Biemond B J, Levi M, ten Cate H, Soule H R, Morris L D, Foster D L, Bogowitz C A, van der Poll T, Büller H R, ten Cate J W
Center for Hemostasis, Thrombosis, Atherosclerosis and Inflammation Research, University of Amsterdam, The Netherlands.
Thromb Haemost. 1995 Feb;73(2):223-30.
Gram-negative sepsis is oftentimes complicated by activation of coagulation with disseminated intravascular coagulation and microthrombosis. This may contribute to the associated morbidity, multiple organ failure and death. Recent studies have established that the tissue factor-dependent pathway of blood coagulation has a significant participatory role in the initial endotoxin-induced activation of coagulation. Tissue factor (TF), expressed on the surface of activated monocytes and endothelial cells forms cell surface complexes with free circulating factors VII and VIIa. The latter complex proteolytically activates factors X and IX. Recent in vivo experiments have shown that a rapidly neutralizing TF monoclonal antibody prevents and arrests the endotoxin-induced activation of coagulation and similar studies have shown to reduce mortality in baboons. In this study we describe the preparation of a factor VII/VIIa neutralizing monoclonal Fab fragment and characterize its effect on in vivo activation of coagulation during experimental endotoxemia in chimpanzees. Four chimpanzees received a bolus intravenous injection of 4 ng/kg endotoxin in combination with Fab fragments of a factor VII/VIIa neutralizing murine monoclonal antibody (12D10) at a dose of either 50 micrograms/kg (n = 2) or 100 micrograms/kg (n = 2). Four control animals received a bolus injection of endotoxin alone. Administration of the 12D10 Fab fragments, immediately preceding the endotoxin bolus injection, effectively blocked the endotoxin-induced activation of coagulation. Plasma levels of products of in vivo activation, namely F1 + 2, TAT complexes and FpA remained at baseline values. The administration of 12D10 resulted in a rapid decline in factor VII/VIIa antigen levels which remained below 5 ng/ml for 180-240 min, followed by a rapid return to baseline levels.(ABSTRACT TRUNCATED AT 250 WORDS)
革兰氏阴性菌败血症常常因弥散性血管内凝血和微血栓形成导致的凝血激活而变得复杂。这可能导致相关的发病率、多器官功能衰竭和死亡。最近的研究表明,凝血的组织因子依赖性途径在内毒素诱导的凝血初始激活中起重要作用。组织因子(TF)在活化的单核细胞和内皮细胞表面表达,与游离循环的因子VII和VIIa形成细胞表面复合物。后一种复合物通过蛋白水解激活因子X和IX。最近的体内实验表明,一种快速中和TF的单克隆抗体可预防和阻止内毒素诱导的凝血激活,类似研究表明这可降低狒狒的死亡率。在本研究中,我们描述了一种因子VII/VIIa中和单克隆Fab片段的制备,并表征了其对黑猩猩实验性内毒素血症期间体内凝血激活的影响。四只黑猩猩静脉注射4 ng/kg内毒素,同时注射因子VII/VIIa中和鼠单克隆抗体(12D10)的Fab片段,剂量分别为50微克/千克(n = 2)或100微克/千克(n = 2)。四只对照动物仅接受内毒素静脉推注。在内毒素推注前立即给予12D10 Fab片段,可有效阻断内毒素诱导的凝血激活。体内激活产物,即F1 + 2、TAT复合物和FpA的血浆水平保持在基线值。给予12D10导致因子VII/VIIa抗原水平迅速下降,在180 - 240分钟内保持低于5 ng/ml,随后迅速恢复到基线水平。(摘要截短于250字)