CIK细胞在免疫缺陷NPG小鼠体内重复给药毒性研究  

Toxicity Study of Cytokine Induced Killer Cells in Immunodeficient NPG Mice

作　　者：黄瑛[1] 刘静维 侯田田 文海若 秦超 卢戌 耿兴超[1] HUANG Ying;LIU Jingwei;HOU Tiantian;WEN Hairuo;QIN Chao;LU Xu;GENG Xingchao(The Beijing Key Lab for Pre-clinical Safety Evaluation of Drugs,National Center for Safety Evaluation of Drugs,National Institutes for Food and Drug Control,Beijing 100176,China;Karh Biohealthcare Biotechnology(Zhejiang)Co.,Ltd.,Jiaxing 314100,China)

机构地区：[1]中国食品药品检定研究院,国家药物安全评价监测中心,药物非临床安全评价研究北京市重点实验室,北京100176 [2]康爱瑞浩生物医药(浙江)股份有限公司,浙江嘉兴314100

出　　处：《中国药学杂志》2024年第20期1899-1909,共11页Chinese Pharmaceutical Journal

基　　金：国家重点研发计划课题资助(2021YFA1101602);中国食品药品检定研究院关键技术研究基金资助(GJJS-2022-6-1)。

摘　　要：目的采用重度免疫缺陷NPG小鼠模型对细胞因子诱导的杀伤性(CIK)细胞治疗产品进行毒性研究,考察多次给药后的毒性反应,为临床应用提供安全性依据。方法采用NPG小鼠,设主实验组和卫星组两个大组,每大组分为溶媒对照组、低剂量组(每只小鼠给予2×10^(6)个细胞)和高剂量组(每只小鼠给予3×10^(7)个细胞)。每两周给药1次,共给药6次,恢复期28 d。实验期间,主实验组动物进行临床症状观察、体质量和摄食量测定,并于给药期结束和恢复期结束后解剖采血,进行血液学和血清生化检查、细胞因子检测、脏器称重和病理学检查。卫星组动物于首次给药和末次给药前、给药后3 h、1、3、10 d以及恢复期结束后采血,进行细胞表型分析,以考察CIK细胞的分布情况。结果在低剂量组重复给药6次,小鼠未见明显毒性反应,仅个别动物出现弓背、摄食量下降和葡萄糖(GLU)、总胆固醇(CHO)升高等症状,血液中仅在首次给药后1 d和末次给药后个别时间点(3 h、10 d、29 d)检测到少量CIK细胞。高剂量组重复给药6次,小鼠血液中可检测较高水平的CIK细胞,血清中γ-干扰素(IFN-γ)、肿瘤坏死因子(TNF)、白细胞介素10(IL-10),白细胞介素2(IL-2)等具有肿瘤杀伤作用的细胞因子升高,同时小鼠出现明显的移植物抗宿主反应(graft-versus-host disease,GvHD),表现为动物临床症状、体质量、摄食量、血液学和血清生化等多个指标的变化、动物多脏器发生混合细胞聚集和GvHD相关的病变。结论NPG小鼠重复给予CIK细胞,在每只小鼠给予2×10^(6)个细胞的剂量(临床拟用剂量)下动物未见显著的毒性反应;在每只小鼠给予3×10^(7)个细胞剂量下动物出现与异种细胞移植相关的GvHD反应,未见其他相关的毒理学反应。该结果为CIK细胞进入临床试验奠定了基础。OBJECTVIE To provide safety basis for clinical application,and evaluate the toxicity CIK cells by repeated administration of CIK cells in severe immunodeficient NPG mouse model.METHODS The NPG mice were randomized into two groups:the main experimental group and the satellite group.Each group was divided into vehicle control group,low dose group(2×10^(6)cells per mouse)and high dose group(3×10^(7)cells per mouse).The CIK cells were given every two weeks for 6 times with 28-day recovery.During the experiment,the clinical symptoms,body weight and food intake were observed in the main experimental group.Blood samples were dissected and collected at the end of the administration period and recovery period,and the hematology,serum biochemical examination,cytokine detection,organ weighing and pathological examination were performed.The blood samples of satellite animals were collected to investigate the distribution of CIK cells before the first and last administration,3 h,1 d,3 d,10 d after the administration,and after the recovery period.RESULTS After repeated administration for 6 times at low dose,no obvious toxic reaction was observed in mice.A few CIK cells were detected in the blood at 1 day after the first dose and at some time points(3 h,day 10,day 29)after the last dose.High levels of CIK cells were detected in the blood of mice after repeated administration of 6 times at high dose.The levels of IFN-γ,TNF,IL-10,IL-2 and Il-10 in serum were increased.At the same time,significant graft-versus-host disease(GvHD)reaction was observed in mice,including the changes of clinical symptoms,body weight,food intake,hematology and serum biochemistry,mixed cell aggregation and GvHD-related lesions occurred in multiple organs of animals.CONCLUSION The data from the study indicates that no significant toxic reaction was observed at the dose of 2×10^(6)cells per mouse(the clinical dosage).GvHD associated with xenotransplantation was observed at a dose of 3×10^(7)cells per mouse,and no other related toxicity was observed.Th

关 键 词：细胞治疗 细胞因子诱导的杀伤性细胞 免疫缺陷小鼠 安全性 

分 类 号：R96[医药卫生—药理学]