Abstract: In order to evaluate the safety of recombinant human rhinovirus 3C fusion protease (GST-3C), the toxicity of GST-3C was studied by single-dose toxicity test and immunotoxicity test. Single-dose toxicity test: a drug administration group was set up, and 10 male and 10 female ICR mice were prepared. GST-3C was injected into the tail vein by maximum dose method, and the maximum dose of single dose was 25.25 mg/kg. Another normal saline control group was set up. The symptoms, degree, nature, recovery, and death of ICR mice were observed, and the mice were dissected in time at the end of the observation period to observe the effects of GST-3C on the organs of mice. Immunotoxicity test: 50 mice were randomly divided into five groups (solvent control, cyclophosphamide positive control group, adjuvanted GST-3C low-dose group, adjuvanted GST-3C high-dose group, GST-3C control group without adjuvant). Spleen index, thymus, index and swelling degree were calculated by delayed hypersensitivity method to observe the effect of GST-3C on delayed hypersensitivity. 40 mice were randomly divided into four groups (solvent control, adjuvanted GST-3C low-dose group, adjuvanted GST-3C high-dose group, GST-3C control group without adjuvant). The effect of hemolytic plaque on humoral immune function of mice was evaluated by counting hemolytic plaque test. Single-dose toxicity test showed that compared with the normal saline control group, there was no significant difference in the body weight of the mice in the experimental group, and no abnormal performance was found in the organs of the animals in the administration group. Under the conditions of this experiment, the maximum tolerated dose (MTD) of a single intravenous injection of GST-3C in ICR mice was greater than 25.25 mg/kg. Immunotoxicity test showed that during the administration period, compared with the solvent control group, the body weight of the animals in each group administered with GST-3C did not decrease significantly, and no abnormal changes related to administration were observed beside the animal cage. The results of delayed hypersensitivity test and hemolytic plaque test showed that GST-3C had no inhibitory effect on the cellular immunity and humoral immunity of animals, which provides a basis for evaluating the safety of GST-3C.

Key words: Recombinant human rhinovirus 3C fusion protease, Single-dose toxicity test, Immunotoxicity test, Safety

摘要： 为了评价重组人鼻病毒3C融合蛋白酶(GST-3C)的安全性，本工作通过单次给药毒性试验和免疫毒性试验对GST-3C的毒性进行了研究。设置一个给药组，准备雌雄ICR小鼠各10只，采用最大给药量法，分别尾静脉注射GST-3C，单次给药最大给药剂量为25.25 mg/kg；实验另设一个生理盐水对照组。给药一次，观察ICR小鼠的中毒症状、中毒程度、性质、恢复情况和死亡等，并在观察期结束对动物及时进行解剖，观察GST-3C对小鼠脏器的影响。对免疫毒性进行试验，50只小鼠随机分为5组(溶剂对照组、环磷酰胺阳性对照组、含佐剂GST-3C低剂量组、含佐剂GST-3C高剂量组和不含佐剂GST-3C对照组)，通过迟发型超敏反应方法，计算脾指数、胸腺指数和肿胀度，观察GST-3C对迟发超敏反应的影响；40只小鼠随机分为4组(溶剂对照组、含佐剂GST-3C低剂量组、含佐剂GST-3C高剂量组和不含佐剂GST-3C对照组)，通过溶血空斑试验方法，对溶血空斑计数，评价其对小鼠体液免疫功能影响。单次给药毒性试验结果表明，和生理盐水对照组相比，实验组小鼠体重差异并不明显，给药组动物脏器均未发现异常表现情况。在本次试验条件下，ICR小鼠静脉注射GST-3C单次给药的最大耐受剂量(MTD)大于25.25 mg/kg。免疫毒性试验结果表明，给药期间，与溶剂对照组比较，GST-3C给药各组动物体重未见显著降低，动物笼旁观察未见与给药相关的异常变化，迟发超敏试验和溶血空斑试验结果分别显示GST-3C对动物的细胞免疫功能和体液免疫功能均无抑制作用，为评价GST-3C的安全性提供了依据。

关键词: 重组人鼻病毒3C融合蛋白酶, 单次给药毒性试验, 免疫毒性试验, 安全性