Akabane H
Department of Legal Medicine, Hokkaido University School of Medicine, Sapporo, Japan.
Hokkaido Igaku Zasshi. 1991 Jul;66(4):510-21.
Leukotoxin, 9: 10-epoxy-12-octadecenoic acid, was reported to exist in human burned skin and neutrophils, and to have toxic effects in experimental animals and antifungal effects against rice blast disease. Leukotoxin was regarded as a toxic and/or defensive substance in living beings. The author synthesized leukotoxin from linoleic acid with peracetic acid and purified practically by thin layer chromatography. The leukotoxin synthesized was injected into guinea pigs intravenously and caused a systemic convulsion of the animal body and cardiac arrest. The leukotoxin synthesized, on the other hand, was conjugated with bovine serum albumin (BSA) by means of the mixed anhydride technique and immunized in rabbits. After immunization of leukotoxin conjugated with BSA over 4 months, the author succeeded in producing anti-leukotoxin antiserum for the first time. According to a titration test of antiserum, sixty-folds diluted antiserum was found to bind approximately 50% of methylated leukotoxin labeled with carbon 14. And unlabeled leukotoxin was detected at least 5 ng in this radioimmunoassay by use of polyethylene glycol precipitation. This antiserum had a strong specificity to leukotoxin and no cross-reactivity to the other analogs tested. The role of leukotoxin in living creatures had not been clarified yet. Therefore both the leukotoxin synthesized by this simple procedure and the anti-leukotoxin antibodies would aid the study of the mechanism of its biological activities and its histochemical investigations.
白细胞毒素，即9,10 - 环氧 - 12 - 十八碳烯酸，据报道存在于人体烧伤皮肤和中性粒细胞中，对实验动物具有毒性作用，并对稻瘟病具有抗真菌作用。白细胞毒素被视为生物体内的一种有毒和/或防御性物质。作者用过氧乙酸从亚油酸合成了白细胞毒素，并通过薄层色谱法进行了实际纯化。将合成的白细胞毒素静脉注射到豚鼠体内，导致动物全身惊厥和心脏骤停。另一方面，通过混合酸酐技术将合成的白细胞毒素与牛血清白蛋白（BSA）偶联，并在兔体内进行免疫。在用BSA偶联的白细胞毒素免疫4个月后，作者首次成功制备了抗白细胞毒素抗血清。根据抗血清的滴定试验，发现60倍稀释的抗血清能结合约50%的用碳14标记的甲基化白细胞毒素。在该放射免疫分析中，通过聚乙二醇沉淀法至少能检测到5纳克未标记的白细胞毒素。这种抗血清对白细胞毒素具有很强的特异性，与所测试的其他类似物无交叉反应。白细胞毒素在生物体内的作用尚未阐明。因此，通过这种简单方法合成的白细胞毒素及其抗白细胞毒素抗体都将有助于研究其生物活性机制及其组织化学研究。
