Susceptibility to bacterial infection by gene transfection of beta defensinsjnto oral epithelial cells

口腔上皮细胞β防御素基因转染对细菌感染的易感性

• 批准号: 12671777
• 负责人: ABIKO Yoshihiro
• 金额: $ 2.24万
• 依托单位: Health Scjences University of Hokkaido
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-12671777/
• 关键词: defensin; transfection; bacterial infection; periodontitis; 抗細菌性タンパク; 口腔上皮

βDefensins (hBD) are a group of antimicrobial peptides. We have demonstrated the decreasing expression of hBDs in KB cells widely used in vitro model of periodontitis, and that Actinobacillus actinomycetemcomitant (Aa) infected into the KB cells to induce their apoptosis. The study examined whether overexpression of hBDs in KB cells by the gene transfection made them resistance to bacterial infection. We established hBD-1-and hBD-2- overexpressed KB cell lines (KB-hBD1, KB-hBD2) by gene transfection. The mRNA and protein expressions of hBDs were confirmed by RTPCR and western blot analyses, respectively. The western blot analysis showed double bands, implying that processing occurred during production of the functional protein. DNA fragmentation assay was carried out to evaluate the infected rate. The DNA fragmentation in control KB cells increased with Aa infection in a dose dependent manner. On the other hand, no significant increase of DNA fragmentation in KB-hBD1 or KB-hBD2 with Aa infection was observed. The infection rate in KB-hBD1 or KB-hBD2 was a significantly lower than control KB. The infection rate in KB-hBD1 is a significantly higher than that in KB-hBD2. These results suggest that overexpression of hBDs in epithelial cells makes them resistance to bacterial infection.

β防御素 (hBD) 是一组抗菌肽。我们已经证明在牙周炎体外模型中广泛使用的KB细胞中hBDs表达降低，并且放线杆菌伴放线杆菌（Aa）感染到KB细胞中诱导其凋亡。该研究检查了通过基因转染在 KB 细胞中过度表达 hBD 是否使它们对细菌感染具有抵抗力。我们通过基因转染建立了hBD-1-和hBD-2-过表达的KB细胞系（KB-hBD1、KB-hBD2）。 hBDs 的 mRNA 和蛋白表达分别通过 RTPCR 和蛋白质印迹分析得到证实。蛋白质印迹分析显示双条带，这意味着在功能蛋白的生产过程中发生了加工。进行DNA片段化分析以评估感染率。对照 KB 细胞中的 DNA 片段随着 Aa 感染而增加，且呈剂量依赖性。另一方面，没有观察到Aa感染的KB-hBD1或KB-hBD2中DNA片段化的显着增加。 KB-hBD1或KB-hBD2的感染率显着低于对照KB。 KB-hBD1的感染率明显高于KB-hBD2。这些结果表明，上皮细胞中 hBD 的过度表达使其能够抵抗细菌感染。

项目成果

Liu F., Abiko Y. et al.: "Expression of inflammatory cytokines and beta-defensisn 1 mRNAs in porcine epithelial rests of malassez in vito"Med Electron Microsc. 34 (3). 174-178 (2001)

Liu F.、Abiko Y. 等人：“马拉色猪上皮剩余物体内炎症细胞因子和 β-防御素 1 mRNA 的表达”Med Electron Microsc。

Nonaka.K.: "Involvement of caspases in apoptotic cell death of murine macrophages infected with Actinobacillus actinomycetemcomitans. J.Periodont.Res. 36:40-47, 2001"J.Periodont.Res.. 36. 40-47 (2001)

Nonaka.K.：“半胱天冬酶参与感染伴放线放线杆菌的鼠巨噬细胞的凋亡细胞死亡。J.Periodont.Res.36：40-47，2001”J.Periodont.Res..36.40-47(2001)

Abiko Y, Murata M, et al.: "Immunohistochemical Localization of amelogen in human odontogenic tumors, using a polyclonal antibody against bovine amelogenin"Med Electron Microsc. 34(3). 185-189 (2001)

Abiko Y、Murata M 等人：“使用针对牛牙釉蛋白的多克隆抗体对人牙源性肿瘤中的牙釉原进行免疫组织化学定位”Med Electron Microsc。

Abiko Y,Murata M. et al.: "Immunohistochemical localization of amelogenin in human odontogenic tumors, using a polyclonal antibody against bovine amelogenin"Med Electron Microsc. 34 (3). 185-189 (2001)

Abiko Y，Murata M. 等人：“使用针对牛牙釉蛋白的多克隆抗体，对人牙源性肿瘤中的牙釉蛋白进行免疫组织化学定位”Med Electron Microsc。

Ijiri R., Abiko Y. et al.: "Pigmentated intraosseous odontogenic carcinoma of the maxilla : A pediatric case report and differential diagnosis"Human Pathology. 32 (8). 880-884 (2001)

Ijiri R.、Abiko Y. 等人：“上颌骨色素性骨内牙源性癌：儿科病例报告和鉴别诊断”人类病理学。