Role of the TLR-pp65-integrin system in host defense against bacterial infections
| Project/Area Number |
15590390
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Bacteriology (including Mycology)
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| Research Institution | Ehime University |
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Principal Investigator |
SHINOMIYA Hiroto Ehime University, School of Medicine, Associate Professor, 医学部, 助教授 (80162618)
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| Co-Investigator(Kenkyū-buntansha) |
ASANO Yoshihiro Ehime University, School of Medicine, Professor, 医学部, 教授 (70114353)
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| Project Period (FY) |
2003 – 2004
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| Project Status |
Completed (Fiscal Year 2004)
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| Budget Amount *help |
¥3,600,000 (Direct Cost: ¥3,600,000)
Fiscal Year 2004: ¥1,400,000 (Direct Cost: ¥1,400,000)
Fiscal Year 2003: ¥2,200,000 (Direct Cost: ¥2,200,000)
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| Keywords | p65 / L-plastin / Toll-like receptor / integrin / actin / cytoskeleton / cell adhesion / host dfense / macrophage |
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| Research Abstract |
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
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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
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Report
(3 results)
Research Products
(10 results)
