrobial peptides. Our data indicated that LL37 and beta-defensin showed potent antimicrobial activity against periodontapathogenic bacteria. Less

伴放线放线菌细胞致死膨胀毒素(CDT)的研究：CDT是由CdtA、CdtB和CdtC三个亚基组成的蛋白质毒素。 A. actinomycetemcomitans CdtA 拥有 lipobox，这是其 C 端脂质修饰的共有结构域。我们证明了在大肠杆菌中表达的A. actinomycetemcomitans CdtA中的第16个半胱氨酸被氚化棕榈酸酯或甘油修饰。 CdtA 的转录后加工受到球霉素的抑制，球霉素是信号肽酶 II（脂蛋白信号肽酶）的特异性抑制剂。我们还证明了大肠杆菌中表达的 CdtA 在其 N 端在周质空间中被进一步加工并转化为 CdtA'。此外，我们发现 CdtA 的脂质修饰对于 Cdt 复合物（全毒素）分泌到培养基中很重要。我们还证明了cdtB编码序列中存在SNP。因此，来自临床分离的 A. actinomycetemcomitans 的 CdtB 第 281 个氨基酸是 H 或 R。通过定点诱变对突变的 CdtB 进行生物测定表明，第 281 位带正电荷的氨基酸对于 Cdt 活性至关重要。 A. actinomycetemcomitans 外膜蛋白的研究：我们之前已经证明 A. actinomycetemcomitans 拥有多种外膜蛋白 (Omp)，Omp100 具有多效性生物活性。我们确定 Omp100 是诱导人类牙龈表皮细胞产生抗菌肽的主要细菌成分。抑制剂测定强烈表明hBD2产生的信号传导途径不是通过NF-kB途径而是通过MAP激酶途径。 Omp100 通过与纤连蛋白结合并通过整合素 alpha5beta 途径诱导 hBD2 产生。细菌附着表皮细胞诱导的细胞因子也诱导hBD2表达。牙周病原菌诱导抗菌肽的研究。通过使用合成肽，我们评估了牙周病原菌对抗菌肽的敏感性。我们的数据表明 LL37 和 β-防御素对牙周病原细菌表现出有效的抗菌活性。较少的

项目成果

Single nucleotide polymorphism in the cytolethal distending toxin B gene confers heterogeneity in the cytotoxicity of A. actinomycetemcomitans

细胞致死膨胀毒素 B 基因中的单核苷酸多态性赋予 A. actinomycetemcomitans 细胞毒性的异质性

• 发表时间: 2006
• 期刊: Infection and Immunity 74. 12
• 作者: Yamaai T;Nakanishi T;Asano M;Nawachi K;Yoshimichi G;Ohyama K;Komori T;Sugimoto;T Takigawa M;Yoko Ueno et al.;Shuichi Nishikubo et al.
• 通讯作者: Shuichi Nishikubo et al.

Cytolethal distending toxin and its implication in periodontal diseases

细胞致死膨胀毒素及其在牙周疾病中的意义

• 发表时间: 2005
• 期刊: Journal of Oral Biosciences 47-1
• 作者: M.Ohara;M.Sugai
• 通讯作者: M.Sugai

Susceptibilities of periodontopathogenic and cariogenic bacteria to antimicrobial peptides, beta-defensins and LL37, produced by human epithelial cells.

牙周致病菌和致龋菌对人上皮细胞产生的抗菌肽、β-防御素和 LL37 的敏感性。

• 发表时间: 2005
• 期刊: Journal of Antimicrobial Chemotherapy 55
• 作者: 薗村貴弘;中村公一;日置寛之;山中淳之;植村正憲;金子武嗣;Nobuhisa T;秦 正樹;Asami S;Ozaki A;薗村貴弘;Kazuhisa Ouhara et al.
• 通讯作者: Kazuhisa Ouhara et al.

Single nucleotide polymorphism in the cytolethal distending toxin B gene confers heterogeneity in the cytotoxicity of A. ctinomycetemcomitans

细胞致死膨胀毒素 B 基因中的单核苷酸多态性赋予 A. ctinomycetemcomitans 细胞毒性的异质性

• 发表时间: 2006
• 期刊: Infection and Immunity 74.12
• 作者: Kimura;S.;Yoshida K;Gunduz E;Shuichi Nishikubo et al.
• 通讯作者: Shuichi Nishikubo et al.

Biogenesis of the Actinobacillus actinomycetemcomitans cytolethal distending toxin.

伴放线放线杆菌细胞致死膨胀毒素的生物发生。

• 发表时间: 2006
• 期刊: Infection and Imunity 74(6)
• 作者: Kimura;S.;Naomoto Y;Gunduz E;Yoko Ueno et al.
• 通讯作者: Yoko Ueno et al.