Chino F
Department of Safety Research on Biologics, National Institute of Health, Tokyo, Japan.
Dev Biol Stand. 1996;86:53-62.
NIH Japan has tested and regulated the three Rs of animal experiments in the development and control of biological products in a stepwise manner. (1) The number of monkeys was reduced from 108 to 72 for the neurovirulence test of OPV in each type, since paralysed monkeys inoculated intraspinally revealed a linear relationship between average scores of the lumbar lesion and cumulated paralysis occurrence ratio (%). (2) Rabbits for the pyrogen test were replaced by the endotoxin test for PPF, albumin and interferon products. The endotoxin is measured by the parallel line assay method using both turbidimetric kinetic and colorimetric methods. (3) Histopathological examination was introduced to the abnormal toxicity test as a refinement. Mean body weight loss of two guinea pigs inoculated with five ml. of an albumin product in each was far below the mean weight of pooled guinea pigs used (P < or = 0.01) and appeared repeatedly. The histopathological examination showed focal necrosis in the liver. This finding was suggestive of the presence of endotoxin in the product. The product contained 0.1 EU/ml of endotoxin. The same amount of the reference endotoxin produced a similar change in guinea pigs. The mechanism of the liver cell damage by endotoxin has been investigated by an in vitro method.
日本国立卫生研究院（NIH Japan）已逐步在生物制品的研发和控制过程中对动物实验的“3R”原则进行了测试和规范。（1）口服脊髓灰质炎疫苗（OPV）每种类型的神经毒力测试中，猴子数量从108只减少至72只，因为脊髓内接种的麻痹猴子显示，腰椎损伤平均评分与累积麻痹发生率（%）之间呈线性关系。（2）对于人血丙种球蛋白（PPF）、白蛋白和干扰素产品的热原试验，用内毒素试验取代了兔子热原试验。内毒素采用比浊动力学法和比色法的平行线测定法进行测量。（3）作为一种优化措施，组织病理学检查被引入异常毒性试验。每只接种5毫升白蛋白产品的两只豚鼠的平均体重减轻远低于所用合并豚鼠的平均体重（P≤0.01），且反复出现。组织病理学检查显示肝脏有局灶性坏死。这一发现提示该产品中存在内毒素。该产品含有0.1 EU/ml的内毒素。相同量的参考内毒素在豚鼠身上产生了类似变化。已通过体外方法对内毒素导致肝细胞损伤的机制进行了研究。
