Molecular biological analysis of the exotoxin derived from periodontopathic bacteria on peridontal medicine

牙周病菌外毒素在牙周医学中的分子生物学分析

• 批准号: 16390615
• 负责人: NISHIHARA Tatsuji
• 金额: $ 8.19万
• 依托单位: Kyushu Dental College
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2004
• 资助国家: 日本
• 起止时间: 2004 至 2005
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-16390615/
• 关键词: periodontopathic bacteria; A. ACTINOMYCETEMCOMITANS; exotoxin; CDT; monocyte-macrophage; apoptosis; glycosphingolipids; caspase; 単球・リンパ系細胞

Actinobacillus actinomycetemcomitans produces a toxin, called cytolethal distending toxin (CDT), which causes host cell DNA damage leading to the induction of DNA damage checkpoint pathways. CDT consists of three subunits, CdtA, CdtB, and CdtC CdtB is the active subunit of CDT and exerts its effect as a nuclease that damages nuclear DNA, triggering cell cycle arrest. In the present study, we confirmed that the only combination of toxin proteins causing cell cycle arrest was that of all three recombinant CDT (rCDT) protein subunits. Furthermore, in order for rCDT to demonstrate toxicity, it was necessary for CdtA and CdtC to access the cell before CdtB. The coexistence of CdtA and CdtC was necessary for these subunits to bind to the cell. Cells treated with the glucosylceramide synthesis inhibitor 1-phenyl-2-palmitoylamino-3-morpholino-1-propanol showed resistance to the cytotoxicity induced by rCDT. Furthermore, LY-B cells, which are deficient in the biosynthesis of sphingolipid, also showed resistance to the cytotoxicity induced by rCDT. To evaluate the binding of each subunit for glucosylceramides, we performed thin-layer chromatography immunostaining. The results indicated that each subunit reacted with the GM1, GM2, GM3, Gb3, and Gb4. The rCDT mixture incubated with liposomes containing GM3 displayed partially reduced toxicity. These results indicate that GM3 can act as a CDT receptor.

伴放线放线杆菌产生一种称为细胞致死性膨胀毒素（CDT）的毒素，其引起宿主细胞DNA损伤，导致DNA损伤检查点途径的诱导。CDT由三个亚基组成，CdtA、CdtB和CdtC CdtB是CDT的活性亚基，并发挥其作为核酸酶的作用，损伤核DNA，触发细胞周期停滞。在本研究中，我们证实，唯一的组合毒素蛋白引起细胞周期阻滞是所有三个重组CDT（rCDT）蛋白亚基。此外，为了证明rCDT的毒性，CdtA和CdtC必须在CdtB之前进入细胞。CdtA和CdtC的共存是这些亚基与细胞结合所必需的。用葡糖神经酰胺合成抑制剂1-苯基-2-棕榈酰氨基-3-吗啉代-1-丙醇处理的细胞显示出对rCDT诱导的细胞毒性的抗性。此外，缺乏鞘脂生物合成的LY-B细胞也显示出对rCDT诱导的细胞毒性的抗性。为了评估葡萄糖神经酰胺的每个亚基的结合，我们进行了薄层色谱免疫染色。结果表明，每个亚基与GM 1、GM 2、GM 3、Gb 3和Gb 4反应。与含有GM 3的脂质体孵育的rCDT混合物显示部分降低的毒性。这些结果表明，GM 3可以作为CDT受体。

项目成果

Antimicrobial effect of ozonated water on bacteria invading into dentinal tubules.

臭氧水对侵入牙本质小管的细菌具有抗菌作用。

• 发表时间: 2004
• 期刊: J. Endodontics 30
• 作者: Nagayoshi;M. et al.
• 通讯作者: M. et al.

Thermotolerance of pulp cells and phagocytosis of apoptotic pulp cells by surviving pulp cells following heat stress

热应激后牙髓细胞的耐热性和存活牙髓细胞对凋亡牙髓细胞的吞噬作用

• 发表时间: 2005
• 期刊: J Cell Physiol 94
• 作者: Kitamura C;Nishihara T;Ueno Y;Nagayoshi M;Kasugai S;Terashita M.
• 通讯作者: Terashita M.

Actinobacillus actinomycetemcomitans induces apoptosis in human monocytic THP-1 cells

• DOI: 10.1099/jmm.0.45693-0
• 发表时间: 2005-03-01
• 期刊: JOURNAL OF MEDICAL MICROBIOLOGY
• 影响因子: 3
• 作者: Kato, S;Sugimura, N;Kowashi, Y
• 通讯作者: Kowashi, Y

Tensile mechanical strain up-regulates Runx2 and osteogenic factor expression in human periosteal cells:Implications for distraction osteogenesis

拉伸机械应变上调人骨膜细胞中 Runx2 和成骨因子的表达：对牵引成骨的影响

• 发表时间: 2005
• 期刊: J Oral and Maxillofacial surgery 63
• 作者: Kanno T;Takahashi T;Ariyoshi W;Tsujisawa T;Iwamura M;Nishihara T.
• 通讯作者: Nishihara T.

Delivery of cytolethal distending toxin B induces cell cycle arrest and apoptosis in gingival squamous cell carcinoma in vitro.

• DOI: 10.1111/j.1600-0722.2004.00157.x
• 发表时间: 2004-10
• 期刊: European journal of oral sciences
• 影响因子: 1.9
• 作者: Kozo Yamamoto;K. Tominaga;M. Sukedai;T. Okinaga;Kenjiro Iwanaga;T. Nishihara;J. Fukuda