Studies on Endotoxin-Elicited Actication Mechanism of Macrophages Based on Tissue Factor Generation

建立灵敏、定量的组织因子偶联氨解分析方法。凝血酶的形成已建立。一种能产生荧光的多肽底物。BOC-Val-Pro-Arg-MCA。被发现适合于偶联氨解分析。采用酰胺法测定内毒素刺激的单核/巨噬细胞的转铁蛋白活性，用脂多糖和单核细胞共培养，激活猕猴单核细胞产生组织因子。当浓度为10^~(-5)~10^~(-1)g/ml时，激活作用呈剂量依赖关系。脂多糖分子的一部分对于诱导单核细胞产生组织因子的能力是必不可少的。因此，脂多糖和细胞之间的脂-脂相互作用是激发这种能力的可能触发因素。当生理淋巴细胞与单核细胞比例为8：1~10：1时，淋巴细胞可使单核细胞产生因子的基本能力提高一倍。这表明淋巴细胞对内毒素介导的单核细胞能力具有互补作用。内毒素刺激的淋巴细胞的培养上清液影响单核细胞的活性，而刺激的淋巴细胞不影响单核细胞的活性。这一结果表明淋巴细胞的一种可溶产物。I__-.E__-。在脂多糖激活的单核细胞中，参与组织因子产生的不是细胞实体，而是类淋巴因子介质。

A sensitive and quantitative amidolytic assay for tissue factor coupled to. Thrombin formation was established. A fluogenic peptide substrate. Boc-Val-Pro-Arg-MCA. was found to be suitable for the coupled amidolytic assay. The amidolytic assay was applied to measure TF activity of endotoxin-stimulated monocytes/macrophages.Monkey (Macaca fuscata)monocytes were activated to produce tissue factor upon coculture with lipopolysaccharide, LPS. The activation was dose-related in the concentration of 10^<-5> to 10^<-1> g/ml of LPS. Lipid A portion of the LPS molecule was essential to induce the monocytes ability for tissue factor generation. Thus, a lipid-lipid interaction between LPS and the cell is a plausible trigger for eliciting the ability. Lymphocytes amlified the basal ability of monocytes to produce the factor by two-fold at physiological lymphocyte-monocyte ratios of 8:1 to 10:1. This indicates a complementary effect of lymphocyte upon the LPS-mediated monocyte ability. The medium supernatant from LPS-stimulated lymphocytes affected the monocyte competence while the stimulated lymphocytes did not. This result suggests that a soluble product of lymphocytes. i__-.e__-. lymphokine-like mediator, But not the cellular entity, participates in tissue factor production in the LPS activated monocytes.