1995 Fiscal Year Final Research Report Summary
Identification and biological properties of bacterial endotoxin
| Project/Area Number |
06670302
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
Bacteriology (including Mycology)
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| Research Institution | Jichi Medical School |
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Principal Investigator |
KIRIKAE Teruo Faculty of Medicine, Jichi Medical School Assistant, 医学部, 講師 (50192563)
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| Project Period (FY) |
1994 – 1995
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| Keywords | endotoxin / LPS / CD14 / macrophage / taxol / lipid A |
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| Research Abstract |
Biological and biochemical prpperties of bacterial endotoxin (lipopolysaccharide, LPS) were examined.
1. LPS-responsiveness of a CD14-negative cell line.
LPS-responsiveness of non-macrophage ST2 cells derived from murine bone marrow stroma was examined. ST2 cells did not express CD14 mRNA.The cells as well as macrophages expressed IL-6 mRNA in response to LPS (at a minimal does of 0.1 ng/ml), but did neither TNF nor nitric oxide. Taxol, an LPS agonist, induced IL-6 mRNA expression by ST2 cells. Phodobacter sphaeroides lipid A (RsDPLA), an LPS antagonist, inhibited both LPS-and taxol-induced IL-6 mRNA expression.
2. LPS-responsiveness and LPS-binding properties of ST2 cells transfected with murine CD14 cDNA.
To examine whether other LPS receptor molecules than CD14 are required for CD14-dependent cell activation in macrophages, we prepared LPS-responsive ST2 and LPS-non-responsive COS cells transfected with murine CD14 cDNA expression vectors.
3. CD14-independent LPS-binding in LPS-responsive cells.
To clarify whether CD14-negative ST2 cells have LPS-binding sites on the cells, binding studies of radiolabeled LPS to the cells were examined. ^<125>I-LPS showed time-and dose-saturable specific binding to ST2 cells, suggesting that there exists a CD14-independent binding sites on ST2 cells.
Collectively, these results strongly suggests a CD14-independent signal transduction pathway for IL-6 mRNA-expression in response to LPS.
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Research Products
(11 results)
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[Publications] Kirikae, T., Lei, M.-G., Chen, T., and Morrison, D.C.: "Interactions of endotoxin with membranes and membrane receptors in K.L.Brigham (Ed.) : Endotoxin and the lungs., New York, Marcel" Dekker, INC.21-44 (1994)
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[Publications] Kirikae, F., Kirikae, T., Qureshi, N., Takayama, T., Morrison, D.C., and M.Nakano.: "CD14 is not involved in Rhodobacter spheroides diphosphoryl lipid A inhibition of tumor necrosis factor alpha and nitric oxide induction by taxol in murine macrophages." Infect.Immun.63. 486-497 (1995)
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「研究成果報告書概要(欧文)」より
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[Publications] Suda, Y., Kirikae, T., Shiyama, T., Yasukochi, T., Kirikae, F., Nakano, M., Rietschel, E.Th., Kusumoto, S.: "Macrophage activation in responce to S-form lipopolysaccharides (LPS) separated by centrifugal partition chromatography from wild-type LPS : effects of the O-polysaccharide portion of LPS." Biochem, Biophys.Res.Commun.210. 678-685 (1995)
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「研究成果報告書概要(欧文)」より
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