Identification and biological properties of bacterial endotoxin

细菌内毒素的鉴定及生物学性质

• 批准号: 06670302
• 负责人: KIRIKAE Teruo
• 金额: $ 1.34万
• 依托单位: Jichi Medical School
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1994
• 资助国家: 日本
• 起止时间: 1994 至 1995
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-06670302/
• 关键词: endotoxin; LPS; CD14; macrophage; taxol; lipid A; 細菌内毒素; リポ多糖; 受容体

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.

检测细菌内毒素（脂多糖，LPS）的生物学和生化特性。 1． CD14阴性细胞系的LPS反应性。检查了源自小鼠骨髓基质的非巨噬细胞ST2细胞的LPS反应性。 ST2 细胞不表达 CD14 mRNA。细胞和巨噬细胞响应 LPS（最低剂量为 0.1 ng/ml）表达 IL-6 mRNA，但 TNF 和一氧化氮均不表达。紫杉醇（一种 LPS 激动剂）可诱导 ST2 细胞表达 IL-6 mRNA。 LPS 拮抗剂球形磷光杆菌脂质 A (RsDPLA) 可抑制 LPS 和紫杉醇诱导的 IL-6 mRNA 表达。 2．转染鼠CD14 cDNA的ST2细胞的LPS反应性和LPS结合特性。为了检查巨噬细胞中CD14依赖性细胞活化是否需要CD14以外的其他LPS受体分子，我们制备了转染鼠CD14 cDNA表达载体的LPS反应性ST2和LPS非反应性COS细胞。 3． LPS 反应细胞中 CD14 独立的 LPS 结合。为了澄清 CD14 阴性 ST2 细胞在细胞上是否具有 LPS 结合位点，检查了放射性标记的 LPS 与细胞的结合研究。 ^ 125 I-LPS显示出与ST2细胞的时间和剂量饱和的特异性结合，表明ST2细胞上存在不依赖于CD14的结合位点。总的来说，这些结果强烈表明响应LPS的IL-6 mRNA表达存在不依赖于CD14的信号转导途径。

项目成果

KIRIKAE,F: "CD14 is not involved in Rhodobacter sphaeroides diphosphoryl lipidA inhibition of Tumor necrosisfactor alpha and nifric oxide induction by toxol in murine macroi" Infection and Immuhity. 63. 486-497 (1995)

KIRIKAE，F：“CD14 不参与球形红杆菌二磷酰脂质 A 抑制肿瘤坏死因子 α 和毒醇在小鼠宏观中诱导一氧化氮”感染和免疫。

F.KIRIKAE ら: "CD14 is notinvolved in Rhodobacter sphaeroides di phosphoryl lipidA in hibition of tumon mecrosis factor alpha and nitriconid induction." Infection and Immunity. 63. 486-497 (1995)

F.KIRIKAE 等人：“CD14 不参与球形红杆菌二磷酰脂质 A 抑制肿瘤坏死因子 α 和硝酮诱导。” 63. 486-497 (1995)。

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."

Kirikae, F.、Kirikae, T.、Qureshi, N.、Takayama, T.、Morrison, D.C. 和 M.Nakano.：“CD14 不参与球状红杆菌二磷酰脂质 A 对肿瘤坏死因子 α 和一氧化氮的抑制

Y. Sudaら: "Macrophage activition in response ToS-form lipopolysaccharidde (LPS) separated by centrifugal partitition chromatography." Biochemical and Biophysical Research Communications. 210. 678-685 (1995)

Y. Suda 等人：“通过离心分配色谱分离的 ToS 形式脂多糖 (LPS) 的巨噬细胞激活。” 210. 678-685 (1995)。

Nakano, M., Kirikae, T.: "Biological characterization of Pseudomonas aeruginosa endotoxin released by antibiotic-treatment in vitro." J.Endotoxin Res. (in press). (1996)

Nakano, M.，Kirikae, T.：“体外抗生素治疗释放的铜绿假单胞菌内毒素的生物学特征。”