| Project/Area Number |
09470066
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Experimental pathology
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| Research Institution | Oita Medical University |
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Principal Investigator |
YAMAMOTO Shunsuke Oita Medical University, Pathology Professor, 医学部, 教授 (90040188)
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| Co-Investigator(Kenkyū-buntansha) |
MATSUURA Keiko Oita Medical University, Pathology Assistant, 医学部, 助手 (00291542)
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| Project Period (FY) |
1997 – 1999
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| Project Status |
Completed (Fiscal Year 1999)
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| Budget Amount *help |
¥5,400,000 (Direct Cost: ¥5,400,000)
Fiscal Year 1999: ¥2,700,000 (Direct Cost: ¥2,700,000)
Fiscal Year 1998: ¥2,700,000 (Direct Cost: ¥2,700,000)
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| Keywords | CD14 / TLR / ADAM / TACE / CD156 / Ostecpontin / HUVEC / オステオポンチン / エンドトキシン / メタロプロテアーゼ / ジスインテグリン / トランスジェニック / AD56 |
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| Research Abstract |
Studies on CD14 and Toll-like receptor: CD14-transgenic mice (M14S), Which expressed high level of soluble CD 14, showed low levels of blood TNF-α and IL-6 in response to LPS, less marked Shwartzman reaction, liver damage in P. acnes/LPS model and B-16 melanoma metastasis. A high level of Toll-like receptor (TLR)4 mRNA expression was found in human umbilical vein endothelial cells (HUVEC), human pereipheral blood mononuclear cells (PBMC) and THP-1 cells. Little or no TLR2 mRNA expression was observed in HUVEC. In contrast, strong TLR2 mRNA expression was observed in PBMC and THP-1 cells. Moderate and high levels of TLR1 mRNA expression were found in HUVEC, PBMC and THP-1 cells, respectively. LPS upregulated the expression of TLR4 in HUVEC. The LPS-induced E-selectin expression in HUVEC was significantly inhibited by pretreatment with an anti-TLR4 mAB. Preincubation of HUVEC with an anti-TLR4 mAb significantly reduced the LPS-induced IL-6 production. Anti-CD14 strongly inhibited the LPS
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-induced E-selectin expression and IL-6 production by HUVEC. Studies on ADAM proteins: Anti-mouse and human ADAM17 (TACE) mAb (MaTACE) was produced. Flow cytometry analysis (FCA) showed that TACE expression was induced in PMN after LPS stimulation in vitro. Immunocytochemical examination revealed that TACE was stained in unstimulated PMN as markedly as stimulated PMN. In addition, FCA showed that intracellular TACE was found in high percentage of unstimulated human PMN as LPS-stimulated PMN. The LPS-induced TNF-α production by human PBMC and mouse resident macrophages was inhibited in the presence of MaTACE. In contrast, the LPS-Induced TNF-α production by human PMN was not inhibited in the presence of MaTACE, suggesting the involvement of other sheddases and different roles of TACE in PMN. ADAM15 (AD56) : Anti-mouse ADAM15 mAb was produced. FCA using the mAb showed that ADAM15 was expressed in CD4ィイD1+ィエD1CD8ィイD1+ィエD1 and CD4ィイD1+ィエD1CD8ィイD1-ィエD1 T cells after stimulation with concanavalin A and IL-2. ADAM8 (CD 156): FCA was performed using anti-human and mouse ADAM8 mAb (1-11-G). ADAM8 was expressed in unstimulated monocytes and PMN and upregulated after stimulation with G-CSF and GM-CSF in PMN. Recombinant ectodomain of ADAM8 proteins were produced in mammalian cells. Cloning of novel ADAM cDNAs : Two cDNAs encoding novel ADAM proteins were cloned. Studies on osteopontin (OPN): Homozygote of OPN transgenic mice was produced and the investigation on their ability in inflammation and tumor metastasis were now underway. Less
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