Establishment of Transformation System on Periodontopathic Bacteria and Analysis of Pathogenic Factors

牙周病菌转化体系的建立及致病因素分析

• 批准号: 11470383
• 负责人: UMEMOTO Toshio
• 金额: $ 8.13万
• 依托单位: Kanagawa 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: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-11470383/
• 关键词: Porphyromonas gingivalis; Periodontopathic bacteria; major fimbriae; minor fimbriae; inactivated fimA gene; fimA transformed strain; cell attachment; bone resorption activity; メジャーフィンブリエ; マイナーフィンブリエ; 細胞付着能; mfal不活化; fimA不活化株; mfa1不活化株; 線毛

We previously reported that the presence of two different kinds of fimbriae expressed on the cell surface of Porphyromonas gingivalis ATCC 33277. The initial event in most infectious diseases involves adhesion of pathogens to host tissues and subsequent invasion by the pathogens. To define the role of fimbriae in Porphyromonas gingivalis adherence to and invasion of epithelial cells, we have constructed fimbrial mutants. The involvement of P. gingivalis fimbriae in the invasion process and alveolar bone resorption in rats were examined.Inactivated mutants of 41-K fimbrillin gene (fimA) and/or the 67-K fimbrillin gene (mfal) by a homologous recombination technique and compared among fimA mutant (MPG1), mfal mutant (MPG67), and double knockout mutant (MPG4167). Adherence and invasion of P. gingivalis was assessed in human oral epithelial KB cells. We used a rat model to examine the role of each type of fimbriae in alveolar bone loss by oral infection.The adherence and invasion levels of the mutants were lower than the wild-type strain. The bone loss of rats infected with the MPG1 was higher than that of those infected with MPG67. Moreover, the bone loss of rats infected with the double knockout mutant was significantly decreased compared to that of rats infected with the wild-type strain.Data from this study suggest that not only the 41-K fimbrial protein, but also the 67-K fimbrial protein play important roles in the pathogenesis of periodontal disease.

我们之前报道过牙龈卟啉单胞菌 ATCC 33277 细胞表面存在两种不同的菌毛表达。大多数传染病的初始事件涉及病原体粘附到宿主组织以及随后病原体的入侵。为了确定菌毛在牙龈卟啉单胞菌粘附和侵袭上皮细胞中的作用，我们构建了菌毛突变体。检测牙龈卟啉单胞菌菌毛参与大鼠侵袭过程和牙槽骨吸收。通过同源重组技术对41-K fimbrillin 基因（fimA）和/或67-K fimbrillin 基因（mfal）灭活突变体进行比较fimA突变体(MPG1)、mfal突变体(MPG67)和双敲除突变体(MPG4167)。在人口腔上皮 KB 细胞中评估牙龈卟啉单胞菌的粘附和侵袭。我们使用大鼠模型来研究每种类型的菌毛在口腔感染引起的牙槽骨丢失中的作用。突变株的粘附和侵袭水平低于野生型菌株。感染MPG1的大鼠的骨丢失量高于感染MPG67的大鼠。此外，与感染野生型菌株的大鼠相比，感染双敲除突变体的大鼠的骨丢失显着减少。本研究的数据表明，不仅41-K菌毛蛋白，而且67-K菌毛蛋白也明显减少。菌毛蛋白在牙周病的发病机制中发挥重要作用。

项目成果

Hiroko Hiramine, Kiyoko Watanabe, Nobushiro Hamada, Toshio Umemoto.: "Induction of IL-la production and osteoclast differentiation by gingivalis minor fimbriae."The Bulletin of Kanagawa Dental of College. 31(1). 65-67 (2003)

Hiroko Hiramine、Kiyoko Watanabe、Nobushiro Hamada、Toshio Umemoto.：“牙龈小菌毛诱导 IL-la 产生和破骨细胞分化。”神奈川牙科学院通报。

Aruwa M, Hamada N, Arai M.: "Characterization of fimA mutant from Porphyromonas gingivalis 381"Kanagawa shigaku. 36. 95-103 (2001)

Aruwa M、Hamada N、Arai M.：“牙龈卟啉单胞菌 381 的 fimA 突变体的表征”神奈川志学。

Takahashi Y, Kato D, Hamada N, Yoshimoto H, Umemoto T.: "Transformation and expression of a cloned fimA gene in Porphyromonas gingivalis"Infect.Immun.. 67(4). 2133-2018 (1999)

Takahashi Y、Kato D、Hamada N、Yoshimoto H、Umemoto T.：“牙龈卟啉单胞菌中克隆 fimA 基因的转化和表达”Infect.Immun.. 67(4)。

Takahashi Y, Kato D, Hamada N, Yoshimoto H, Umemoto T.: "Transformation and expression of a cloned fimA gene in Porphyromonas gingivalis."Infect.Immun.. 67(4). 2013-2018 (1999)

Takahashi Y、Kato D、Hamada N、Yoshimoto H、Umemoto T.：“牙龈卟啉单胞菌中克隆 fimA 基因的转化和表达。”Infect.Immun.. 67(4)。

Takahashi Y, Yoshimoto H, Kato D, Hamada N, Arai M, Umemoto T.: "Reduced fimbria-associated activities of Porphyromonas gingivalis induced by recombinant fimbrial expression."FEMS Microbiology Letters. 195. 217-222 (2001)

Takahashi Y、Yoshimoto H、Kato D、Hamada N、Arai M、Umemoto T.：“重组菌毛表达诱导的牙龈卟啉单胞菌菌毛相关活性降低。”FEMS 微生物学快报。