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

一种灵敏、定量的组织因子偶联酶解试验。建立凝血酶形成。一种荧光肽底物。Boc-Val-Pro-Arg-MCA。被发现适合于偶联胺解试验。采用酶解法测定内毒素刺激单核/巨噬细胞的TF活性。猴（Macaca fuscata）单核细胞与脂多糖（LPS）共培养后被激活产生组织因子。在10^<-5> g/ml至10^<-1> g/ml LPS浓度范围内，激活呈剂量相关。脂质脂多糖分子的一部分是诱导单核细胞产生组织因子的能力所必需的。因此，脂多糖和细胞之间的脂质相互作用是引发这种能力的一个合理的触发器。当淋巴细胞与单核细胞的生理比例为8:1至10:1时，淋巴细胞将单核细胞产生该因子的基本能力提高了两倍。这表明淋巴细胞对lps介导的单核细胞能力有互补作用。lps刺激淋巴细胞的培养基上清影响单核细胞的能力，而刺激淋巴细胞不影响单核细胞的能力。这一结果提示淋巴细胞的可溶性产物。i__ -.e__ -。淋巴因子样介质，而非细胞实体，参与LPS活化单核细胞的组织因子生成。

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.