Role of the TLR-pp65-integrin system in host defense against bacterial infections

TLR-pp65-整合素系统在宿主防御细菌感染中的作用

• 批准号: 15590390
• 负责人: SHINOMIYA Hiroto
• 金额: $ 2.3万
• 依托单位: Ehime 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-15590390/
• 关键词: p65; L-plastin; Toll-like receptor; integrin; actin; cytoskeleton; cell adhesion; host dfense; macrophage

We previously identified p65/L-plastin that was phosphorylated in LPS-stimulated macrophages. It has been clarified that the protein has Ca-binding, calmodulin-binding and actin-binding domains., and that it regulates irtegrin-mediated cell adhesion. The C3H/HeJ mouse is known to be unresponsive to LPS due to the mutation of its TLR4. The p65/L-plastin-regulated adhesion is not enhanced in LPS-stimulated macrophages of the mouse, which may be the cause of the susceptibility of the mouse to bacterial infections. In order to investigate an important role of the TLR-p65 phosphorylation-cell locomotion cascade in host defense against infections, we have carried out the following experiments.(1)TLR KO mice : TLR4,TLR2,TLR9 and MyD88 KO mice were bled and used for experiments. (2)Proteins : Recombinant protein of p65/L-plastin was prepared. Monoclonal antibody to p65/L-plastin was also prepared. In order to assess the specificity of the mAb, recombinant T-plastin that is an isoform of a plas … More tin protein family and expressed in many other tissue except leukocytes was prepared. Recombinant form of grancalcin that is supposed to form a complex with p65/L-plastin in cells was also prepared. (3)In vitro infection experiments : Macrophages and dendritic cells from various mice were stimulated in vitro with bacteria or bacterial components. The relationship between intracellular localization of p65/L-plastin or grancalcin that was immunohistochemically detected with specific antibodies and the increase in cell adhesion was assessed. (4)In vivo infection experiments : Various mice were in vivo infected with Salmonella organisms and leukocytes recruited into the infected site were obtained. It was found that intracellular arrangemert of p65/L-plastin was greatly changed in the cells, and that grancalcin was translocated to actin-cytoskeleton including p65/L-plastin. These observations strongly suggest that TLR-pp65-integrin system plays a vital role in host defense against bacterial infections. Less

项目成果

Preparation and characterization of recombinant p65/L-plastin expressed in E. coil and high-titer antibodies against the protein

大肠杆菌中表达的重组 p65/L-plastin 的制备和表征以及针对该蛋白的高滴度抗体

• 发表时间: 2003
• 期刊: Biosci.Biotech.Bichem. 67 (6)
• 作者: H.Shinomiya;et al.
• 通讯作者: et al.

Liu Fengzhi, et al.: "Characterization of murine grancalcin specifically expressed in leukocytes and its possible role in host defense against bacterial infection"Biosci.Biotech.Biochem.. (in press). (2004)

Liu Fengzhi 等人：“白细胞中特异性表达的小鼠大钙蛋白的特征及其在宿主防御细菌感染中的可能作用”Biosci.Biotech.Biochem..（出版中）。

Preparation and characterization of recombinant murine p65/L-plastin expressed in E.coli and high-titer antibodies against the protein.

在大肠杆菌中表达的重组鼠 p65/L-plastin 的制备和表征以及针对该蛋白质的高滴度抗体。

• 发表时间: 2003
• 期刊: Biosci.Biotechnol.Biochem. 67
• 作者: Shinomiya T;et al.
• 通讯作者: et al.

Characterization of murine grancalcin specifically expressed in leukocytes and its possible role in host defense against bacterial infection

• DOI: 10.1271/bbb.68.894
• 发表时间: 2004-04-01
• 期刊: BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY
• 影响因子: 1.6
• 作者: Liu, FZ;Shinomiya, H;Asano, Y
• 通讯作者: Asano, Y

Shinomiya, H., et al.: "Preparation and characterization of recombinant murine p65/L-plastin expressed in E.coli and high-titer antibodies against the protein"Biosci.Biotech.Biochem.. 67(6). 1368-1375 (2003)

Shinomiya, H. 等人：“在大肠杆菌中表达的重组鼠 p65/L-plastin 和针对该蛋白质的高滴度抗体的制备和表征”Biosci.Biotech.Biochem.. 67(6)。